CN101932711A - 晚期宫颈细胞筛查方法 - Google Patents
晚期宫颈细胞筛查方法 Download PDFInfo
- Publication number
- CN101932711A CN101932711A CN2008801093612A CN200880109361A CN101932711A CN 101932711 A CN101932711 A CN 101932711A CN 2008801093612 A CN2008801093612 A CN 2008801093612A CN 200880109361 A CN200880109361 A CN 200880109361A CN 101932711 A CN101932711 A CN 101932711A
- Authority
- CN
- China
- Prior art keywords
- leu
- thr
- val
- arg
- cys
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
Images
Classifications
-
- G01N33/5755—
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- C07K14/005—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from viruses
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K16/00—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies
- C07K16/08—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from viruses
- C07K16/081—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from viruses from DNA viruses
- C07K16/084—Papovaviridae, e.g. papillomavirus, polyomavirus, SV40, BK virus, JC virus
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N7/00—Viruses; Bacteriophages; Compositions thereof; Preparation or purification thereof
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
- G01N33/569—Immunoassay; Biospecific binding assay; Materials therefor for microorganisms, e.g. protozoa, bacteria, viruses
- G01N33/56983—Viruses
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N2710/00—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA dsDNA viruses
- C12N2710/00011—Details
- C12N2710/20011—Papillomaviridae
- C12N2710/20022—New viral proteins or individual genes, new structural or functional aspects of known viral proteins or genes
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N2333/00—Assays involving biological materials from specific organisms or of a specific nature
- G01N2333/005—Assays involving biological materials from specific organisms or of a specific nature from viruses
- G01N2333/01—DNA viruses
- G01N2333/025—Papovaviridae, e.g. papillomavirus, polyomavirus, SV40, BK virus, JC virus
Landscapes
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Organic Chemistry (AREA)
- Virology (AREA)
- Immunology (AREA)
- Engineering & Computer Science (AREA)
- Biochemistry (AREA)
- General Health & Medical Sciences (AREA)
- Genetics & Genomics (AREA)
- Medicinal Chemistry (AREA)
- Molecular Biology (AREA)
- Biomedical Technology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Zoology (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Biophysics (AREA)
- Biotechnology (AREA)
- Wood Science & Technology (AREA)
- Hematology (AREA)
- Microbiology (AREA)
- Urology & Nephrology (AREA)
- Cell Biology (AREA)
- Physics & Mathematics (AREA)
- Analytical Chemistry (AREA)
- General Physics & Mathematics (AREA)
- Pathology (AREA)
- Food Science & Technology (AREA)
- Gastroenterology & Hepatology (AREA)
- Tropical Medicine & Parasitology (AREA)
- General Engineering & Computer Science (AREA)
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
- Investigating Or Analysing Biological Materials (AREA)
- Peptides Or Proteins (AREA)
Abstract
提供了晚期宫颈癌的筛查方法，该方法提供用于检测HPV整合的基于分子的方法。所公开的方法允许在同一载玻片上实施Pap试验和免疫组化测试的简化方法。所公开的方法提供了廉价、高度敏感、特异的和详细的易于评估和跟进的测试。
Description
本申请要求2007年9月25日提交的美国临时申请号60/975,069、2007年12月6日提交的美国临时申请号60/992,892和2008年9月16日提交的美国申请号12/211,547作为优先权，并将这些申请的全文以引用的方式纳入本文。
领域
本公开总的涉及细胞采样、处理和筛查，以用于检测体内例如宫颈的异常组织。更具体而言，本公开涉及如下方法，从而(1)制备采集的细胞和细胞群，以检测这些细胞和细胞群的生物学特征；和(2)使用HPV-整合指示物检测人乳头瘤病毒已整合入宿主DNA的那些细胞。
背景
已广泛研究了人乳头瘤病毒(HPV)，其为发生侵入性宫颈癌和宫颈上皮内瘤样病变的前兆。宫颈癌是世界上第二常见的癌症，且是发展中国家中最常见的恶性肿瘤。
宫颈癌筛查通常基于细胞学和阴道镜分析。广泛接受的宫颈细胞学涂片(巴帕尼科拉乌试验，Pap涂片)已减少了宫颈癌的发病率和死亡率。但是，使用Pap涂片测试仍有某些缺点。常规Pap涂片的一个限制包括细胞的非典型性转移。此外，由于碎片遮蔽了异常的细胞，这些常规方法会导致假阴性和可疑结果的增加。
所有形式的Pap试验都专注于筛查高等级前兆病灶，以在这些前兆发展成侵入性癌症前开始治疗。为了有效地区分和揭示肿瘤前病灶和癌症病灶之间存在的不同的细胞学和生物学等级，报告宫颈细胞学的贝色斯达体系(Bethesda System forReporting Cervical Cytology)已提出6个类别，以对鳞状上皮细胞异常进行表征。这6个类别包括：(1)不明意义的的非典型鳞状上皮细胞(ASCUS)；(2)无法排除高等级鳞状病灶的非典型鳞状上皮细胞(ASC-H)；(3)低等级的鳞状上皮内病灶(LSIL)；(4)高等级的鳞状上皮内病灶(HSIL)；(5)鳞状上皮细胞癌(SCC)；和(6)正常。在这一体系下，HSIL可任选地被细分为“HSIL(中度发育异常，CIN II)”或“HSIL(严重发育异常，CIN III)”。已提议使用阴道镜检查和可能的活检作为ASC-H、LSIL、HSIL和SCC的后续检测，而HPV试验则提议作为ASCUS的后续检测。
HPV发病机制
存在大量的HPV类型(＞100)，但仅有一些具有嗜生殖器的性质。与那些主要在非恶性病灶中出现的HPV类型相比(这些类型被认为是低风险的类型)，已将宫颈癌中常出现的HPV类型分类为高风险类型。整合到宿主基因组中的高风险HPV DNA在HPV发病中起到关键的作用。HPV表达的基因被分成早期(E)表达基因和晚期(L)表达基因。这种整合导致E2病毒基因的特异性中断和起始于E1、E4和E5基因的其它E基因的转录和翻译。E2蛋白通常负调节E6和E7病毒致癌基因的转录，结果，E2表达的缺失导致E6和E7基因在宫颈癌细胞中的表达未被调节。
这些致癌蛋白主要通过蛋白-蛋白相互作用而起作用，使调节细胞周期进程和增殖的途径中断。E6靶向p53，使其被蛋白酶降解，结果废除了p53转录途径，导致大量正常情况下被p53抑制的基因的表达被上调。E7基因产物对成视网膜细胞瘤(Rb)基因有类似的作用。
HPV试验
HPV的自然史是，大多数的HPV感染将复发，而与治疗无关。高风险HPV类型的持续存在对于发展至HPV整合是必需的。这种发展在少部分患者中出现。因此，以高敏感性筛查HPV整合的筛查测试对于早期诊断和治疗是希望的。
HPV试验的一个挑战因素是其Pap试验结果为ASCUS的妇女。当前对于ASCUS的建议是，如果患者在30岁或30岁以上，则测试其HPV DNA存在与否。可采用几种测试来检测HPV。临床设置中最常用的测试是Digene Hc2测试，可使用标准的试剂盒来实现。该测试涉及识别特定类型的乳头瘤病毒，并提议用该测试替换常规的细胞学筛查过程，或与常规的细胞学筛查过程一起使用。
Digene Hc2测试的一个缺点是该测试通过杂交方法仅鉴别出病毒DNA的存在。由于大多数患者的HPV感染会逆转而未整合入妇女的基因组中，临床医生从阳性的Digene Hc2测试仅得知病毒是否存在。即，该测试不指示病毒是否已确实整合到妇女的基因组中。结果是，即使临床医生从Digene Hc2测试获得阳性结果，他仍无法提供确切的癌细胞确实存在的结果。此外，Digene Hc2测试复杂，花费时间和劳动，且昂贵。
到目前为止还没有针对HPV感染的治疗。因此，诊断有HPV感染的患者仅能频繁使用Pap试验监测癌症的存在。明显地，仍需要一种针对HPV整合的测试。
发明内容
提供了一种晚期宫颈癌筛查方法，该方法涉及筛查可在最早阶段治疗的病灶的细胞样品。在一个实施例中，通过一种最新型的测试过程而得以实现该方法，其中，可在Pap染色和免疫组化测试中使用一个接受结构，从而提高敏感性和特异性，并为昂贵的HPV测试方法提供低成本的替代方法。所公开的方法提供了一种基于分子的方法，该方法检测HPV是否已整合到妇女的基因组中。
在一个实施例中，用细胞采样器获取细胞，该采样器优选设计为采集细胞群或细胞团。在一个例子中，细胞群或细胞团从整个360度的宫颈腔中采集。在另一例子中，细胞采样器在采样时维持着所述群或团的空间取向，从而产生一圆形的“宫颈图”，该“宫颈图”代表细胞与宫颈解剖的空间关系，即，宫颈内膜、外子宫颈和转换带(transformation zone)的关系以及四分体的关系(quadrant relationship)。
临床医生一旦采集到细胞，可将细胞转移到接受结构中，例如载玻片。在一个实施方式中，对载玻片实施Pap染色。在一个例子中，如果结果是阳性的，可使用一种或多种生物学指示物对该相同的载玻片进行免疫染色。
所使用的生物学指示物是HPV基因蛋白的抗体。在一个例子中，这些蛋白仅在HPV已整合到患者基因组的情况下被检测到。在此例子中，免疫染色允许人们确定HPV是否已整合到患者的基因组中。在对该载玻片染色后，扫描该载玻片，以确定HPV早期(E)基因产物的抗体识别程度。
在另一实施方式中，在采集细胞并将其转移到接受结构(如载玻片)后，可将先用生物学指示物对载玻片进行免疫染色，然后扫描。然后可以决定哪份样品进行Pap染色，进行反向测试(reflux test)。
附图简述
图1A-1C阐述HPV的主要的高风险类型的E1蛋白的部分氨基酸序列。下划线所示的序列指共有序列区域，由该区域可产生在所揭示的方法的一个实施方式中用来生产诊断抗体的多肽。
图2阐述HPV的主要的高风险类型的E4蛋白的氨基酸序列。下划线所示的序列指共有序列区域，由该共有序列可产生在所揭示的方法的一个实施方式中用来生产诊断抗体的多肽。
图3阐述HPV的主要的高风险类型的E5蛋白的氨基酸序列。下划线所示的序列指共有序列区域，由该共有序列可产生在所揭示的方法的一个实施方式中用来生产诊断抗体的多肽。
图4A和4B阐述HPV的主要的高风险类型的E6蛋白的氨基酸序列。下划线所示的序列指共有序列区域，由该共有序列可产生在所揭示的方法的一个实施方式中用来生产诊断抗体的多肽。
图5阐述HPV的主要的高风险类型的E7蛋白的氨基酸序列。下划线所示的序列指共有序列区域，由该共有序列可产生在所揭示的方法的一个实施方式中用来生产诊断抗体的多肽。
图6阐述用HPV 16型的E6和E7的抗体染色的宫颈细胞。染色显示宫颈细胞对HPV整合呈阴性。
图7阐述用HPV 16型的E6和E7的抗体染色的宫颈细胞。染色显示宫颈细胞对HPV整合呈阳性。
图8阐述用HPV 16型的E6和E7的抗体染色的宫颈细胞。染色显示宫颈细胞对HPV整合呈阴性。
图9阐述用HPV 16型的E6和E7的抗体染色的宫颈细胞。染色显示宫颈细胞对HPV整合呈阳性。
详细描述
概述
提供了晚期宫颈癌筛查方法，该方法提供一基于分子的方法，用于检测HPV整合。所揭示的方法允许以简化的方法，在同一载玻片上进行Pap试验和免疫组化测试。所揭示的方法提供了廉价、高度敏感、特异的和详细的易于评估和跟进的测试。
通常，该方法包括使用采样器采集细胞群，设计该采样器，以增加其获取细胞群和促进将所采集的细胞群转移到接受结构(如载玻片)的能力。在一个例子中，使用转移装置(station)将细胞群从采样器转移到载玻片上，通过这样的方式维持采样前该细胞群内细胞之间存在的空间取向。
在一个实施方式中，在实验室中在载玻片上实施Pap染色。对所使用的Pap染色方法的类型无特别限制，可以是常规使用的Pap染色方法。
在一个例子中，如果测试是阴性的，该方法涉及按照标准的临床指导实施。如果测试是阳性的，使用与荧光或比色标签偶联的HPV整合指示物的抗体染色该载玻片。在一种情况中，所使用的HPV整合指示物是HPV基因产物(蛋白质)。对所使用的HPV基因产物的类型无特别限制，包括E1、E4、E5、E6和E7蛋白。对所使用的染色方法和指示物无特别限制，只要可实现HPV整合指示物的免疫组化定位即可。
然后扫描载玻片。对所使用的扫描仪无特别限制，可以是自动的或手控的扫描仪，或者是自动的或手控的显微镜。扫描仪用于扫描荧光或颜色信号，从而识别HPV基因产物是否存在于宫颈细胞中。
采集
使用采样器来采集细胞和细胞群。采样器可以是能采集细胞和细胞群的任何形式的采样器。合适的采样器的例子在已出版的美国专利申请US 2006/0189893和US 2006/0161076中有描述，本文将其全文以引用的方式纳入。在可使用的采样器的其它详细内容中，这两份出版物描述了采样器的材料的组合、采样器的采样表面的质地、和采样期间采样器的扩张和旋转的使用以促进细胞群的采样。采样器从宫颈的子宫颈内和子宫颈外区域获得细胞群。
接受结构
在使用接受结构时，对所使用的接受结构并无特别限制。合适的接受结构的例子包括载玻片、培养皿、膜、和其它细胞和细胞群可转移到上面以用于后续分析的结构。在一个例子中，接受结构的粘附性高于含有细胞和细胞群的采样器的表面的粘附性，以促进细胞群从采样器转移到该接受结构。当接受结构是载玻片时，可用一涂层预先处理该载玻片，以产生更大的粘附性。合适的载玻片的例子具有基于聚-L-赖氨酸的涂层，该涂层帮助细胞粘附到该载玻片上。
在另一实施方式中，不使用接受结构。在此情况中，宫颈细胞采集在固定剂中，为悬浮形式，然后直接对悬浮的细胞实施免疫学测试，而不需要使用接受结构。
Pap试验
一旦将细胞群转移到适当的接受结构(如载玻片)上，可实施常规的Pap试验。该试验涉及根据巴帕尼科拉乌方法染色载玻片和根据贝色斯达(Bethesda)系统评估。如果细胞表现正常，或者显示萎缩、组织变形或炎性变化，则细胞学视为阴性。阳性的Pap涂片定义为ASCUS或更差。
在另一实施方式中，所揭示的方法涉及在通过免疫染色实施HPV整合测试后实施一Pap试验，作为反向测试。
免疫染色
在一个例子中，一旦Pap试验证实是阳性，用一种或多种HPV整合指示物实施免疫染色。对所使用的免疫染色方法并无特别限制。例如，可实施直接或间接的免疫染色。在实施直接免疫染色的情况中，检测系统共价连接于或直接偶联于一级抗体。所使用的检测系统可经荧光标记或酶底物标记。
在使用荧光指示物的情况中，示例性的方案涉及洗涤载玻片、用封闭溶液封闭、用与荧光标记物偶联的HPV整合指示物特异性抗体染色、和然后洗涤载玻片。然后使用荧光显微镜人工读取该载玻片，或使用自动荧光扫描仪读取。扫描仪检测和定位HPV已被整合到其DNA中的细胞中的HPV整合指示物。也可通过定量评估信号来测定HPV整合指示物的表达丰度。
在使用酶-底物标记的情况中，酶必须具有底物系统，该底物系统产生可视化的产物，且产物从产生位点的扩散最小。对所使用的组化酶没有特别限制，可包括例如下述任一种酶：辣根过氧化物酶、碱性磷酸酯酶、葡萄糖氧化酶或β-半乳糖苷酶。这种类型的系统的一个示例性方案涉及用封闭溶液封闭、用与组化酶偶联的HPV整合指示物特异性抗体一起孵育、洗涤载玻片、然后用底物处理。底物可以是生色团底物，且可以是下述任一种：二氨基联苯胺(DAB)、氨乙基咔唑(AEC)、α萘酚焦宁、4-氯-1-萘酚、对苯二胺邻苯二酚、坚牢红TR、坚牢蓝BB、BCIPINBT、四唑蓝或BCl。然后用自动比色扫描仪读取载玻片，定量评估信号的亮度。或者，使用标准的显微镜观察载玻片，并鉴定该分子指示物的存在以及细胞的任何形态学变化。
在使用间接染色的情况中，示例性的方案涉及用对HPV整合指示物特异的未标记的一级抗体培育、洗涤、用与组化酶标记偶联的二级抗体培育、接着洗涤和检测。
免疫组化染色的定量评估
对定量评估指示物的表达丰度的方法无特别限制。在免疫荧光染色的情况中，可用图像分析软件(如Analyze)来评估扫描得到的组织学图像。该软件允许人们确定信号密度，并计算亮度-面积-产物(BAP)。BAP可提供一手段，用于客观地比较暴露于相同染色方案的同一解剖区域中染色的强度。可采用以下公式计算BAP：
(范围内平均像素亮度-范围内最小亮度)×(范围内像素数量)
在使用比色标签的情况中，使用单片CCD红-绿-蓝(RGB)颜色摄像机和标准的诊断显微镜将扫描的图像输入计算机的S-VHS端口。用于评价颜色信号的软件可以是例如Photoshop，但不限于此。如果使用Photoshop，使用例如Histogram命令定量信号，以测量色调分布，作为自动图像操作的基础。此特征允许获得所使用的特定底物(例如，生色团)的空间信息，且此特征可表示为整个图像的百分比，或者以μm2表示。
在另一实施方式中，可通过计算阳性细胞的比例而定量比色标签。
HPV整合指示物
在一个实施方式中，在所揭示的方法中使用的抗体对HPV整合指示物特异。术语“HPV整合指示物特异性抗体”在本文中指抗HPV基因的蛋白产物的表位的特异性抗体，所述表位例如产生自HPV整合的基因产物的表位，包括但不限于E1、E4、E5、E6和/或E7。术语“表位”在本文中指B和/或T细胞对其响应的抗原上的位点。抗原可以是多肽或蛋白产物。位点可以是多肽。在一个例子中，表位可以是抗原，也可以指HPV整合指示物的多肽。
虽然抗体大多数情况下仅对一种蛋白或多肽特异，但本发明者的目的是使用识别来自任意高风险的HPV类型的蛋白质家族的抗体。为此，本发明者已选择高风险HPV类型各自的感兴趣的蛋白之间的共有区域。图1A-1C显示E1蛋白的高度保守的区域。图2显示E4蛋白的高度保守区域。图3显示E5蛋白的高度保守区域。图4A和4B显示E6蛋白的高度保守区域。图5显示E7蛋白的高度保守区域。
抗体的制备
可使用本领域充分记载的各种标准免疫技术，通过注射然后强化注射抗原混合物而在任何合适的动物(例如，小鼠和兔)中产生抗体。为了制备单克隆抗体组，杀死免疫的动物，收集脾细胞，使其无限繁殖，以获得能够产生所需抗体的细胞。用所述注射的多肽筛查这些细胞的上清，作为感兴趣抗体存在的第一次测试。
实施例1
在一个实施例中，使用HPV E1基因产生针对多肽的抗体。用于产生抗体的多肽的具体例子如下：
在HPV 16型的位置202：SNAKAMLAKFKELYGC(SEQ ID NO：1)和在HPV18型的位置208：NNKQGAMLAVFKDTYGC(SEQ ID NO：2)；在HPV 16型的位置293：LSKLLCVSPMC(ACM)MMIEPPKLR(SEQ ID NO：3)和在HPV 18型的位置301：LSTLLHVPETAMLIEPPKLR(SEQ ID NO：4)和在HPV 33、52和58型的位置286：LSSLLNIPQSQMLIQPPKLR(SEQ ID NO：5)；和在HPV 16型的位置606：DKNWKSFFSRTWC(SEQ ID NO：6)和在HPV 18型的位置613：DKNWKCFFERTWC(SEQ ID NO：7)和在HPV 26、51、66和73型的位置587：NENWKAFFTKTWC(SEQ ID NO：8)；和在HPV 16型的位置640：CVSGQNTNTL(SEQ ID NO：9)和在HPV 18型的位置648：CLRAGQNHRPL(SEQ ID NO：10)和在HPV 26、31、33、39、45、51、52、58和59型的位置620：CSTGENIRSI(SEQ IDNO：11)。
在另一例子中，使用HPV E4基因产生针对多肽的抗体。用于产生抗体的多肽的具体例子如下：
HPV 16型的位置44：RRLSSDQD(SEQ ID NO：12)，HPV 16型的位置79：LTAHQTK(SEQ ID NO：13)，和HPV 16型的位置85：DGLTVIVTL(SEQ ID NO：14)。
在另一例子中，使用HPV E5基因产生针对多肽的抗体。用于产生抗体的多肽的具体例子如下：
HPV 52、58和33型的位置5：VFC(ACM)FILFLC(ACM)LC(SEQ ID NO：15)和HPV 35型的位置15：LC(ACM)FC(ACM)VLLC(ACM)LCL(SEQ ID NO：16)，和HPV 18型的位置30：AYAWVLVFVYIVV(SEQ ID NO：17)，和
HPV 51和69型的位置31：PLLLSQYVFAAHLLLII(SEQ ID NO：18)，和
HPV 16型的位置37：STYTSLIOLV(SEQ ID NO：19)，和HPV 73型的位置51：FFLYVLVFYIF(SEQ ID NO：20)。
在另一例子中，使用HPV E6基因产生针对多肽的抗体。用于产生抗体的多肽的具体例子如下：
HPV 16型的位置126：LDKKRRFHNI(SEQ ID NO：21)和LNEKKRFHNI(SEQ ID NO：22)和LDKKQRFHNI(SEQ ID NO：23)，HPV 16型的位置129：KQRFHNIRGRWTGRC(SEQ ID NO：24)和KRRFHNIAGRYTGQC(SEQ ID NO：25)和NKRFHNIRGRWTGRC(SEQ ID NO：26)。
在另一实施例中，使用HPV E7基因产生针对多肽的抗体。用于产生抗体的多肽的具体例子如下：
HPV 16型的位置29：DSSEENDEID(SEQ ID NO：27)，和HPV 16型的位置41：PAGGA(SEQ ID NO：28)，和HPV 16型的位置89：VCPIC(SEQ ID NO：29)
设计上述多肽，以使它们的抗体能够组合使用来识别单一检测混合物中所有的高风险HPV类型。
实施例2
所揭示的方法的一个例子包括固定宫颈细胞，室温以1000RPM细胞离心(cyto-centriguge)2分钟而在载玻片上沉积细胞，在0.01M柠檬酸盐缓冲液(pH6.0)中培育载玻片30分钟，在TBST(Tris缓冲的盐水，含Tween 20)中洗涤载玻片5分钟，用正常胎牛血清封闭缓冲液(10％FCS的TBST溶液)培育载玻片10分钟，用一级单克隆小鼠抗体的稀释缓冲液(1FCS的TBST溶液)以1∶50、1∶75或1∶100的稀释度培育1小时(最佳稀释度根据各抗体而变)，在TBST中洗涤载玻片5分钟，用与HRP(辣根过氧化物酶)偶联的二级兔抗小鼠抗体培育，和然后用HRP酶的底物DAB(二氨基联苯胺生色团)培育10分钟，在蒸馏水中清洗载玻片1分钟，干燥后滴入1滴DAPI核染剂。这在含有一级抗体与之反应的抗原的细胞中产生了褐色。然后可使用亮视野显微镜对250个细胞进行计数，确定被阳性染色的细胞的百分比。
对一系列的从实施了阴道镜检测、因而更有可能具有HPV整合和宫颈病灶的患者得到的临床样品使用这种方法，本发明者已发现了下表以及图6-9所示的结果。
| 样品标示 | HPV 16 E6抗体阳性％ | HPV 16 E7抗体阳性％ | 两种抗体都呈阳性的％ |
| ARR 2 | 12 | 18 | 17 |
| ARR 14 | 0 | 2 | 2 |
| ARR 16 | 0 | 0 | 0 |
| ARR 18 | 7 | 4 | 4 |
| ARR 19 | 0 | 0 | 0 |
| ARR 22 | 0 | 1 | 0 |
| ARR 24 | 0 | 3 | 0 |
| GA 1 | 0 | 0 | 0 |
| GA 3 | 34 | 51 | 38 |
| GA 9 | 0 | 0 | 0 |
| GA 12 | 0 | 0 | 0 |
| GA 19 | 8 | 6 | 5 |
| GA 20 | 0 | 1 | 2 |
| GA 21 | 0 | 2 | 1 |
| GA 24 | 0 | 3 | 0 |
| GA 25 | 2 | 3 | 2 |
| GA 26 | 0 | 0 | 0 |
| GA 31 | 0 | 1 | 1 |
| GA 33 | 19 | 25 | 21 |
| WHR 2 | 0 | 1 | 1 |
以上以及图6-9所示的结果表明，对于单个患者，可容易地区分哪些患者具有整合的HPV而哪些没有。这些结果也表明，对于E6和E7抗体，单独使用它们获得的信息与组合使用它们所获得的信息相同。最后，值得注意的是，5/20(25％)的这些患者对HPV 16型整合呈阳性，但他们来自于已知具有异常Pap试验的患者亚组。
实施方式
任意大小或组织形式的实验室都可实施所述的晚期宫颈癌筛查方法。较佳地，由已实施了Pap试验的实验室或护理者实施。
所述晚期宫颈癌筛查方法传递一种独特的采集方法，该采集方法为患者的改进管理提供了宫颈癌筛查的增值服务。这些方法涉及以简化的方法通过Pap试验和免疫组化染色对细胞学样品进行分析，从而不仅对真阳性和假阳性进行有效地鉴定，而且也对真阴性和假阴性进行有效地鉴定。与常规的方法相比，本方法通过单细胞采集方法为医师或外科医生提供更明确和详细的结果。优点包括：改进的阳性和阴性预测值，因为指示物指示何时已发生了病毒整合入妇女基因组；专有的采集方法，该方法使患者在细胞采集过程中感到更舒适；低成本方案；具有高的均匀性、质量和完整性的过剩细胞的采集，因为细胞体积允许对单次样品采集获得的细胞实施多次筛查试验；和简化的试验方法，其中单个载玻片可用于Pap染色和免疫细胞化学试验两者。
本发明方法为实验室提供了益处，因为这些方法在实施昂贵的特异性HR-HPV测试方法前提供低成本的替代的方式；通过提供敏感性和特异性提高的测试方法而帮助消除假阳性，并对ASCUS样品正确分类；为进入宫颈癌分子诊断市场提供了切入口，对于当前和新的消费者而言扩大了复杂诊断服务的选择(portfolio)。
还清楚的是，可将上述方法作为宫颈癌的基本筛查试验，而不需要实施Pap试验。
虽然已结合示例性实施方式描述了本发明，但对于本领域技术人员来说显而易见的是，在权利要求书所限定的本发明范围之内还可对本发明作出其他的变动。所公开的细节并不是对本发明的限制。
序列表
<110>坎维耶股份有限公司(CANVIR，INC.)
<120>晚期宫颈细胞筛查方法
<130>20041.21WOU1
<150>US 12/211,547
<151>2008-09-16
<150>US 60/975069
<151>2007-09-25
<150>US 60/992892
<151>2007-12-06
<160>122
<170>PatentIn version 3.5
<210>1
<211>16
<212>PRT
<213>人乳头瘤病毒16型
<400>1
Ser Asn Ala Lys Ala Met Leu Ala Lys Phe Lys Glu Leu Tyr Gly Cys
1 5 10 15
<210>2
<211>17
<212>PRT
<213>人乳头瘤病毒16型
<400>2
Asn Asn Lys Gln Gly Ala Met Leu Ala Val Phe Lys Asp Thr Tyr Gly
1 5 10 15
Cys
<210>3
<211>23
<212>PRT
<213>人乳头瘤病毒16型
<400>3
Leu Ser Lys Leu Leu Cys Val Ser Pro Met Cys Ala Cys Met Met Met
1 5 10 15
Ile Glu Pro Pro Lys Leu Arg
20
<210>4
<211>20
<212>PRT
<213>人乳头瘤病毒16型
<400>4
Leu Ser Thr Leu Leu His Val Pro Glu Thr Ala Met Leu Ile Glu Pro
1 5 10 15
Pro Lys Leu Arg
20
<210>5
<211>20
<212>PRT
<213>人乳头瘤病毒16型
<400>5
Leu Ser Ser Leu Leu Asn Ile Pro Gln Ser Gln Met Leu Ile Gln Pro
1 5 10 15
Pro Lys Leu Arg
20
<210>6
<211>13
<212>PRT
<213>人乳头瘤病毒16型
<400>6
Asp Lys Asn Trp Lys Ser Phe Phe Ser Arg Thr Trp Cys
1 5 10
<210>7
<211>13
<212>PRT
<213>人乳头瘤病毒16型
<400>7
Asp Lys Asn Trp Lys Cys Phe Phe Glu Arg Thr Trp Cys
1 5 10
<210>8
<211>13
<212>PRT
<213>人乳头瘤病毒16型
<400>8
Asn Glu Asn Trp Lys Ala Phe Phe Thr Lys Thr Trp Cys
1 5 10
<210>9
<211>10
<212>PRT
<213>人乳头瘤病毒16型
<400>9
Cys Val Ser Gly Gln Asn Thr Asn Thr Leu
1 5 10
<210>10
<211>11
<212>PRT
<213>人乳头瘤病毒16型
<400>10
Cys Leu Arg Ala Gly Gln Asn His Arg Pro Leu
1 5 10
<210>11
<211>10
<212>PRT
<213>人乳头瘤病毒16型
<400>11
Cys Ser Thr Gly Glu Asn Ile Arg Ser Ile
1 5 10
<210>12
<211>8
<212>PRT
<213>人乳头瘤病毒16型
<400>12
Arg Arg Leu Ser Ser Asp Gln Asp
1 5
<210>13
<211>7
<212>PRT
<213>人乳头瘤病毒16型
<400>13
Leu Thr Ala His Gln Thr Lys
1 5
<210>14
<211>9
<212>PRT
<213>人乳头瘤病毒16型
<400>14
Asp Gly Leu Thr Val Ile Val Thr Leu
1 5
<210>15
<211>11
<212>PRT
<213>人乳头瘤病毒52型
<400>15
Val Phe Cys Phe Ile Leu Leu Met Val Phe Cys
1 5 10
<210>16
<211>11
<212>PRT
<213>人乳头瘤病毒35型
<400>16
Leu Cys Leu Cys Val Cys Leu Val Leu Cys Cys
1 5 10
<210>17
<211>13
<212>PRT
<213>人乳头瘤病毒18型
<400>17
Ala Tyr Ala Trp Val Leu Val Phe Val Tyr Ile Val Val
1 5 10
<210>18
<211>16
<212>PRT
<213>人乳头瘤病毒51型
<400>18
Pro Leu Leu Leu Ser Gln Tyr Val Phe Ala Ala Ala Leu Leu Leu Ile
1 5 10 15
<210>19
<211>10
<212>PRT
<213>人乳头瘤病毒16型
<400>19
Ser Thr Tyr Thr Ser Leu Ile Ile Leu Val
1 5 10
<210>20
<211>11
<212>PRT
<213>人乳头瘤病毒73型
<400>20
Phe Phe Leu Tyr Val Leu Val Phe Tyr Ile Pro
1 5 10
<210>21
<211>10
<212>PRT
<213>人乳头瘤病毒16型
<400>21
Leu Asp Lys Lys Gln Arg Phe His Asn Ile
1 5 10
<210>22
<211>10
<212>PRT
<213>人乳头瘤病毒18型
<400>22
Leu Asn Glu Lys Arg Arg Phe His Asn Ile
1 5 10
<210>23
<211>10
<212>PRT
<213>人乳头瘤病毒16型
<400>23
Leu Asp Lys Lys Arg Arg Phe His Asn Ile
1 5 10
<210>24
<211>13
<212>PRT
<213>人乳头瘤病毒16型
<400>24
Arg Phe His Asn Ile Arg Gly Arg Trp Thr Gly Arg Cys
1 5 10
<210>25
<211>15
<212>PRT
<213>人乳头瘤病毒18型
<400>25
Lys Arg Arg Phe His Asn Ile Ala Gly Arg Tyr Thr Gly Gln Cys
1 5 10 15
<210>26
<211>15
<212>PRT
<213>人乳头瘤病毒52型
<400>26
Asn Lys Arg Phe His Asn Ile Arg Gly Arg Trp Thr Gly Arg Cys
1 5 10 15
<210>27
<211>10
<212>PRT
<213>人乳头瘤病毒16型
<400>27
Asp Ser Ser Glu Glu Asn Asp Glu Ile Asp
1 5 10
<210>28
<211>5
<212>PRT
<213>人乳头瘤病毒16型
<400>28
Pro Ala Gly Gly Ala
1 5
<210>29
<211>5
<212>PRT
<213>人乳头瘤病毒16型
<400>29
Val Cys Pro Ile Cys
1 5
<210>30
<211>30
<212>PRT
<213>人乳头瘤病毒16型
<400>30
Leu Lys Thr Ser Asn Ala Lys Ala Ala Met Leu Ala Lys Phe Lys Glu
1 5 10 15
Leu Tyr Gly Val Ser Phe Ser Glu Leu Val Arg Pro Phe Lys
20 25 30
<210>31
<211>30
<212>PRT
<213>人乳头瘤病毒16型
<400>31
Glu Thr Ile Glu Lys Leu Leu Ser Lys Leu Leu Cys Val Ser Pro Met
1 5 10 15
Cys Met Met Ile Glu Pro Pro Lys Leu Arg Ser Thr Ala Ala
20 25 30
<210>32
<211>30
<212>PRT
<213>人乳头瘤病毒18型
<400>32
Leu Lys Val Asn Asn Lys Gln Gly Ala Met Leu Ala Val Phe Lys Asp
1 5 10 15
Thr Tyr Gly Leu Ser Phe Thr Asp Leu Val Arg Asn Phe Lys
20 25 30
<210>33
<211>30
<212>PRT
<213>人乳头瘤病毒18型
<400>33
Leu Thr Val Ala Lys Gly Leu Ser Thr Leu Leu His Val Pro Glu Thr
1 5 10 15
Cys Met Leu Ile Gln Pro Pro Lys Leu Arg Ser Ser Val Ala
20 25 30
<210>34
<211>30
<212>PRT
<213>人乳头瘤病毒26型
<400>34
Leu Lys Cys Ser Asn Val Lys Ala Ala Leu Leu Ser Lys Phe Lys Thr
1 5 10 15
Val Tyr Gly Val Ser Phe Ala Glu Leu Val Arg Val Phe Lys
20 25 30
<210>35
<211>30
<212>PRT
<213>人乳头瘤病毒26型
<400>35
Thr Thr Ile Lys Asn Cys Leu Cys Met Leu Leu Asn Val Pro Glu Thr
1 5 10 15
Gln Leu Leu Ile Glu Pro Pro Lys Leu Arg Ser Thr Ala Val
20 25 30
<210>36
<211>30
<212>PRT
<213>人乳头瘤病毒31型
<400>36
Leu Lys Thr Ser Asn Gly Lys Ala Ala Met Leu Gly Lys Phe Lys Glu
1 5 10 15
Leu Tyr Gly Val Ser Phe Met Glu Leu Ile Arg Pro Phe Gln
20 25 30
<210>37
<211>30
<212>PRT
<213>人乳头瘤病毒31型
<400>37
Ile Thr Ile Glu Lys Leu Leu Glu Lys Leu Leu Cys Ile Ser Thr Asn
1 5 10 15
Cys Met Leu Ile Gln Pro Pro Lys Leu Arg Ser Thr Ala Ala
20 25 30
<210>38
<211>30
<212>PRT
<213>人乳头瘤病毒33型
<400>38
Leu His Ser Ser Asn Thr Lys Ala Asn Ile Leu Tyr Lys Phe Lys Glu
1 5 10 15
Ala Tyr Gly Ile Ser Phe Met Glu Leu Val Arg Pro Phe Lys
20 25 30
<210>39
<211>30
<212>PRT
<213>人乳头瘤病毒33型
<400>39
Leu Thr Val Ala Lys Leu Met Ser Asn Leu Leu Ser Ile Pro Glu Thr
1 5 10 15
Cys Met Val Ile Glu Pro Pro Lys Leu Arg Ser Gln Thr Cys
20 25 30
<210>40
<211>30
<212>PRT
<213>人乳头瘤病毒35型
<400>40
Leu Lys Cys Ser Asn Ala Asn Ala Ala Met Leu Ala Lys Phe Lys Glu
1 5 10 15
Leu Phe Gly Ile Ser Phe Thr Glu Leu Ile Arg Pro Phe Lys
20 25 30
<210>41
<211>30
<212>PRT
<213>人乳头瘤病毒35型
<400>41
Thr Thr Ile Glu Lys Leu Leu Ser Lys Leu Leu Cys Ile Ser Ala Ala
1 5 10 15
Ser Met Leu Ile Gln Pro Pro Lys Leu Arg Ser Thr Pro Ala
20 25 30
<210>42
<211>30
<212>PRT
<213>人乳头瘤病毒39型
<400>42
Leu Gln Ser Asn Asn Lys Lys Ala Ala Met Leu Thr Gln Phe Lys Glu
1 5 10 15
Thr Tyr Gly Leu Ser Phe Thr Asp Leu Val Arg Thr Phe Lys
20 25 30
<210>43
<211>30
<212>PRT
<213>人乳头瘤病毒39型
<400>43
Val Thr Val Gly Lys Gly Leu Ser Thr Leu Leu His Val Pro Glu Ser
1 5 10 15
Cys Met Leu Leu Glu Pro Pro Lys Leu Arg Ser Pro Val Ala
20 25 30
<210>44
<211>30
<212>PRT
<213>人乳头瘤病毒45型
<400>44
Leu Gln Ala Ser Asn Lys Lys Ala Ala Met Leu Ala Val Phe Lys Asp
1 5 10 15
Ile Tyr Gly Leu Ser Phe Thr Asp Leu Val Arg Asn Phe Lys
20 25 30
<210>45
<211>30
<212>PRT
<213>人乳头瘤病毒45型
<400>45
Leu Thr Val Ala Lys Gly Leu Ser Thr Leu Leu His Val Pro Glu Thr
1 5 10 15
Cys Met Leu Ile Glu Pro Pro Lys Leu Arg Ser Ser Val Ala
20 25 30
<210>46
<211>30
<212>PRT
<213>人乳头瘤病毒51型
<400>46
Leu Lys Ser Ser Asn Ala Lys Ala Thr Leu Met Ala Lys Phe Lys Glu
1 5 10 15
Leu Tyr Gly Ile Ser Tyr Asn Glu Leu Val Arg Val Phe Lys
20 25 30
<210>47
<211>30
<212>PRT
<213>人乳头瘤病毒51型
<400>47
Thr Thr Ile Ala Lys Cys Leu Ser Thr Leu Val Asn Ile Pro Gln Ser
1 5 10 15
Gln Met Phe Ile Glu Pro Pro Lys Leu Arg Ser Thr Pro Val
20 25 30
<210>48
<211>30
<212>PRT
<213>人乳头瘤病毒52型
<400>48
Met Cys Glu Asn Ser Ile Lys Thr Thr Val Leu Phe Lys Phe Lys Glu
1 5 10 15
Thr Tyr Gly Val Ser Phe Met Glu Leu Val Arg Pro Phe Lys
20 25 30
<210>49
<211>30
<212>PRT
<213>人乳头瘤病毒52型
<400>49
Leu Thr Val Ser Lys Leu Met Ser Gln Leu Leu Asn Ile Pro Glu Thr
1 5 10 15
His Met Val Ile Glu Pro Pro Lys Leu Arg Ser Ala Thr Cys
20 25 30
<210>50
<211>30
<212>PRT
<213>人乳头瘤病毒58型
<400>50
Leu His Asn Ser Asn Thr Lys Ala Thr Leu Leu TyrLys Phe Lys Glu
1 5 10 15
Ala Tyr Gly Val Ser Phe Met Glu Leu Val Arg Pro Phe Lys
20 25 30
<210>51
<211>30
<212>PRT
<213>人乳头瘤病毒58型
<400>51
Leu Thr Val Ala Lys Leu Met Ser Asn Leu Leu Ser Ile Pro Glu Thr
1 5 10 15
Cys Met Ile Ile Glu Pro Pro Lys Leu Arg Ser Gln Ala Cys
20 25 30
<210>52
<211>30
<212>PRT
<213>人乳头瘤病毒59型
<400>52
Leu His Ser Lys Asn Lys Lys Ala Ala Met Tyr Ala Lys Phe Lys Glu
1 5 10 15
Leu Tyr Gly Leu Ser Phe Gln Asp Leu Val Arg Thr Phe Lys
20 25 30
<210>53
<211>30
<212>PRT
<213>人乳头瘤病毒59型
<400>53
Ile Thr Val Ala Lys Gly Leu Ser Thr Leu Leu His Val Pro Asp Thr
1 5 10 15
Cys Met Leu Ile Glu Pro Pro Lys Leu Arg Ser Gly Val Ala
20 25 30
<210>54
<211>30
<212>PRT
<213>人乳头瘤病毒66型
<400>54
Phe Lys Ser Ser Asn Val Gln Gly Arg Leu His Phe Lys Phe Lys Glu
1 5 10 15
Val Tyr Gly Val Pro Tyr Thr Glu Leu Val Arg Thr Phe Lys
20 25 30
<210>55
<211>30
<212>PRT
<213>人乳头瘤病毒66型
<400>55
Lys Thr Ile Thr Lys Ser Leu Ser Ser Ile Leu Asn Val Pro Gln Glu
1 5 10 15
Gln Met Leu Ile Gln Pro Pro Lys Leu Arg Ser Pro Ala Val
20 25 30
<210>56
<211>30
<212>PRT
<213>人乳头瘤病毒73型
<400>56
Leu Gln Arg Asn Asn Ala Lys Ala Ala Leu Leu Ala Lys Phe Lys Glu
1 5 10 15
Val Tyr Gly Leu Ser Tyr Met Glu Leu Val Arg Pro Tyr Lys
20 25 30
<210>57
<211>30
<212>PRT
<213>人乳头瘤病毒73型
<400>57
Leu Thr Val Gln Lys Leu Leu Ser Ser Leu Leu Asn Val Thr Gln Glu
1 5 10 15
Arg Met Leu Ile Glu Pro Pro Arg Leu Arg Ser Thr Pro Cys
20 25 30
<210>58
<211>52
<212>PRT
<213>人乳头瘤病毒16型
<400>58
Gly Asn Pro Val Tyr Glu Leu Asn Asp Lys Asn Trp Lys Ser Phe Phe
1 5 10 15
Ser Arg Thr Trp Ser Arg Leu Ser Leu His Glu Asp Glu Asp Lys Glu
20 25 30
Asn Asp Gly Asp Ser Leu Pro Thr Phe Lys Cys Val Ser Gly Gln Asn
35 40 45
Thr Asn Thr Leu
50
<210>59
<211>53
<212>PRT
<213>人乳头瘤病毒18型
<400>59
Gly Asn Pro Val Tyr Glu Ile Asn Asp Lys Asn Trp Lys Cys Phe Phe
1 5 10 15
Glu Arg Thr Trp Ser Arg Leu Asp Leu His Glu Glu Glu Glu Asp Ala
20 25 30
Asp Thr Glu Gly Asn Pro Phe Gly Thr Phe Lys Leu Arg Ala Gly Gln
35 40 45
Asn His Arg Pro Leu
50
<210>60
<211>51
<212>PRT
<213>人乳头瘤病毒26型
<400>60
Gly Asn Pro Val Tyr Ala Leu Thr Asp Val Asn Trp Lys Ser Phe Phe
1 5 10 15
Ser Thr Thr Trp Ser Arg Leu Asp Leu Glu Glu Asp Ala Asp Lys Glu
20 25 30
Asn Gly Glu Pro Leu Pro Ala Phe Lys Cys Val Pro Gly Glu Asn Thr
35 40 45
Arg Leu Leu
50
<210>61
<211>52
<212>PRT
<213>人乳头瘤病毒31型
<400>61
Gly Asn Pro Val Tyr Glu Leu Ser Asp Lys Asn Trp Lys Ser Phe Phe
1 5 10 15
Ser Arg Thr Trp Cys Arg Leu Asn Leu His Glu Glu Glu Asp Lys Glu
20 25 30
Asn Asp Gly Asp Ser Phe Ser Thr Phe Lys Cys Val Ser Gly Gln Asn
35 40 45
Ile Arg Thr Leu
50
<210>62
<211>54
<212>PRT
<213>人乳头瘤病毒33型
<400>62
Gly Asn Pro Val Tyr Ala Ile Asn Asp Glu Asn Trp Lys Ser Phe Phe
1 5 10 15
Ser Arg Thr Trp Cys Lys Leu Asp Leu Ile Glu Glu Glu Asp Lys Glu
20 25 30
Asn His Gly Gly Asn Ile Ser Thr Phe Lys Cys Ser Ala Gly Glu Asn
35 40 45
Thr Arg Ser Leu Arg Ser
50
<210>63
<211>54
<212>PRT
<213>人乳头瘤病毒35型
<400>63
Gly Asn Pro Val Tyr Gly Leu Asn Asp Lys Asn Trp Lys Ser Phe Phe
1 5 10 15
Ser Arg Thr Trp Cys Arg Leu Asn Leu His Glu Glu Glu Asp Lys Glu
20 25 30
Asn Asp Gly Asp Ala Phe Pro Ala Phe Lys Cys Val Ser Gly Gln Asn
35 40 45
Thr Arg Thr Leu Arg Asp
50
<210>64
<211>53
<212>PRT
<213>人乳头瘤病毒39型
<400>64
Arg Asn Pro Val Tyr Thr Ile Asn Asp Lys Asn Trp Lys Cys Phe Phe
1 5 10 15
Glu Lys Thr Trp Cys Arg Leu Asp Leu Gln Gln Asp Glu Asp Glu Gly
20 25 30
Asp Asn Asp Glu Asn Thr Phe Thr Thr Phe Lys Cys Val Thr Gly Gln
35 40 45
Asn Thr Arg Ile Leu
50
<210>65
<211>53
<212>PRT
<213>人乳头瘤病毒45型
<400>65
GlyAsn Pro Val Tyr Glu Ile Asn Asp Lys Asn Trp Lys Cys Phe Phe
1 5 10 15
Glu Arg Thr Trp Ser Arg Leu Asp Leu His Glu Asp Asp Glu Asp Ala
20 25 30
Asp Thr Glu Gly Ile Pro Phe Gly Thr Phe Lys Cys Val Thr Gly Gln
35 40 45
Asn Thr Arg Pro Leu
50
<210>66
<211>51
<212>PRT
<213>人乳头瘤病毒51型
<400>66
Gly Asn Ala Val Tyr Thr Leu Asn Asp Glu Asn Trp Lys Asn Phe Phe
1 5 10 15
Ser Thr Thr Trp Ser Arg Leu Asp Leu Glu Glu Glu Glu Asp Lys Glu
20 25 30
Asn Gly Asp Pro Met Pro Pro Phe Lys Cys Val Pro Gly Glu Asn Thr
35 40 45
Arg Leu Leu
50
<210>67
<211>54
<212>PRT
<213>人乳头瘤病毒52型
<400>67
Gly Asn Pro Ile Tyr Glu Ile Asn Asn Glu Asn Trp Lys Ser Phe Phe
1 5 10 15
Ser Arg Thr Trp Cys Lys Leu Asp Leu Ile Gln Glu Glu Asp Lys Glu
20 25 30
Asn Asp Gly Val Asp Thr Gly Thr Phe Lys Cys Ser Ala Gly Lys Asn
35 40 45
Thr Arg Ser Ile Arg Ser
50
<210>68
<211>54
<212>PRT
<213>人乳头瘤病毒58型
<400>68
Gly Asn Pro Val Tyr Lys Ile Asn Asp Glu Asn Trp Lys Ser Phe Phe
1 5 10 15
Ser Arg Thr Trp Cys Lys Leu Gly Leu Ile Glu Glu Glu Asp Lys Glu
20 25 30
Asn Asp Gly Gly Asn Ile Ser Thr Phe Lys Cys Ser Ala Gly Gln Asn
35 40 45
Pro Arg His Ile Arg Ser
50
<210>69
<211>53
<212>PRT
<213>人乳头瘤病毒59型
<400>69
Arg Asn Pro Val Tyr Thr Ile Asn Asp Arg Asn Trp Lys Cys Phe Phe
1 5 10 15
Glu Arg Thr Trp Cys Arg Leu Asp Leu Asn Glu Glu Glu Glu Asp Ala
20 25 30
Asp Ser Asp Gly His Pro Phe Ala Ala Phe Lys Cys Val Thr Gly Ser
35 40 45
Asn Ile Arg Thr Leu
50
<210>70
<211>52
<212>PRT
<213>人乳头瘤病毒66型
<400>70
Gly Asn Pro Val Tyr Glu Leu Ser Asn Val Asn Trp Lys Cys Phe Phe
1 5 10 15
Glu Arg Thr Trp Ser Arg Leu Asn Leu Asp Asn Asp Glu Asp Lys Glu
20 25 30
Asn Asn Gly Asp Ser Ile Pro Thr Phe Arg Cys Val Pro Glu Gln Asn
35 40 45
Thr Arg Leu Leu
50
<210>71
<211>52
<212>PRT
<213>人乳头瘤病毒73型
<400>71
Gly Asn Pro Leu Tyr Gln Leu Thr Asn Glu Asn Trp Lys Ala Phe Phe
1 5 10 15
Thr Lys Thr Trp Ser Lys Leu Asp Leu Thr Glu Asp Asp Asp Lys Glu
20 25 30
Asn Asp Gly Asp Thr Val Gln Thr Phe Lys Cys Val Ser Gly Arg Asn
35 40 45
Pro Arg Thr Val
50
<210>72
<211>91
<212>PRT
<213>人乳头瘤病毒54/52型
<400>72
Leu His Leu Tyr Leu Val Pro Lys Arg His Cys Gln Tyr Pro Leu Leu
1 5 10 15
Ala Leu Leu Asn Thr Pro Asp Gln Pro Ile Pro His His Val Pro Thr
20 25 30
Thr Pro Gln Lys Gln Ser Arg Ala Arg Arg Arg Leu Glu Asn Glu Leu
35 40 45
Glu Ser Thr Ala Gln Thr Ser Asn His Thr Ala Pro Gln Thr Pro Trp
50 55 60
Ala Val Thr Thr Thr Gly Thr Ser Val Thr Ile Thr Thr Arg Thr Lys
65 70 75 80
Asp Gly Thr Gln Val Val Val Thr Leu His Leu
85 90
<210>73
<211>85
<212>PRT
<213>人乳头瘤病毒58/33型
<400>73
Leu His Leu Tyr Leu Val Ile Lys Tyr Pro Leu Leu Lys Leu Leu Thr
1 5 10 15
Gln Arg Pro Pro Arg Pro Pro Thr Thr Lys Val His Arg Gly Gln Ser
20 25 30
Asp Asp Asp Ser Ile Tyr Gln Thr Pro Glu Thr Thr Pro Ser Thr Pro
35 40 45
Gln Ser Ile Gln Thr Ala Pro Trp Thr Val Asp His Glu Glu Glu Asp
50 55 60
Tyr Thr Val Gln Leu Thr Val His Thr Lys Gly Gly Thr Cys Val Val
65 70 75 80
Leu Lys Phe His Leu
85
<210>74
<211>92
<212>PRT
<213>人乳头瘤病毒16型
<400>74
Leu His Leu Cys Leu Ala Ala Thr Lys Tyr Pro Leu Leu Lys Leu Leu
1 5 10 15
Gly Ser Thr Trp Pro Thr Thr Pro Pro Arg Pro Ile Pro Lys Pro Ser
20 25 30
Pro Trp Ala Pro Lys Lys His Arg Arg Leu Ser Ser Asp Gln Asp Gln
35 40 45
Ser Gln Thr Pro Glu Thr Pro Ala Thr Pro Leu Ser Cys Cys Thr Glu
50 55 60
Thr Gln Trp Thr Val Leu Gln Ser Ser Leu His Leu Thr Ala His Thr
65 70 75 80
Lys Asp Gly Leu Thr Val Ile Val Thr Leu His Pro
85 90
<210>75
<211>99
<212>PRT
<213>人乳头瘤病毒31型
<400>75
Leu Asn Leu Tyr Leu Ala Val Thr Lys Tyr Pro Leu Leu Gly Leu Leu
1 5 10 15
Gln Ser Tyr Gln Gln Pro Thr Thr Pro Pro His Arg Ile Pro Lys Pro
20 25 30
Ala Pro Trp Ala Pro Val Lys Val Cys Gly Gly Arg Arg Arg Leu Leu
35 40 45
Ser Asp Gln Glu Gln Ser Gln Ser Thr Glu Thr Pro Thr Thr Pro Thr
50 55 60
Ser Cys Cys Glu Ala Thr Pro Trp Thr Val Ser Thr Val Gly Leu Ser
65 70 75 80
Val Gln Leu His Ala Gln Thr Lys Gln Gly Leu Ser Val Val Leu Gln
85 90 95
Leu His Leu
<210>76
<211>93
<212>PRT
<213>人乳头瘤病毒35型
<400>76
Leu Asn Leu Tyr Leu Ala Ala Gln Asn Tyr Pro Leu Leu Lys Leu Leu
1 5 10 15
His Ser Tyr Thr Pro Thr Thr Pro Pro Arg Pro Ile Pro Lys Pro Ala
20 25 30
Pro Trp Ala Pro Gln Lys Pro Arg Arg Gln Ile Thr Asn Asp Phe Glu
35 40 45
Gly Val Pro Ser Ser Pro Thr Thr Pro Pro Ser Glu Cys Asp Ser Val
50 55 60
Pro Trp Thr Val Leu Thr Glu Gly Ser Thr Leu His Leu Thr Ala Gln
65 70 75 80
Thr Lys Thr Gly Val Val Val Val Val Gln Leu His Leu
85 90
<210>77
<211>87
<212>PRT
<213>人乳头瘤病毒51型
<400>77
Met Tyr Leu Val Pro Ala Ala Thr Arg Tyr Pro Leu Leu Gln Leu Leu
1 5 10 15
Asn Asn Tyr Gln Thr Pro Gln Arg Pro Ile Pro Leu Pro Pro Ala Trp
20 25 30
Ala Pro Lys Lys Pro Arg His Asn Ser Glu Asn Asp Ser Asp Leu Leu
35 40 45
Ser Pro Thr Pro Pro Gln Ser Pro His Cys Pro Trp Thr Ile Gln Thr
50 55 60
Thr Lys Tyr Thr Val Glu Val Glu Ala Leu Thr Leu Glu Gly Thr Lys
65 70 75 80
Val Gln Leu Arg Leu Arg Leu
85
<210>78
<211>86
<212>PRT
<213>人乳头瘤病毒18型
<400>78
Leu Cys Ala Val Pro Val Thr Thr Arg Tyr Pro Leu Leu Ser Leu Leu
1 5 10 15
Asn Ser Tyr Ser Thr Pro Pro His Arg Ile Pro Ala Pro Cys Pro Trp
20 25 30
Ala Pro Gln Arg Pro Thr Ala Arg Arg Arg Leu Leu His Asp Leu Asp
35 40 45
Thr Val Asp Ser Arg Arg Ser Ser Ile Val Asp Leu Ser Thr His Phe
50 55 60
Ser Val Gln Leu His Leu Gln Ala Thr Thr Lys Asp Gly Asn Ser Val
65 70 75 80
Val Val Thr Leu Arg Leu
85
<210>79
<211>89
<212>PRT
<213>人乳头瘤病毒39型
<400>79
Leu Cys Ala Val Pro Val Thr Asp Arg Tyr Pro Leu Leu Asn Leu Leu
1 5 10 15
Pro Asn Tyr Gln Thr Pro Pro Arg Pro Ile Pro Pro Gln Gln Pro His
20 25 30
Ala Pro Lys Lys Gln Ser Arg Arg Arg Leu Glu Ser Asp Leu Asp Ser
35 40 45
Val Gln Ser Gln Ser Pro Leu Ser Pro Thr Glu Cys Pro Trp Thr Ile
50 55 60
Leu Thr Thr His Ser Thr Val Thr Val Gln Ala Thr Thr Gln Asp Gly
65 70 75 80
Thr Ser Val Val Val Thr Leu Arg Leu
85
<210>80
<211>94
<212>PRT
<213>人乳头瘤病毒67/68/66型
<400>80
Leu His Leu Cys Leu Val Thr Lys Tyr Pro Leu Leu Arg Leu Leu Pro
1 5 10 15
Gly Tyr His Thr Pro Gln Lys Arg Ile Pro Leu Pro Pro Pro Arg Ala
20 25 30
Pro Lys Lys Asn Arg Arg Leu Pro Asn Asp Asp Asp Leu Thr Ser Gln
35 40 45
Thr Ser Ala Thr Thr Pro Ser Thr Pro Gln Ser Tyr Cys Ala Asp Asn
50 55 60
Gly Pro Trp Thr Val His Arg Trp Gly Ser Ser Leu Asp Leu Ser Ala
65 70 75 80
Gln Thr Lys Asp Gly Val Cys Val His Leu Thr Leu His Leu
85 90
<210>81
<211>84
<212>PRT
<213>人乳头瘤病毒51型
<400>81
Leu Tyr Arg His Ile Val Thr Ile Ala Val Phe Ile Ile Leu Leu Phe
1 5 10 15
Val Leu Cys Leu Cys Val Cys Leu Val Leu Cys Cys Leu Leu Pro Leu
20 25 30
Leu Leu Ser Gln Tyr Val Phe Ala Ala Ala Leu Leu Leu Ile Leu Cys
35 40 45
Phe Trp Phe Val Val Ala Thr Ser Gln Leu Thr Thr Phe Phe Val Tyr
50 55 60
Leu Ile Phe Phe Tyr Leu Pro Cys Leu Leu Leu His Leu Tyr Thr Phe
65 70 75 80
Leu Leu Leu Gln
<210>82
<211>97
<212>PRT
<213>人乳头瘤病毒69型
<400>82
Cys Asn Ile Tyr Thr Ser His Lys Pro Ile Cys Ala Ala Asn Cys Ile
1 5 10 15
Tyr Ile Thr Ile Leu Leu Leu Val Ile Val Phe Val Leu Cys Val Cys
20 25 30
Val Cys Leu Leu Leu Cys Arg Leu Leu Pro Leu Leu Leu Ser Ile His
35 40 45
Val Phe Ala Ala His Leu Leu Ile Ile Ile Cys Phe Trp Phe Val Val
50 55 60
Ser Thr Ser Phe Thr Ala Thr Phe Phe Val Tyr Ile Cys Leu Phe Tyr
65 70 75 80
Ile Pro Ala Phe Leu Leu His Phe Tyr Ala Val Ile Leu Leu Pro Asn
85 90 95
Gly
<210>83
<211>75
<212>PRT
<213>人乳头瘤病毒52型
<400>83
Met Leu Gly Leu Phe Val Phe Cys Phe Ile Leu Leu Met Val Phe Cys
1 5 10 15
Ala Val Leu Arg Pro Leu Leu Leu Ser Ile Ser Val Tyr Ala Gln Val
20 25 30
Leu Val Leu Val Leu Leu Leu Trp Val Ser Ile Gly Ser Pro Phe Lys
35 40 45
Val Phe Phe Leu Tyr Leu Leu Phe Leu Tyr Phe Pro Met Phe Cys Ile
50 55 60
His Cys His Ala Gln Tyr Leu Ala Gln Leu Gln
65 70 75
<210>84
<211>73
<212>PRT
<213>人乳头瘤病毒67型
<400>84
Met Leu Ala Ile Phe Val Phe Ala Phe Val Leu Leu Leu Cys Phe Cys
1 5 10 15
Ile Val Leu Arg Pro Leu Leu Leu Ser Ile Tyr Val Tyr Ala Leu Leu
20 25 30
Leu Val Leu Val Leu Val Leu Trp Gly Phe Ile Gly Ser Pro Leu Arg
35 40 45
Val Phe Leu Ala Tyr Leu Ile Phe Leu Tyr Leu Pro Met Met Cys Ile
50 55 60
His Leu His Ala Gln Tyr Ile Val Ser
65 70
<210>85
<211>76
<212>PRT
<213>人乳头瘤病毒58型
<400>85
Met Ile Leu Pro Ile Phe Val Val Cys Phe Ile Leu Phe Leu Cys Leu
1 5 10 15
Cys Ile Phe Leu Arg Pro Leu Val Leu Ser Ile Ser Ile Tyr Ala Trp
20 25 30
Leu Leu Val Leu Val Leu Leu Leu Trp Val Ser Val Gly Ser Ala Leu
35 40 45
Arg Ile Phe Phe Cys Tyr Leu Ile Phe Leu Tyr Ile Pro Met Met Cys
50 55 60
Ile Asn Phe His Ala Gln Tyr Leu Thr Gln Gln Asp
65 70 75
<210>86
<211>75
<212>PRT
<213>人乳头瘤病毒33型
<400>86
Met Ile Phe Val Phe Val Leu Cys Phe Ile Leu Phe Leu Cys Leu Ser
1 5 10 15
Leu Leu Leu Arg Pro Leu Ile Leu Ser Ile Ser Thr Tyr Ala Trp Leu
20 25 30
Leu Val Leu Val Leu Leu Leu Trp Val Phe Val Gly Ser Pro Leu Lys
35 40 45
Ile Phe Phe Cys Tyr Leu Leu Phe Leu Tyr Leu Pro Met Met Cys Ile
50 55 60
Asn Phe His Ala Gln His Met Thr Gln Gln Glu
65 70 75
<210>87
<211>83
<212>PRT
<213>人乳头瘤病毒16型
<400>87
Met Thr Asn Leu Asp Thr Ala Ser Thr Thr Leu Leu Ala Cys Phe Leu
1 5 10 15
Leu Cys Phe Cys Val Leu Leu Cys Val Cys Leu Leu Ile Arg Pro Leu
20 25 30
Leu Leu Ser Val Ser Thr Tyr Thr Ser Leu Ile Ile Leu Val Leu Leu
35 40 45
Leu Trp Ile Thr Ala Ala Ser Ala Phe Arg Cys Phe Ile Val Tyr Ile
50 55 60
Ile Phe Val Tyr Ile Pro Leu Phe Leu Ile His Thr His Ala Arg Phe
65 70 75 80
Leu Ile Thr
<210>88
<211>83
<212>PRT
<213>人乳头瘤病毒35型
<400>88
Met Ile Asp Leu Thr Ala Ser Ser Thr Val Leu Leu Cys Phe Leu Leu
1 5 10 15
Cys Phe Cys Val Leu Leu Cys Leu Cys Leu Leu Val Arg Ser Leu Leu
20 25 30
Leu Ser Val Ser Leu Tyr Ser Ala Leu Ile Leu Leu Val Leu Ile Leu
35 40 45
Trp Val Thr Val Ala Thr Pro Leu Arg Cys Phe Cys Cys Phe Leu Cys
50 55 60
Phe Leu Tyr Ile Pro Met Gly Met Ile Asn Ala His Ala Gln Tyr Leu
65 70 75 80
Ala Val Gln
<210>89
<211>84
<212>PRT
<213>人乳头瘤病毒31型
<400>89
Met Ile Glu Leu Asn Ile Ser Thr Val Ser Ile Val Leu Cys Phe Leu
1 5 10 15
Leu Cys Phe Cys Val Leu Leu Phe Val Cys Leu Val Ile Arg Pro Leu
20 25 30
Val Leu Ser Val Ser Val Tyr Ala Thr Leu Leu Leu Leu Ile Val Ile
35 40 45
Leu Trp Val Ile Ala Thr Ser Pro Leu Arg Cys Phe Cys Ile Tyr Val
50 55 60
Val Phe Ile Tyr Ile Pro Leu Phe Val Ile His Thr His Ala Ser Phe
65 70 75 80
Leu Ser Gln Gln
<210>90
<211>74
<212>PRT
<213>人乳头瘤病毒73型
<400>90
Met Ile Leu Cys Ile Phe Val Phe Leu Phe Cys Val Gly Phe Cys Leu
1 5 10 15
Cys Leu Cys Val Ser Leu Ala Val Ser Val Tyr Ile Tyr Pro Trp Leu
20 25 30
Leu Val Leu Ile Ile Ile Thr Phe Ile His Val Ser Gln Ser Leu Leu
35 40 45
Lys Val Phe Phe Leu Tyr Val Leu Val Phe Tyr Ile Pro Met Ala Leu
50 55 60
Val His Tyr His Ala Thr Leu Gln Ile Thr
65 70
<210>91
<211>85
<212>PRT
<213>人乳头瘤病毒66型
<400>91
Ser Pro Tyr Ile Ala Thr Ile Asp Phe Cys Val Ile Cys Val Phe Ala
1 5 10 15
Leu Cys Phe Cys Val Cys Leu Cys Val Cys His Phe Val Pro Leu Leu
20 25 30
Leu Ser Ala Ser Leu Phe Thr Ser Cys Leu Ile Leu Ile Ile Leu Phe
35 40 45
Trp Phe Val Val Ala Thr Ser Phe Phe Asp Thr Phe Ile Leu Phe Leu
50 55 60
Leu Phe Phe Tyr Ile Pro Thr Leu Cys Ile Tyr Cys His Ala Leu Trp
65 70 75 80
Leu Ile Asn His Leu
85
<210>92
<211>73
<212>PRT
<213>人乳头瘤病毒45型
<400>92
Met Leu Ser Leu Val Phe Leu Leu Cys Phe Ser Val Cys Leu Tyr Val
1 5 10 15
Cys Cys Asn Val Pro Leu Val Gln Ser Val Tyr Val Cys Ala Phe Ala
20 25 30
Trp Leu Leu Val Phe Leu Phe Ile Val Val Ile Thr Ser Pro Leu Thr
35 40 45
Ala Phe Ala Val Tyr Ile Cys Cys Tyr Leu Leu Pro Met Phe Val Leu
50 55 60
His Met His Ala Leu His Thr Ile Gln
65 70
<210>93
<211>73
<212>PRT
<213>人乳头瘤病毒18型
<400>93
Met Leu Ser Leu Ile Phe Leu Phe Cys Phe Cys Val Cys Met Tyr Val
1 5 10 15
Cys Cys His Val Pro Leu Leu Pro Ser Val Cys Met Cys Ala Tyr Ala
20 25 30
Trp Val Leu Val Phe Val Tyr Ile Val Val Ile Thr Ser Pro Ala Thr
35 40 45
Ala Phe Thr Val Tyr Val Phe Cys Phe Leu Leu Pro Met Leu Leu Leu
50 55 60
His Ile His Ala Ile Leu Ser Leu Gln
65 70
<210>94
<211>48
<212>PRT
<213>人乳头瘤病毒68M型
<400>94
Met His Val Cys Val Tyr Val Trp Ile Leu Val Phe Val Phe Ile Leu
1 5 10 15
Val Arg Thr Thr Pro Leu Glu Val Phe Ala Val Tyr Ile Leu Phe Phe
20 25 30
Leu Leu Pro Met Trp Val Leu His Ser Phe Ala Arg Tyr Ser Met Pro
35 40 45
<210>95
<211>72
<212>PRT
<213>人乳头瘤病毒39型
<400>95
Met Ile Leu Leu Val Phe Leu Val Trp Phe Gly Val Cys Ile Tyr Ile
1 5 10 15
Cys Cys Asn Val Pro Leu Leu Pro Ser Val His Val Cys Ala Tyr Val
20 25 30
Trp Ile Ile Val Phe Val Phe Ile Leu Ile Arg Thr Thr Pro Leu Glu
35 40 45
Val Phe Phe Val Tyr Leu Leu Phe Phe Val Leu Pro Met Trp Leu Leu
50 55 60
His Arg Leu Ala Met Asp Met Ile
65 70
<210>96
<211>73
<212>PRT
<213>人乳头瘤病毒59型
<400>96
Met Ile Thr Leu Val Phe Val Cys Cys Val Cys Val Cys Leu Cys Val
1 5 10 15
Cys Cys Asn Val Pro Leu Leu Gln Ser Val Tyr Met Cys Ala Tyr Thr
20 25 30
Trp Leu Leu Val Phe Val Tyr Ile Val Val Ile Thr Ser Ser Tyr Glu
35 40 45
Cys Phe Leu Leu Tyr Ile Leu Phe Phe Ile Ile Pro Leu Leu Leu Leu
50 55 60
Tyr Ala His Ala Ile Leu Ser Ile Gln
65 70
<210>97
<211>158
<212>PRT
<213>人乳头瘤病毒16型
<400>97
Met His Gln Lys Arg Thr Ala Met Phe Gln Asp Pro Gln Glu Arg Pro
1 5 10 15
Arg Lys Leu Pro Gln Leu Cys Thr Glu Leu Gln Thr Thr Ile His Asp
20 25 30
Ile Ile Leu Glu Cys Val Tyr Cys Lys Gln Gln Leu Leu Arg Arg Glu
35 40 45
Val Tyr Asp Phe Ala Phe Arg Asp Leu Cys Ile Val Tyr Arg Asp Gly
50 55 60
Asn Pro Tyr Ala Val Cys Asp Lys Cys Leu Lys Phe Tyr Ser Lys Ile
65 70 75 80
Ser Glu Tyr Arg His Tyr Cys Tyr Ser Leu Tyr Gly Thr Thr Leu Glu
85 90 95
Gln Gln Tyr Asn Lys Pro Leu Cys Asp Leu Leu Ile Arg Cys Ile Asn
100 105 110
Cys Gln Lys Pro Leu Cys Pro Glu Glu Lys Gln Arg His Leu Asp Lys
115 120 125
Lys Gln Arg Phe His Asn Ile Arg Gly Arg Trp Thr Gly Arg Cys Met
130 135 140
Ser Cys Cys Arg Ser Ser Arg Thr Arg Arg Glu Thr Gln Leu
145 150 155
<210>98
<211>160
<212>PRT
<213>人乳头瘤病毒59型
<400>98
Met Ala Arg Phe Glu Asp Pro Thr Gln Arg Pro Tyr Lys Leu Pro Asp
1 5 10 15
Leu Ser Thr Thr Leu Asn Ile Pro Leu His Asp Ile Arg Ile Asn Cys
20 25 30
Val Phe Cys Lys Gly Glu Leu Gln Glu Arg Glu Val Phe Glu Phe Ala
35 40 45
Phe Asn Asp Leu Phe Ile Val Tyr Arg Asp Cys Thr Pro Tyr Ala Ala
50 55 60
Cys Leu Lys Cys Ile Ser Phe Tyr Ala Arg Val Arg Glu Leu Arg Tyr
65 70 75 80
Tyr Arg Asp Ser Val Tyr Gly Glu Thr Leu Glu Ala Glu Thr Lys Thr
85 90 95
Pro Leu His Glu Leu Leu Ile Arg Cys Tyr Arg Cys Leu Lys Pro Leu
100 105 110
Cys Pro Thr Asp Lys Leu Lys His Ile Thr Glu Lys Arg Arg Phe His
115 120 125
Asn Ile Ala Gly Ile Tyr Thr Gly Gln Cys Arg Gly Cys Arg Thr Arg
130 135 140
Ala Arg His Leu Arg Gln Gln Arg Gln Ala Arg Ser Glu Thr Leu Val
145 150 155 160
<210>99
<211>158
<212>PRT
<213>人乳头瘤病毒39型
<400>99
Met Ala Arg Phe His Asn Pro Ala Glu Arg Pro Tyr Lys Leu Pro Asp
1 5 10 15
Leu Cys Thr Thr Leu Asp Thr Thr Leu Gln Asp Ile Thr Ile Ala Cys
20 25 30
Val Tyr Cys Arg Arg Pro Leu Gln Gln Thr Glu Val Tyr Glu Phe Ala
35 40 45
Phe Ser Asp Leu Tyr Val Val Tyr Arg Asp Gly Glu Pro Leu Ala Ala
50 55 60
Cys Gln Ser Cys Ile Lys Phe Tyr Ala Lys Ile Arg Glu Leu Arg Tyr
65 70 75 80
Tyr Ser Asp Ser Val Tyr Ala Thr Thr Leu Glu Asn Ile Thr Asn Thr
85 90 95
Lys Leu Tyr Asn Leu Leu Ile Arg Cys Met Cys Cys Leu Lys Pro Leu
100 105 110
Cys Pro Ala Glu Lys Leu Arg His Leu Asn Ser Lys Arg Arg Phe His
115 120 125
Lys Ile Ala Gly Ser Tyr Thr Gly Gln Cys Arg Arg Cys Trp Thr Thr
130 135 140
Lys Arg Glu Asp Arg Arg Leu Thr Arg Arg Glu Thr Gln Val
145 150 155
<210>100
<211>158
<212>PRT
<213>人乳头瘤病毒45型
<400>100
Met Ala Arg Phe Asp Asp Pro Lys Gln Arg Pro Tyr Lys Leu Pro Asp
1 5 10 15
Leu Cys Thr Glu Leu Asn Thr Ser Leu Gln Asp Val Ser Ile Ala Cys
20 25 30
Val Tyr Cys Lys Ala Thr Leu Glu Arg Thr Glu Val Tyr Gln Phe Ala
35 40 45
Phe Lys Asp Leu Cys Ile Val Tyr Arg Asp Cys Ile Ala Tyr Ala Ala
50 55 60
Cys His Lys Cys Ile Asp Phe Tyr Ser Arg Ile Arg Glu Leu Arg Tyr
65 70 75 80
Tyr Ser Asn Ser Val Tyr Gly Glu Thr Leu Glu Lys Ile Thr Asn Thr
85 90 95
Glu Leu Tyr Asn Leu Leu Ile Arg Cys Leu Arg Cys Gln Lys Pro Leu
100 105 110
Asn Pro Ala Glu Lys Arg Arg His Leu Lys Asp Lys Arg Arg Phe His
115 120 125
Ser Ile Ala Gly Gln Tyr Arg Gly Gln Cys Asn Thr Cys Cys Asp Gln
130 135 140
Ala Arg Gln Glu Arg Leu Arg Arg Arg Arg Glu Thr Gln Val
145 150 155
<210>101
<211>154
<212>PRT
<213>人乳头瘤病毒66型
<400>101
Met Asp Ser Ile Phe Ser Asn Thr Gln Glu Arg Pro Arg Ser Leu His
1 5 10 15
His Leu Ser Glu Val Leu Gln Ile Pro Leu Leu Asp Leu Arg Leu Ser
20 25 30
Cys Val Tyr Cys Lys Lys Glu Leu Thr Ser Leu Glu Leu Tyr Arg Phe
35 40 45
Ala Cys Ile Glu Leu Lys Leu Val TyrArg Asn Asn Trp Pro Tyr Ala
50 55 60
Val Cys Arg Val Cys Leu Leu Phe Tyr Ser Lys Val Arg Lys Tyr Arg
65 70 75 80
Tyr Tyr Lys Tyr Ser Val Tyr Gly Ala Thr Leu Glu Ser Ile Thr Lys
85 90 95
Lys Gln Leu Ser Asp Leu Ser Ile Arg Cys Tyr Arg Cys Gln Cys Pro
100 105 110
Leu Thr Pro Glu Glu Lys Gln Leu His Cys Glu His Lys Arg Arg Phe
115 120 125
His Tyr Ile Ala Tyr Ala Trp Thr Gly Ser Cys Leu Gln Cys Trp Arg
130 135 140
His Thr Ser Arg Gln Ala Thr Glu Ser Val
145 150
<210>102
<211>150
<212>PRT
<213>人乳头瘤病毒26型
<400>102
Met Phe Glu Asp Pro Arg Glu Arg Pro Arg Thr Leu His Glu Leu Cys
1 5 10 15
Glu Ser Leu Asn Thr Thr Leu Gln Asn Leu Gln Val Gln Cys Val Tyr
20 25 30
Cys Lys Glu Thr Leu Gln Trp Ala Asp Val Tyr Asn Phe Ala Ile Cys
35 40 45
Asp Leu Arg Val Val Tyr Arg Asp Arg Ser Pro Tyr Ala Ala Cys Lys
50 55 60
Arg Cys Val Ile Phe Tyr Ser Lys Ile Thr Glu Tyr Arg Arg Tyr Thr
65 70 75 80
Cys Ser Val Tyr Gly Ala Thr Leu Glu Ala Leu Thr Lys Lys Ser Leu
85 90 95
Cys Asn Leu Leu Ile Arg Cys His Arg Cys Gln Met Pro Leu Gly Pro
100 105 110
Glu Glu Lys Gln Arg Ile Val Asp Glu Lys Arg Arg Phe His Glu Ile
115 120 125
Ala Gly Gln Trp Lys Gly Leu Cys Thr Asn Cys Trp Arg Pro Arg Arg
130 135 140
Gln Thr Glu Thr Gln Val
145 150
<210>103
<211>151
<212>PRT
<213>人乳头瘤病毒51型
<400>103
Met Phe Glu Asp Lys Arg Glu Arg Pro Arg Thr Leu His Glu Leu Cys
1 5 10 15
Glu Ala Leu Asn Val Ser Met His Asn Ile Gln Val Val Cys Val Tyr
20 25 30
Cys Lys Lys Glu Leu Cys Arg Ala Asp Val Tyr Asn Val Ala Phe Thr
35 40 45
Glu Ile Lys Ile Val Tyr Arg Asp Asn Asn Pro Tyr Ala Val Cys Lys
50 55 60
Gln Cys Leu Leu Phe Tyr Ser Lys Ile Arg Glu Tyr ArgArg Tyr Ser
65 70 75 80
Arg Ser Val Tyr Gly Thr Thr Leu Glu Ala Ile Thr Lys Lys Ser Leu
85 90 95
Tyr Asp Leu Ser Ile Arg Cys His Arg Cys Gln Arg Pro Leu Gly Pro
100 105 110
Glu Glu Lys Gln Lys Leu Val Asp Glu Lys Lys Arg Phe His Glu Ile
115 120 125
Ala Gly Arg Trp Thr Gly Gln Cys Ala Asn Cys Trp Gln Arg Thr Arg
130 135 140
Gln Arg Asn Glu Thr Gln Val
145 150
<210>104
<211>151
<212>PRT
<213>人乳头瘤病毒82型
<400>104
Met Phe Glu Asp Ile Arg Glu Arg Pro Arg Thr Leu His Glu Leu Cys
1 5 10 15
Glu Ala Cys Asn Thr Ser Met His Asn Ile Gln Val Leu Cys Val Tyr
20 25 30
Cys Lys Lys Glu Leu Cys Arg Ala Asp Val Tyr Asn Val Ala Phe Thr
35 40 45
Glu Leu Arg Ile Val Tyr Arg Asp Asn Thr Pro Tyr Ala Ala Cys Lys
50 55 60
Lys Cys Leu Met Phe Tyr Ser Arg Ile Arg Glu Tyr Arg Arg Tyr Ser
65 70 75 80
Arg Ser Val Tyr Gly Ala Thr Leu Glu Ala Ile Thr Asn Lys Ser Leu
85 90 95
Tyr Glu Leu Leu Ile Arg Cys His Arg Cys Gln Arg Pro Leu Gly Pro
100 105 110
Glu Glu Lys Gln Lys Val Val Asp Asp Lys Lys Arg Phe His Glu Ile
115 120 125
Ala Gly Arg Trp Thr Gly Gln Cys Ala Asn Cys Arg Lys Pro Pro Arg
130 135 140
Gln Arg Ser Glu Thr Gln Val
145 150
<210>105
<211>146
<212>PRT
<213>人乳头瘤病毒52型
<400>105
Met Phe Glu Asp Pro Ala Thr Arg Pro Arg Thr Leu His Glu Leu Cys
1 5 10 15
Glu Val Leu Glu Glu Ser Val His Glu Ile Arg Leu Gln Cys Val Gln
20 25 30
Cys Lys Lys Glu Leu Gln Arg Arg Glu Val Tyr Lys Phe Leu Phe Thr
35 40 45
Asp Leu Arg Ile Val Tyr Arg Asp Asn Asn Pro Tyr Gly Val Cys Ile
50 55 60
Met Cys Leu Arg Phe Leu Ser Lys Ile Ser Glu Tyr Arg His Tyr Gln
65 70 75 80
Tyr Ser Leu Tyr Gly Lys Thr Leu Glu Glu Arg Val Lys Lys Pro Leu
85 90 95
Ser Glu Ile Thr Ile Arg Cys Ile Ile Cys Gln Thr Pro Leu Cys Pro
100 105 110
Glu Glu Lys Glu Arg His Val Asn Ala Asn Lys Arg Phe His Asn Ile
115 120 125
Met Gly Arg Trp Thr Gly Arg Cys Ser Glu Cys Trp Arg Pro Arg Pro
130 135 140
Val Thr
145
<210>106
<211>149
<212>PRT
<213>人乳头瘤病毒58型
<400>106
Met Phe Gln Asp Ala Glu Glu Lys Pro Arg Thr Leu His Asp Leu Cys
1 5 10 15
Gln Ala Leu Glu Thr Ser Val His Glu Ile Glu Leu Lys Cys Val Glu
20 25 30
Cys Lys Lys Thr Leu Gln Arg Ser Glu Val Tyr Asp Phe Val Phe Ala
35 40 45
Asp Leu Arg Ile Val Tyr Arg Asp Gly Asn Pro Phe Ala Val Cys Lys
50 55 60
Val Cys Leu Arg Leu Leu Ser Lys Ile Ser Glu Tyr Arg His Tyr Asn
65 70 75 80
Tyr Ser Leu Tyr Gly Asp Thr Leu Glu Gln Thr Leu Lys Lys Cys Leu
85 90 95
Asn Glu Ile Leu Ile Arg Cys Ile Ile Cys Gln Arg Pro Leu Cys Pro
100 105 110
Gln Glu Lys Lys Arg His Val Asp Leu Asn Lys Arg Phe His Asn Ile
115 120 125
Ser Gly Arg Trp Thr Gly Arg Cys Ala Val Cys Trp Arg Pro Arg Arg
130 135 140
Arg Gln Thr Gln Val
145
<210>107
<211>149
<212>PRT
<213>人乳头瘤病毒33型
<400>107
Met Phe Gln Asp Thr Glu Glu Lys Pro Arg Thr Leu His Asp Leu Cys
1 5 10 15
Gln Ala Leu Glu Thr Thr Ile His Asn Ile Glu Leu Gln Cys Val Glu
20 25 30
Cys Lys Lys Pro Leu Gln Arg Ser Glu Val Tyr Asp Phe Ala Phe Ala
35 40 45
Asp Leu Thr Val Val Tyr Arg Glu Gly Asn Pro Phe Gly Ile Cys Lys
50 55 60
Leu Cys Leu Arg Phe Leu Ser Lys Ile Ser Glu Tyr Arg His Tyr Asn
65 70 75 80
Tyr Ser Val Tyr Gly Asn Thr Leu Glu Gln Thr Val Lys Lys Pro Leu
85 90 95
Asn Glu Ile Leu Ile Arg Cys Ile Ile Cys Gln Arg Pro Leu Cys Pro
100 105 110
Gln Glu Lys Lys Arg His Val Asp Leu Asn Lys Arg Phe His Asn Ile
115 120 125
Ser Gly Arg Trp Ala Gly Arg Cys Ala Ala Cys Trp Arg Ser Arg Arg
130 135 140
Arg Glu Thr Ala Leu
145
<210>108
<211>149
<212>PRT
<213>人乳头瘤病毒31型
<400>108
Met Phe Lys Asn Pro Ala Glu Arg Pro Arg Lys Leu His Glu Leu Ser
1 5 10 15
Ser Ala Leu Glu Ile Pro Tyr Asp Glu Leu Arg Leu Asn Cys Val Tyr
20 25 30
Cys Lys Gly Gln Leu Thr Glu Thr Glu Val Leu Asp Phe Ala Phe Thr
35 40 45
Asp Leu Thr Ile Val Tyr Arg Asp Asp Thr Pro His Gly Val Cys Thr
50 55 60
Lys Cys Leu Arg Phe Tyr Ser Lys Val Ser Glu Phe Arg Trp Tyr Arg
65 70 75 80
Tyr Ser Val Tyr Gly Thr Thr Leu Glu Lys Leu Thr Asn Lys Gly Ile
85 90 95
Cys Asp Leu Leu Ile Arg Cys Ile Thr Cys Gln Arg Pro Leu Cys Pro
100 105 110
Glu Glu Lys Gln Arg His Leu Asp Lys Lys Lys Arg Phe His Asn Ile
115 120 125
Gly Gly Arg Trp Thr Gly Arg Cys Ile Ala Cys Trp Arg Arg Pro Arg
130 135 140
Thr Glu Thr Gln Val
145
<210>109
<211>149
<212>PRT
<213>人乳头瘤病毒35型
<400>109
Met Phe Gln Asp Pro Ala Glu Arg Pro Tyr Lys Leu His Asp Leu Cys
1 5 10 15
Asn Glu Val Glu Glu Ser Ile His Glu Ile Cys Leu Asn Cys Val Tyr
20 25 30
Cys Lys Gln Glu Leu Gln Arg Ser Glu Val Tyr Asp Phe Ala Cys Tyr
35 40 45
Asp Leu Cys Ile Val Tyr Arg Glu Gly Gln Pro Tyr Gly Val Cys Met
50 55 60
Lys Cys Leu Lys Phe Tyr Ser Lys Ile Ser Glu Tyr Arg Trp Tyr Arg
65 70 75 80
Tyr Ser Val Tyr Gly Glu Thr Leu Glu Lys Gln Cys Asn Lys Gln Leu
85 90 95
Cys His Leu Leu Ile Arg Cys Ile Thr Cys Gln Lys Pro Leu Cys Pro
100 105 110
Val Glu Lys Gln Arg His Leu Glu Glu Lys Lys Arg Phe His Asn Ile
115 120 125
Gly Gly Arg Trp Thr Gly Arg Cys Met Ser Cys Trp Lys Pro Thr Arg
130 135 140
Arg Glu Thr Glu Val
145
<210>110
<211>158
<212>PRT
<213>人乳头瘤病毒18型
<400>110
Met Ala Arg Phe Glu Asp Pro Thr Arg Arg Pro Tyr Lys Leu Pro Asp
1 5 10 15
Leu Cys Thr Glu Leu Asn Thr Ser Leu Gln Asp Ile Glu Ile Thr Cys
20 25 30
Val Tyr Cys Lys Thr Val Leu Glu Leu Thr Glu Val Phe Glu Phe Ala
35 40 45
Phe Lys Asp Leu Phe Val Val Tyr Arg Asp Ser Ile Pro His Ala Ala
50 55 60
Cys His Lys Cys Ile Asp Phe Tyr Ser Arg Ile Arg Glu Leu Arg His
65 70 75 80
Tyr Ser Asp Ser Val Tyr Gly Asp Thr Leu Glu Lys Leu Thr Asn Thr
85 90 95
Gly Leu Tyr Asn Leu Leu Ile Arg Cys Leu Arg Cys Gln Lys Pro Leu
100 105 110
Asn Pro Ala Glu Lys Leu Arg His Leu Asn Glu Lys Arg Arg Phe His
115 120 125
Asn Ile Ala Gly His Tyr Arg Gly Gln Cys His Ser Cys Cys Asn Arg
130 135 140
Ala Arg Gln Glu Arg Leu Gln Arg Arg Arg Glu Thr Gln Val
145 150 155
<210>111
<211>98
<212>PRT
<213>人乳头瘤病毒16型
<400>111
Met His Gly Asp Thr Pro Thr Leu His Glu Tyr Met Leu Asp Leu Gln
1 5 10 15
Pro Glu Thr Thr Asp Leu Tyr Cys Tyr Glu Gln Leu Asn Asp Ser Ser
20 25 30
Glu Glu Glu Asp Glu Ile Asp Gly Pro Ala Gly Gln Ala Glu Pro Asp
35 40 45
Arg Ala His Tyr Asn Ile Val Thr Phe Cys Cys Lys Cys Asp Ser Thr
50 55 60
Leu Arg Leu Cys Val Gln Ser Thr His Val Asp Ile Arg Thr Leu Glu
65 70 75 80
Asp Leu Leu Met Gly Thr Leu Gly Ile Val Cys Pro Ile Cys Ser Gln
85 90 95
Lys Pro
<210>112
<211>104
<212>PRT
<213>人乳头瘤病毒18型
<400>112
Met His Gly Pro Lys Ala Thr Leu Gln Asp Ile Val Leu His Leu Glu
1 5 10 15
Pro Gln Asn Glu Ile Pro Val Asp Leu Leu Cys His Glu Gln Leu Ser
20 25 30
Asp Ser Glu Glu Glu Asn Asp Glu Ile Asp Gly Val Asn His Gln His
35 40 45
Leu Pro Ala Arg Arg Ala Glu Pro Gln Arg His Met Leu Cys Met Cys
50 55 60
Cys Lys Cys Glu Ala Arg Ile Lys Leu Val Val Glu Ser Ser Ala Asp
65 70 75 80
Asp Leu Arg Ala Phe Gln Gln Leu Phe Leu Asn Thr Leu Ser Phe Val
85 90 95
Cys Pro Trp Cys Ala Ser Gln Gln
100
<210>113
<211>98
<212>PRT
<213>人乳头瘤病毒31型
<400>113
Met Arg Gly Glu Thr Pro Thr Leu Gln Asp Tyr Val Leu Asp Leu Gln
1 5 10 15
Pro Lys Ala Thr Asp Leu His Cys Tyr Glu Gln Leu Pro Asp Ser Ser
20 25 30
Asp Glu Glu Asp Val Ile Asp Ser Pro Ala Gly Gln Ala Lys Pro Asp
35 40 45
Thr Ser Asn Tyr Asn Ile Val Thr Phe Cys Cys Gln Cys Glu Ser Thr
50 55 60
Leu Arg Leu Cys Val Gln Ser Thr Gln Val Asp Ile Arg Ile Leu Gln
65 70 75 80
Glu Leu Leu Met Gly Ser Phe Gly Ile Val Cys Pro Asn Cys Ser Thr
85 90 95
Arg Leu
<210>114
<211>97
<212>PRT
<213>人乳头瘤病毒33型
<400>114
Met Arg Gly His Lys Pro Thr Leu Lys Glu Tyr Val Leu Asp Leu Tyr
1 5 10 15
Pro Glu Pro Thr Asp Leu Tyr Cys Tyr Glu Gln Leu Ser Asp Ser Ser
20 25 30
Asp Glu Asp Glu Gly Leu Asp Arg Pro Asp Gly Gln Ala Gln Pro Ala
35 40 45
Thr Ala Asp Tyr Tyr Ile Val Thr Cys Cys His Thr Cys Asn Thr Thr
50 55 60
Val Arg Leu Cys Val Asn Ser Thr Ala Ser Asp Leu Arg Thr Ile Gln
65 70 75 80
Gln Leu Leu Met Gly Thr Val Asn Ile Val Cys Pro Thr Cys Ala Gln
85 90 95
Gln
<210>115
<211>99
<212>PRT
<213>人乳头瘤病毒35型
<400>115
Met His Gly Glu Ile Thr Thr Leu Gln Asp Tyr Val Leu Asp Leu Glu
1 5 10 15
Pro Glu Ala Thr Asp Leu Tyr Cys Tyr Glu Gln Leu Cys Asp Ser Ser
20 25 30
Glu Glu Glu Glu Asp Thr Ile Asp Gly Pro Ala Gly Gln Ala Lys Pro
35 40 45
Asp Thr Ser Asn Tyr Asn Ile Val Thr Ser Cys Cys Lys Cys Glu Ala
50 55 60
Thr Leu Arg Leu Cys Val Gln Ser Thr His Ile Asp Ile Arg Lys Leu
65 70 75 80
Glu Asp Leu Leu Met Gly Thr Phe Gly Ile Val Cys Pro Gly Cys Ser
85 90 95
Gln Arg Ala
<210>116
<211>102
<212>PRT
<213>人乳头瘤病毒39型
<400>116
Met Arg Gly Pro Lys Pro Thr Leu Gln Glu Ile Val Leu Asp Leu Cys
1 5 10 15
Pro Tyr Asn Glu Ile Gln Pro Val Asp Leu Val Cys His Glu Gln Leu
20 25 30
Gly Glu Ser Glu Asp Glu Ile Asp Glu Pro Asp His Ala Val Asn His
35 40 45
Gln His Gln Leu Leu Ala Arg Arg Asp Glu Pro Gln Arg His Thr Ile
50 55 60
Gln Cys Ser Cys Cys Lys Cys Asn Asn Thr Leu Gln Leu Val Val Glu
65 70 75 80
Ala Ser Arg Asp Thr Leu Arg Gln Leu Gln Gln Leu Phe Met Asp Ser
85 90 95
Leu Gly Phe Val Cys Pro
100
<210>117
<211>102
<212>PRT
<213>人乳头瘤病毒45型
<400>117
Met His Gly Pro Arg Glu Thr Leu Gln Glu Ile Val Leu His Leu Glu
1 5 10 15
Pro Gln Asn Glu Leu Asp Pro Val Asp Leu Leu Cys Tyr Glu Gln Leu
20 25 30
Ser Glu Ser Glu Glu Glu Asn Asp Glu Ala Asp Gly Val Ser His Ala
35 40 45
Gln Leu Pro Ala Arg Arg Ala Glu Pro Gln Arg His Lys Ile Leu Cys
50 55 60
Val Cys Cys Lys Cys Asp Gly Arg Ile Glu Leu Thr Val Glu Ser Ser
65 70 75 80
Ala Glu Asp Leu Arg Thr Leu Gln Gln Leu Phe Leu Ser Thr Leu Ser
85 90 95
Phe Val Cys Pro Trp Cys
100
<210>118
<211>101
<212>PRT
<213>人乳头瘤病毒51型
<400>118
Met Arg Gly Asn Val Pro Gln Leu Lys Asp Val Val Leu His Leu Thr
1 5 10 15
Pro Gln Thr Glu Ile Asp Leu Gln Cys Tyr Glu Gln Phe Asp Ser Ser
20 25 30
Glu Glu Glu Asp Glu Val Asp Asn Met Arg Asp Gln Leu Pro Glu Arg
35 40 45
Arg Ala Gly Gln Ala Thr Cys Tyr Arg Ile Glu Ala Pro Cys Cys Arg
50 55 60
Cys Ser Ser Val Val Gln Leu Ala Val Glu Ser Ser Gly Asp Thr Leu
65 70 75 80
Arg Val Val Gln Gln Met Leu Met Gly Glu Leu Ser Leu Val Cys Pro
85 90 95
Cys Cys Ala Asn Asn
100
<210>119
<211>99
<212>PRT
<213>人乳头瘤病毒52型
<400>119
Met Arg Gly Asp Lys Ala Thr Ile Lys Asp Tyr Ile Leu Asp Leu Gln
1 5 10 15
Pro Glu Thr Thr Asp Leu His Cys Tyr Glu Gln Leu Gly Asp Ser Ser
20 25 30
Asp Glu Glu Asp Thr Asp Gly Val Asp Arg Pro Asp Gly Gln Ala Glu
35 40 45
Gln Ala Thr Ser Ash Tyr Tyr Ile Val Thr Tyr Cys His Ser Cys Asp
50 55 60
Ser Thr Leu Arg Leu Cys Ile His Ser Thr Ala Thr Asp Leu Arg Thr
65 70 75 80
Leu Gln Gln Met Leu Leu Gly Thr Leu Gln Val Val Cys Pro Gly Cys
85 90 95
Ala Arg Leu
<210>120
<211>104
<212>PRT
<213>人乳头瘤病毒66型
<400>120
Met His Gly Lys Val Pro Thr Leu Gln Glu Val Ile Leu Glu Leu Ala
1 5 10 15
Pro Gln Thr Glu Ile Asp Leu Gln Cys Asn Glu Gln Leu Asp Ser Ser
20 25 30
Glu Asp Glu Asp Glu Asp Glu Ile Asp His Leu Leu Glu Arg Pro Gln
35 40 45
Gln Ala Arg Gln Ala Glu Gln His Lys Cys Tyr Leu Ile His Val Pro
50 55 60
Cys Cys Lys Cys Glu Leu Val Val Gln Leu Asp Ile Gln Ser Thr Lys
65 70 75 80
Glu Glu Leu Arg Val Val Gln Gln Leu Leu Met Gly Ala Leu Thr Val
85 90 95
Thr Cys Pro Leu Cys Ala Ser Lys
100
<210>121
<211>110
<212>PRT
<213>人乳头瘤病毒68型
<400>121
Met His Gly Pro Lys Pro Thr Val Gln Glu Ile Val Leu Glu Leu Cys
1 5 10 15
Pro Tyr Asn Glu Ile Gln Pro Val Asp Leu Val Cys His Glu Gln Leu
20 25 30
Gly Asp Ser Asp Asp Glu Ile Asp Glu Pro Asp His Ala Val Asn His
35 40 45
His Gln His Leu Leu Leu Ala Arg Arg Asp Glu Gln Gln Arg His Arg
50 55 60
Ile Gln Cys Leu Cys Cys Lys Cys Asn Lys Ala Leu Gln Leu Val Val
65 70 75 80
Glu Ala Ser Arg Asp Asn Leu Arg Thr Leu Gln Gln Leu Phe Met Asp
85 90 95
Ser Leu Asn Phe Val Cys Pro Trp Cys Ala Thr Glu Thr Gln
100 105 110
<210>122
<211>92
<212>PRT
<213>人乳头瘤病毒56/57/58型
<400>122
Met His Gly Glu Arg Pro Ser Leu Glu Asp Ile Thr Leu Ile Leu Glu
1 5 10 15
Glu Ile Pro Glu Ile Val Asp Leu His Cys Asp Glu Gln Phe Asp Asn
20 25 30
Ser Glu Glu Asp Thr Asn Tyr Gln Leu Thr Glu Pro Ala Val Gln Ala
35 40 45
Tyr Gly Val Val Thr Thr Cys Cys Lys Cys His Ser Thr Val Arg Leu
50 55 60
Val Val Glu Cys Gly Ala Ala Asp Ile Arg His Leu Glu Gln Leu Phe
65 70 75 80
Leu Asn Thr Leu Thr Ile Val Cys Pro Arg Cys Val
85 90
Claims (17)
1.一种筛查人乳头瘤病毒(HPV)整合的方法，该方法包括：
(a)将HPV整合检测指示物施加给细胞和细胞群，所述细胞和细胞群是得自宫颈的细胞和细胞群样品的一部分；
(b)人工或采用自动仪器读取通过分析细胞和细胞群获得的信息。
2.如权利要求1所述的方法，其特征在于，所述HPV整合检测指示物是抗体。
3.如权利要求2所述的方法，其特征在于，所述抗体是抗HPV整合指示物的特异性抗体。
4.如权利要求3所述的方法，其特征在于，所述HPV整合指示物是E1、E4、E5、E6和E7蛋白中的至少一种。
5.如权利要求2所述的方法，其特征在于，所述抗体与含有共有氨基酸序列的多肽反应，从而所述抗体与多于一种的高风险HPV类型的多肽反应。
6.如权利要求2所述的方法，其特征在于，所述抗体针对一多肽而产生，该多肽含有SEQ ID NO：1、SEQ ID NO：2、SEQ ID NO：3、SEQ ID NO：4、SEQID NO：5、SEQ ID NO：6、SEQ ID NO：7、SEQ ID NO：8、SEQ ID NO：9、SEQID NO：10和SEQ ID NO：11中的至少一条的氨基酸序列。
7.如权利要求2所述的方法，其特征在于，所述抗体针对一多肽而产生，该多肽含有SEQ ID NO：12、SEQ ID NO：13和SEQ ID NO：14中的至少一条的氨基酸序列。
8.如权利要求2所述的方法，其特征在于，所述抗体针对一多肽而产生，该多肽含有SEQ ID NO：15、SEQ ID NO：16、SEQ ID NO：17、SEQ ID NO：18、SEQ ID NO：19和SEQ ID NO：20中的至少一条的氨基酸序列。
9.如权利要求2所述的方法，其特征在于，所述抗体针对一多肽而产生，该多肽含有SEQ ID NO：21、SEQ ID NO：22、SEQ ID NO：23、SEQ ID NO：24、SEQ ID NO：25和SEQ ID NO：26中的至少一条的氨基酸序列。
10.如权利要求2所述的方法，其特征在于，所述抗体针对一多肽而产生，该多肽含有SEQ ID NO：27、SEQ ID NO：28和SEQ ID NO：29中的至少一条的氨基酸序列。
11.一种检测宫颈中的异常组织的方法，该方法包括：
使用采样器从宫颈采集细胞和细胞群；
将采样器采集到的细胞和细胞群的至少一些转移到接受结构或转换成悬浮形式的液体基样品，该液体基样品可被转移到该接受结构；
将细胞学人乳头瘤病毒(HPV)整合检测指示物施加到被转移到该接受结构的细胞和细胞群中；和
人工或采用自动仪器分析所述细胞和细胞群的HPV整合。
12.如权利要求11所述的方法，其特征在于，所述方法还包括在将所述细胞和细胞群采集到小瓶中的悬浮液中、并将所述细胞和细胞群转移到所述接受结构后将剩余的细胞放置在采样器上的步骤。
13.如权利要求12所述的方法，其特征在于，所述小瓶中等份的采集细胞用于免疫学检测HPV整合。
14.如权利要求11所述的方法，其特征在于，将所有采集的细胞置入小瓶或其它容器的悬浮液中。
15.如权利要求14所述的方法，其特征在于，所述小瓶中的悬浮细胞用于免疫学检测HPV整合。
16.如权利要求11所述的方法，其特征在于，所述接受结构是载玻片、培养皿或膜。
17.如权利要求11所述的方法，其特征在于，所述方法还包括：
根据巴帕尼科拉乌方法给转移到接受结构的细胞和细胞群染色，并根据贝色斯达系统评价染色的细胞。
Applications Claiming Priority (7)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| US97506907P | 2007-09-25 | 2007-09-25 | |
| US60/975,069 | 2007-09-25 | ||
| US99289207P | 2007-12-06 | 2007-12-06 | |
| US60/992,892 | 2007-12-06 | ||
| US12/211,547 US7838215B2 (en) | 2007-09-25 | 2008-09-16 | Advanced cervical cell screening methods |
| US12/211,547 | 2008-09-16 | ||
| PCT/US2008/076813 WO2009042488A2 (en) | 2007-09-25 | 2008-09-18 | Advanced cervical cell screening methods |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN101932711A true CN101932711A (zh) | 2010-12-29 |
Family
ID=40512084
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN2008801093612A Pending CN101932711A (zh) | 2007-09-25 | 2008-09-18 | 晚期宫颈细胞筛查方法 |
Country Status (6)
| Country | Link |
|---|---|
| US (1) | US7838215B2 (zh) |
| EP (1) | EP2201119A4 (zh) |
| JP (1) | JP2010540932A (zh) |
| CN (1) | CN101932711A (zh) |
| CA (1) | CA2700635A1 (zh) |
| WO (1) | WO2009042488A2 (zh) |
Families Citing this family (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US8968995B2 (en) * | 2008-11-12 | 2015-03-03 | Oncohealth Corp. | Detection, screening, and diagnosis of HPV-associated cancers |
| US8278056B2 (en) * | 2008-06-13 | 2012-10-02 | Oncohealth Corp. | Detection of early stages and late stages HPV infection |
| EP2427763A4 (en) | 2009-05-07 | 2013-08-21 | Oncohealth Corp | IDENTIFICATION OF A HIGH CIN2 DEGREE FOR THE DETECTION OF EARLY AND LATE TADIES AND FOR THE SCREENING AND DIAGNOSIS OF HUMAN PAPILLOMA (HPV) AND ASSOCIATED CANCER DISORDERS |
| WO2011084598A1 (en) | 2010-01-08 | 2011-07-14 | Oncohealth Corporation | High throughput cell-based hpv immunoassays for diagnosis and screening of hpv-associated cancers |
| US20120264110A1 (en) * | 2011-04-12 | 2012-10-18 | Wachman Elliot S | Automated pap screening using a plurality of biomarkers and multi-spectral imaging |
| DE102011053741A1 (de) * | 2011-09-19 | 2013-03-21 | Ralf Hilfrich | Verfahren zum Überprüfen der Selbstheilung durch das Immunsystem eines mit humanen Papillomviren infizierten Menschen |
| CN104830869B (zh) * | 2015-04-30 | 2018-06-01 | 武汉凯德维斯生物技术有限公司 | 与宫颈癌发生相关的hpv整合的基因位点及其应用 |
Family Cites Families (10)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| GB9718745D0 (en) * | 1996-12-03 | 1997-11-12 | Medical Res Council | Improvements in or relating to screening for carcinoma |
| US6183746B1 (en) * | 1997-10-09 | 2001-02-06 | Zycos Inc. | Immunogenic peptides from the HPV E7 protein |
| GB0018140D0 (en) * | 2000-07-24 | 2000-09-13 | Medical Res Council | Screening for abnormalities |
| US6884605B2 (en) * | 2001-08-09 | 2005-04-26 | Board Of Trustees Of The University Of Arkansas | Compositions, methods and products comprising human papillomavirus for detecting and treating a cancer |
| EA006975B1 (ru) * | 2002-01-07 | 2006-06-30 | Норчип А/С | Способ обнаружения мрнк вируса папилломы человека |
| EP1369694A1 (en) | 2002-04-09 | 2003-12-10 | MTM Laboratories AG | Method for discrimination of metaplasias from neoplastic or preneoplastic lesions |
| JP5030594B2 (ja) * | 2003-12-23 | 2012-09-19 | アルボー ビータ コーポレーション | Ｈｐｖの発癌性株に対する抗体およびそれらの使用方法 |
| ATE549625T1 (de) * | 2004-03-24 | 2012-03-15 | Tripath Imaging Inc | Verfahren und zusammensetzungen zum nachweis einer zervixerkrankung |
| US7482142B1 (en) * | 2004-05-07 | 2009-01-27 | Roche Molecular Systems, Inc. | High-risk human papillomavirus detection |
| US20060189893A1 (en) * | 2005-01-06 | 2006-08-24 | Diamics, Inc. | Systems and methods for detecting abnormal cells |
-
2008
- 2008-09-16 US US12/211,547 patent/US7838215B2/en not_active Expired - Fee Related
- 2008-09-18 CN CN2008801093612A patent/CN101932711A/zh active Pending
- 2008-09-18 CA CA2700635A patent/CA2700635A1/en not_active Abandoned
- 2008-09-18 EP EP08834679A patent/EP2201119A4/en not_active Withdrawn
- 2008-09-18 WO PCT/US2008/076813 patent/WO2009042488A2/en not_active Ceased
- 2008-09-18 JP JP2010527038A patent/JP2010540932A/ja not_active Withdrawn
Also Published As
| Publication number | Publication date |
|---|---|
| US7838215B2 (en) | 2010-11-23 |
| US20090104597A1 (en) | 2009-04-23 |
| WO2009042488A2 (en) | 2009-04-02 |
| CA2700635A1 (en) | 2009-04-02 |
| EP2201119A4 (en) | 2010-11-24 |
| WO2009042488A3 (en) | 2009-05-22 |
| EP2201119A2 (en) | 2010-06-30 |
| JP2010540932A (ja) | 2010-12-24 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| EP2300824B1 (en) | Detection of early stages and late stages hpv infection | |
| US8513398B2 (en) | Human papilloma virus probes for the diagnosis of cancer | |
| JP5819851B2 (ja) | Ｈｐｖ関連の癌の治療及びスクリーニングのための細胞に基づいた高処理能力ｈｐｖ免疫アッセイ | |
| CN101932711A (zh) | 晚期宫颈细胞筛查方法 | |
| US10031130B2 (en) | Device for high throughput detection of cervical disease | |
| CN106397583A (zh) | 一种hpv16型e7蛋白的poct荧光定量检测试剂盒及其应用 | |
| CN112430583B (zh) | 一种抗hpv e7的单克隆抗体及其细胞株和应用 | |
| CN112458061B (zh) | 一种抗hpv e6的单克隆抗体及其细胞株和应用 | |
| TWI497075B (zh) | 早期及晚期人類乳突病毒感染之偵測 | |
| CN108218983A (zh) | 抗hpv18型e7蛋白的单克隆抗体、检测试纸和检测试剂盒 | |
| KR100671825B1 (ko) | 인유두종바이러스에 의한 자궁경부암 진단키트 | |
| CN203870108U (zh) | 用于宫颈癌早期筛查的免疫层析的检测试纸 | |
| CN111458500A (zh) | 人呼吸道上皮细胞病原细胞学检测方法及试剂盒 | |
| CN109628580A (zh) | 一种用于宫颈病变预警的指标及其诊断试剂盒 | |
| Cochrane Gynaecological, Neuro-oncology and Orphan Cancer Group et al. | Low cost versus other screening tests to detect cervical cancer or precancer in developing countries | |
| CN119638824A (zh) | 一种可识别癌症标志物hpv e7蛋白的特异性抗体及其应用 | |
| CN112362874A (zh) | 一种宫颈癌筛查试剂盒 | |
| CN110117654A (zh) | 一种预测hpv阳性患者的宫颈病变发生情况的标志物(myc) |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C02 | Deemed withdrawal of patent application after publication (patent law 2001) | ||
| WD01 | Invention patent application deemed withdrawn after publication |
Open date: 20101229 |