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WO1991009049A1 - Oligonucleitides specifiques a l'helicobacter pylori - Google Patents

Oligonucleitides specifiques a l'helicobacter pylori Download PDF

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Publication number
WO1991009049A1
WO1991009049A1 PCT/GB1990/001979 GB9001979W WO9109049A1 WO 1991009049 A1 WO1991009049 A1 WO 1991009049A1 GB 9001979 W GB9001979 W GB 9001979W WO 9109049 A1 WO9109049 A1 WO 9109049A1
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Prior art keywords
pylori
oligonucleotide
sequence
dna
gly
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Ceased
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PCT/GB1990/001979
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English (en)
Inventor
Soad Tabaqchali
Christopher Leeson Clayton
Brendan William Wren
Harold Kleanthous
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3I Research Exploitation Ltd
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3I Research Exploitation Ltd
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    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
    • C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
    • C12Q1/68—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
    • C12Q1/6876—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes
    • C12Q1/6888—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes for detection or identification of organisms
    • C12Q1/689—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes for detection or identification of organisms for bacteria
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • C07K14/195—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from bacteria
    • C07K14/205—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from bacteria from Campylobacter (G)
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • C07K14/195—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from bacteria
    • C07K14/215—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from bacteria from Halobacteriaceae (F)

Definitions

  • This invention relates to oligonucleotides showing a substantially specific binding affinity towards genomic DNA from bacteria of the species Helicobacter pylori, until recently more usually known as Campyiobacter pylori, and which are potentially useful as H. pylori DNA probes and primers for the PCK amplification and detection of H. pylori genomic DNA and RNA.
  • H. pylori (previously C. pylori), a gram-negative spiral icroaerophilic bacteria, was first cultured from the human gastric mucosa in 1983 and has since been strongly implicated in the pathogenesis of gastritis and duodenal and peptic uiceration in man. further evidence, that C. pylori is responsible for gastritis i.s provided by studies showing that gastritis improves after patients- receive therapy directed against C. pylori infection, and the fact that both the frequency of gastritis and C. pylori infection increase with age. Patients colonised with C. pylori have been found to elicit a specific antibody response and the frequency of detection of C. pylori antibodies has also been found to increase with age.
  • the hi topathology of gastritis associated with C. pylori is well characterised.
  • the organism is closely associated with the mucosa in the gastric antrum and is located beneath the mucus layer.
  • C. pylori appear to adhere to mucus-secreting epithelial cells, and adhesion "pedestals" have been revealed by electron microscopy analysis.
  • Colonised gastric epithelium shows changes in appearance and there is partial or complete loss of microvilli.
  • Virulence determinants of C. pylori have not so far been identified, although a number of determinants possessed by this organism have been proposed as possible pathogenic factors.
  • high mobility by virtue of multiple flagella allow C. pylori to move rapidly by a corkscrew-like motion through highly viscous fluids such as the mucus layer of the gut which normally poses a barrier to bacteria en route to the gut epithelium.
  • SUBSTITUTESHEET extracellular urease allows the organism to metabolise urea and create an alkaline environment which enables its survival in an otherwise hostile environment, C. pylori normally being sensitive to low pH.
  • C. pylori urease has been found to be toxic to tissue culture cells in vitro due to the production of ammonia, and it is possible that the reported extracellular cytotoxin produced by C. pylori may contribute to the histopathological appearance of the gut mucosa associated with C. pylori colonisation.
  • patients colonised with C. pylori do produce an immune response to the organism, it appears that the organism is able to evade the host defences and cause persistent infection.
  • Campylobacter probes have previously been disclosed in WO 86/04422 and EP-A-0 232085 but not specifically for C. pylori.
  • DNA probes are disclosed derived from the chromosonal sequences of C. je uni and C. coli and which, respectively, are capable of hydbridising with at least 80% of bacteria from the species C. jejuni and C. coli, preferably at least 90%, with no ability to hybridise with bacteria not in the genus Campylobacter. This would seem to suggest some ability to hybridise with Campylobacter other than C. jejuni and C. coli, but few examples are given and there is no suggestion of any hybridisation ability at all towards C. pylori, but in any case such probes would seem somewhat lacking in species specificity and, for that reason, of little value as a diagnostic tool.
  • EP-A-0232085 discloses DNA probes capable of hybridising to the rRNA of a variety of Campylobacter species, but not including C ⁇ pylori.
  • the species mentioned are C. jejuni, C. coli, C. fetus, C. laridis, C. fetus subsp. venerialis and C. hyointestinalis.
  • the preferred probes are complementary to the RNA of the 5S, 16S oi 23S rRNA of Campylobacter, and especially to a sequence of 15 or more base pairs of the 16S rRNA sequence of C. jejuni, the base pair sequence oi which is set out in the specification.
  • C. pylori probes are not disclosed, and the probes that are disclosed would seem to lack the specificity essential to diagnostic work.
  • Campylobacter probes capable of specifically hybridizing to rRNA of C. jejuni, C. coli and C. laridis are also disclosed in EP-A-03502G5 published after the present priority date. Whilst the majority of probes disclosed in that application show a substantial measure of specificity for the sub-group of Campylobacter species consisting of the group: C. jejuni, c. coli and C. laridis, at ieast one probe is disclosed which is far less specific and, in fact, snows a wide- hybridisation ability with rRNA from other microorganisms not only of the genus Campylobacter. but also of other genera, and included within that listing is the ability to hybridise with rRNA from microorganisms of the species C.
  • probe No. 1105 is, however, hardly specific to the- selected group of C. jejuni, C. coli. and C. laridis, let alone C. pylori.
  • EP-A-0 350 392 falls in the same category as EP- ⁇ - 0 350 205 above.
  • oligonucleotide probes are disclosed having specificity towards DNA and RNA from microorganisms of the species €_. coli, C. jejuni, C. fetus. C. laridis and C. upsaliensis.
  • C. pylori probes are not disclosed.
  • SUBSTITUTE SHEET for the detection of C. pylori infection in animals, and particularly in humans, and the present invention seeks to fulfil that need, and is based on the identification and decoding of C. pylori urease gene fragment, the identification and isolation of which the present inventors reported in a paper entitled "Molecular Cloning and Expression of Campylobacter pylori Species Specific Antigens in Escherichia coli K-12" published In Infection and Immunity, 57, No. 3, 623-629 (1989). In this paper the present inventors disclosed the construction of a gene bank of C. pylori DNA in E.
  • the 2.7 kb Taql DNA fragment of C. pylori referred to from hereon in its revised classification as H. pylori, encoding the 66-kDA and 31-kDA H. pylori antigens, the A and B sub-units of H_. pylori urease, has been sequenced, giving rise to the possibility of constructing selected oligonucleotide sequences specific to H. pylori.
  • the complete sequence of the 2.7 kb Taql DNA fragment and the deduced amino acid sequence are set out in an appendix to and forming part of the present specification. The sequence corresponds substantially to the sequence disclosed in International Publication No. W0 90/04030 published after the present priority date.
  • That sequence shows two large open reading frames, codons 1 to 717 and 721 to 2400, encoding, respectively, proteins of calculated molecular weights 26.657 kDA (sub-unit A) and 60.473 (sub-unit B).
  • regions of the gene containing the most highly conserved sequences are the regions nt. nos 255-714, 720-1020 and 1950 to 2397 inclusive, and especially the regions from about nt 286 to 714 and about nt 207 ⁇ to
  • oligonucleotide probes and primers are now provided showing substantially specific binding affinity towards the ⁇ l. pylori gene encoding the A and B sub-units of the H. pylori urease gene, such oligonucleotides having a chain length of from 15 to 50- nucleotides, preferably 15-30, most preferably 15-25, and comprising a sequence of at least 15 nucleotides which is th same as or complementary to a sequence of 15 or more nucleotides selected from any one of the above mentioned numbered nt sequences.
  • oligonucleotides derived from these regions of the gene are substantially specific to genomic DNA of all strains ⁇ f H. pylori so far tested, with little or no cross-affinity to other urease positive enterobacter species. Such oligonucleotides are therefore highly preferred in the detection and identification of H. pylori infection in humans, for example, by means of saliva tests.
  • oligonucleotides having a chain length of from 15 - 30 nucleotides especially 15 - 24 and containing a sequence of at least 15 nucleotides corresponding or complementary to a sequence of at least .15 nucleotides commencing at about nt 298> or terminating at nt 714 and reproducing by PCR a fragment of 411 bp or thereabouts, or commencing at about nt. 2074 or terminating at nt. 2397 and reproducing by PCR a fragment of about 323 bp.
  • pairs of oligonucleotides of 15 to 30 nucleotides in length with one ⁇ je__ ⁇ _per of the pair having a sequence the same as or complementary to the sequence starting at about nt. 298 or 304 and the other the same as or complementary to the sequence terminating at nt. 714 have been found to amplify a 411 to 417 bp fragment that is common to all H. pyl ⁇ ri strains so far tested, that does not occur in other ureases, and can
  • SUBSTITUTE SHEET therefore be regarded as a specific marker for li. pylori. Similar results are obtainable with primer pairs amplifying a 323 bp fragment, and a characteristic 110 bp fragment.
  • the oligonucleotides of the present invention will generally have a chain length (excluding any added tail, see later) of from 15 to 50 nucleotides.
  • a chain length excluding any added tail, see later
  • Various specific sequences are given hereinafter although it is to be understood that these are given merely by way of exemplification.
  • the final selection of a particular probe, or pair of probes will depend upon a number of factors, well understood in the art, and including amongst others the stringency requirements, i.e. the ability or otherwise of the probe to tolerate local mismatching with the complementary sequence ii. the target DNA. Obviously the longer the probe the better the ability to withstand local mismatch without adversely affecting the hybridisatio of the probe to the target DNA. However, the. length of the probe always has to be balanced against other factors such as ease of synthesis. The factors affecting that choice are, however, well recognised and well within the capabilities of the person skilled in the art.
  • oligonucleotide sequences given herein as DNA sequences can equally well be constructed as RNA sequences with uracil (U) replacing thymine (T).
  • the generality of the present invention extends to DNA (and RNA) probes complementary to any sequence of nucleotide bases to be found in the highly conserved regions of the 2.7kb sequence set out hereinafter, certain sequences and pairs of sequences can be identified as being particularly preferred.
  • the pair of sequences (HPU1) 5'-GCCAATGGTA AATTAGTT-3' (nt. nos 304 to 321) and (HPU2) 5'-CTCCTTAATT GTTTTTAC-3 ' (complementary to nt. nos 697 to 714).
  • a 411 base pair product has been amplified from urease gene A (nt 304-714) using a 26 cycle PCR, which allowed visualisation of the amplified product within 5 hours.
  • Supernatants of 40 boiled H. pylori strains so far examined have given the 411bp amplified product on agarose gel electrophoresis.
  • Helicobacter mustelae and other urease positive bacteria have been found to be PCR negative.
  • HPU2 1 5' CTCCTTAATT GTTTTTACAT ACTT 3' (complementary to nt. nos 691 to 714)
  • HPU3 5' TTTGAAGTGA ATAGATGCTT AGAC 3' (nt. nos. 442 to 465)
  • HPI ⁇ 4 5' GGCTTGCCTA TCAACCAACG CGTT 3' (complementary to nt. nos 595 to 618)
  • HPU5 5' GGCCGGTTCA TCGCATTGAG TCAA 3' (nt.
  • HPU6 5' CTTTATTGGC TGGTTTAGAG TTAC 3' (complementary to nt. nos 2374 to 2397) HPUIL 5' GGGCTTGAAA GACAAGTGTT GCCG 3' (nt. nos 2242 to 2265) HPU7 5' TACAGAGAAA TGTTCGCTCA TCAT 3' (nt. nos 2143 to 2166)
  • HPU11 5' TTTTACCGGC AACACTTGTC TTTC 3' (complementary to nt. nos 2248 to 2271)
  • HPU13 5' AATGCCTTTG TCATAAGCCG CTTG 3' (complementary to nt. nos 2206 to 2229)
  • HPUV, HPU2 1 and HPUIS' are the same sequences as HPU1 , HPL2 and HPUIS above, but increased in length by six nucleotides to a total of 24).
  • oligonucleotide sequences are readily assembled using known oligonucleotide synthesis techniques.
  • Primer pair 040 + 039 has been shown to be useful on fresh gastric biopsies and amplify small fragments of 110bp and 127 bp respectively.
  • Primer pairs 018 + 054 and 017 + 055 have been shown to be 5 useful on fresh and stored paraffin embedded gastric biopsies.
  • iii The following particular H. pylori DNA sequences are amplified by the following pairs:
  • a particularly sensitive method of H. pylori detection according 15 to the invention by PCR amplification of genomic H. pylori DNA involves using two pairs of primers to amplify overlapping sequences of PL pylori DNA, the first pair of primers amplifying a first sequence of a given length, and the second pair of primers " amplifying a shorter sequence within the overall length of the first sequence, in the above 0 table certain pairings are identified by the pairs of superscripts (1,1), (2,2), (3,3) and (4,4) and these represent overlapping pairs of amplified sequences with the shorter sequence nesting within the longer sequence.
  • the 177bp sequence amplified by primers HPU3 and HPU4 nests within (i.e. is identical to an intermediate length of) the longer 417bp sequence amplified by primers HPU1 AND HPU2.
  • the 192bp product of HPU7 and HPU8 nests within the longer 324 bp product of HPU5 and HPU6; the 108bp product of HPU10 and HPUIS nests within the 234bp 0 product of HPU9 and HPU2; and the 87bp product of HPU7 and HPU13 nests within the 198bp product of HPU11 and HPU5.
  • H. pylori specific oligonucleotides of this invention may be provided with a variety of different labels: radioactive, fluorescent, enzyme, all permitting the detection of any hybridised oligonucleotide bound to the unidentified DNA sample under investigation.
  • the H. pylori specific oligonucleotides of this invention may be immobilized in any known appropriate fashion, e.g.
  • the Ii. pylori specific oligonucleotides covered by this invention include both single- and double-stranded versions, it being understood that in any subsequent hybridisation procedures such as the detection of H. pylori in the gastric mucosa or other secretions or products of the gastrointestinal tract, such double-stranded probes will require denaturing to provide the probes in single-stranded form.
  • 037 was synthesized on an Applied Biosystems oligonucleotide synthesizer and radiolabelled .at its 5' end with T4 poiynucleotoide kinase and F P and then purified.
  • Radioactively labelled probe numbers 016 to 054 were constructed in identical fashion.
  • Oligonucleotide primers e.g. 040 and 039 were used at a final concentration of 1 ⁇ M to amplify H. pylori DNA obtained by boiling H ⁇ p ___v. lori cells in 50 ul of H- _, . 0 for 10 minutes.
  • the DNA was amplified using a cycle profile of 94 ⁇ 1 min. , 31°C 1 min., and 72 C C 3 min. After the last cycie the polymerisation step was extended from 3 to 10 minutes. Twenty six cycle of amplification were performed in total. The reactions were carried out in 100 ⁇ l volumes, 20 ⁇ l samples were run on agarose gels and amplified DNA detected by ethidium bromide staining and comparison with mol.
  • H. pylori DNA has been amplified from cell populations as low as 10 H. pylori cells, . thus indicating the extreme sensitivity of H. pylori DNA probes according to this invention.
  • Simultaneous controls with C. jejuni and C. coli have failed to produce any evidence of amplification, indicating the specificity of probes according to the present invention of H. pylori.
  • pylori DNA is obtained both by hybridisation with internal oligonucleotide probes and by using two internal primers for PCR after initial amplification, e.g. by using probes 016 and 037, to give amplified fragments of 1.1 kb, and then using 5% of this reaction product to perform furtner PCR with probes 040 and 039 to give a 0.4 kb fragment.
  • Example 2 was repeated using the two primers (HPU1) 5'-GCCATGGTA AATTAGTT-3' and (HPU2) 5'-CTCCTTAATT GTTTTAC-3' amplifying the 411 bp fragment of H. pylori urease gene (nucleotide sequence nos: 304-714) on various dilutions of H. pylori, the urease clone DNA pTCP3, and 6 human gastric biopsy samples, but with a slightly modified PCR cycle, namely:
  • Lane 1 represents the urease clone DNA pTCP3; lanes 2-8
  • a similar 411b ⁇ DNA fragment has been shown to be amplified from a total of 40 different H. pylori strains. ii. mustelae and other urease positive bacteria are PCR negative under similar conditions.
  • the separated 411 bp bands were transferred by blotting and probed with a 32 D labelled oligonucleotide probe HPUIS (nt. nos 559 to 576) which is internal to the 411 bp fragment amplified by HPU1 and HPU2. after washing to remove unbound probe the bound probe was visualised by exposure to a photographic plate.
  • HPUIS oligonucleotide probe
  • PCR products were amplified from paraffin stored gastric biopsy sections (section nos. 4593, 10412, 7213, 4591 and 1841 - Lanes 3 - 7 Figure 4).
  • Lane 2 represents a positive control paraffin embedded H. pylori 630, Lane 9 a negative control and Lane 8 a 1kb mol. wt. standard ladder.
  • Lane 1 shows a PCR product of 100 bp amplified from section 4593 by human ⁇ -globulin primers.
  • Lanes 3 - 6 clearly show the 110 bp fragment amplified by the primers HPU18 and HPU54 confirming the presence of H. pylori in sections 4594 10412 7213 4591 (Lanes 3 - 6) and its absence from section 1841 (Lane 7).
  • Sequence Type Nucleotide with corresponding amino acid Sequence Length: 2767 bp Strandedness: Double Topology: Linear Molecule Type: Genomic DNA Original Source Organism: H. pylori Immediate Experimental Source:
  • H. pylori urease gene A and B sub-units Properties: H. pylori urease gene A and B sub-units.

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Abstract

On décrit des séquences d'oligonucléotides spécifiques à l'uréase de l'helicobacter pylori et utiles comme sondes et initiateurs ADN pour la détection de l'infection par l'helicobacter pylori chex les humains.
PCT/GB1990/001979 1989-12-19 1990-12-19 Oligonucleitides specifiques a l'helicobacter pylori Ceased WO1991009049A1 (fr)

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GB898928625A GB8928625D0 (en) 1989-12-19 1989-12-19 H.pylori dna probes
GB8928625.6 1989-12-19

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WO1991009049A1 true WO1991009049A1 (fr) 1991-06-27

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Cited By (17)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO1993007273A1 (fr) * 1991-10-03 1993-04-15 Institut Pasteur Genes d'helicobacter pylori necessaires pour la regulation et la maturation de l'urease et leur utilisation
WO1993018150A1 (fr) * 1992-03-02 1993-09-16 Biocine S.P.A. Proteines d'helicobacter pylori utiles pour des vaccins et des diagnostics
WO1994026901A1 (fr) * 1993-05-19 1994-11-24 Institut Pasteur Compositions immunogenes destinees a proteger contre les infections a helicobacter, polypeptides utilises dans lesdites compositions et sequences d'acides nucleiques codant lesdits polypeptides
WO1996001273A1 (fr) * 1994-07-01 1996-01-18 Rican Limited Preparation a base de proteine antigene d'helicobacter pylori et dosage immunologique
WO1998021225A1 (fr) * 1996-11-14 1998-05-22 Merieux Oravax Polypeptides helicobacter et molecules de polynucleotides correspondantes
US5837502A (en) * 1993-06-30 1998-11-17 Reckitt & Colman Products Limited Helicobacter pylori haemagglutinin protease protein, nucleic acid encoding therefor and antibodies specific thereto
US5843460A (en) * 1993-05-19 1998-12-01 Institut Pasteur Immunogenic compositions against helicobacter infection, polypeptides for use in the compositions, and nucleic acid sequences encoding said polypeptides
US6090611A (en) * 1992-03-02 2000-07-18 Chiron S.P.A. Helicobacter pylori proteins useful for vaccines and diagnostics
US6130059A (en) * 1992-03-02 2000-10-10 Covacci; Antonello Helicobacter pylori proteins useful for vaccines and diagnostics
US6190667B1 (en) 1998-06-30 2001-02-20 Institut Pasteur Methods of inhibiting Helicobacter pylori
EP0977482A4 (fr) * 1997-04-01 2002-03-06 Merieux Oravax Identification de polynucleotides codant de nouveaux polypeptides helicobacter dans le genome helicobacter
EP0965646A4 (fr) * 1996-07-24 2003-01-02 Fuso Pharmaceutical Ind Sondes permettant de detecter et d'identifier helicobacter pylori
US6911308B2 (en) * 2001-01-05 2005-06-28 Exact Sciences Corporation Methods for detecting, grading or monitoring an H. pylori infection
US7141244B1 (en) 1992-03-02 2006-11-28 Chiron Srl Helicobacter pylori proteins useful for vaccines and diagnostics
US7901907B2 (en) 1996-01-04 2011-03-08 The Provost Fellows And Scholars Of The College Of The Holy And Undivided Trinity Of Queen Elizabeth Near Dublin Process for production of Helicobacter pylori bacterioferritin
CN111944916A (zh) * 2020-09-14 2020-11-17 壹宏(深圳)基因有限公司 用于粪便微生物检测的试剂及其在胃幽门螺旋杆菌检测的应用
CN113943822A (zh) * 2021-09-27 2022-01-18 山东第一医科大学(山东省医学科学院) 一种基于lamp-lfd的幽门螺杆菌poct检测试剂盒及方法

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Publication number Priority date Publication date Assignee Title
EP0367644A1 (fr) * 1988-10-06 1990-05-09 Institut Pasteur Séquence de nucléotides codant pour une proteine à activité uréasique

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EP0367644A1 (fr) * 1988-10-06 1990-05-09 Institut Pasteur Séquence de nucléotides codant pour une proteine à activité uréasique

Non-Patent Citations (2)

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Title
Chemical Abstracts, vol. 109, no. 7, August 1988, (Columbus, Ohio, US), H.L.T. Mobley et al.: "Characterization of urease from Campylobacter pylori", see page 289 *
Infection and Immunity, vol. 57, no. 2, Febraury 1989, American Society for Microbology, A.L. Clayton et al.: "Molecular cloning and expression of Campylobacter pylori species-specific antigens in Escherichia coli K-12", pages 623-629 *

Cited By (34)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO1993007273A1 (fr) * 1991-10-03 1993-04-15 Institut Pasteur Genes d'helicobacter pylori necessaires pour la regulation et la maturation de l'urease et leur utilisation
US6271017B1 (en) 1991-10-03 2001-08-07 Institut Pasteur Genes of Heliciobacter pylori necessary for the regulation and maturation of urease and their use
US6027878A (en) * 1991-10-03 2000-02-22 Institut Pasteur Genes of Helicobacter pylori necessary for the regulation and maturation of urease and their use
US5986051A (en) * 1991-10-03 1999-11-16 Institut Pasteur Genes of Helicobacter pylori necessary for the regulation and maturation of urease and their use
WO1993018150A1 (fr) * 1992-03-02 1993-09-16 Biocine S.P.A. Proteines d'helicobacter pylori utiles pour des vaccins et des diagnostics
US7141244B1 (en) 1992-03-02 2006-11-28 Chiron Srl Helicobacter pylori proteins useful for vaccines and diagnostics
US7700755B2 (en) 1992-03-02 2010-04-20 Novartis Vaccines And Diagnostics S. R. L. Helicobacter pylori CAI antigen
US6130059A (en) * 1992-03-02 2000-10-10 Covacci; Antonello Helicobacter pylori proteins useful for vaccines and diagnostics
US6090611A (en) * 1992-03-02 2000-07-18 Chiron S.P.A. Helicobacter pylori proteins useful for vaccines and diagnostics
US6077706A (en) * 1992-03-02 2000-06-20 Chiron Corporation Helicobacter pylori proteins useful for vaccines and diagnostics
US5843460A (en) * 1993-05-19 1998-12-01 Institut Pasteur Immunogenic compositions against helicobacter infection, polypeptides for use in the compositions, and nucleic acid sequences encoding said polypeptides
US6248330B1 (en) 1993-05-19 2001-06-19 Institut Pasteur Immunogenic compositions against helicobacter infection, polypeptides for use in the compositions, and nucleic acid sequences encoding said polypeptides
US6258359B1 (en) 1993-05-19 2001-07-10 Institut Pasteur Immunogenic compositions against helicobacter infection, polypeptides for use in the compositions, and nucleic acid sequences encoding said polypeptides
WO1995014093A1 (fr) * 1993-05-19 1995-05-26 Institut Pasteur Compositions immunogenes dirigees contre les infections par helicobacter, polypeptides utilisables dans ces compositions, et sequences d'acides nucleiques codant ces polypeptides
WO1994026901A1 (fr) * 1993-05-19 1994-11-24 Institut Pasteur Compositions immunogenes destinees a proteger contre les infections a helicobacter, polypeptides utilises dans lesdites compositions et sequences d'acides nucleiques codant lesdits polypeptides
US5837502A (en) * 1993-06-30 1998-11-17 Reckitt & Colman Products Limited Helicobacter pylori haemagglutinin protease protein, nucleic acid encoding therefor and antibodies specific thereto
GB2303855B (en) * 1994-07-01 1998-10-28 Rican Limited Helicobacter pylori antigenic protein preparation and immunoassays
AU696941B2 (en) * 1994-07-01 1998-09-24 Rican Limited Helicobacter pylori antigenic protein preparation and immunoassays
GB2303855A (en) * 1994-07-01 1997-03-05 Rican Limited Helicobacter pylori antigenic protein preparation and immunoassays
WO1996001273A1 (fr) * 1994-07-01 1996-01-18 Rican Limited Preparation a base de proteine antigene d'helicobacter pylori et dosage immunologique
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CN113943822A (zh) * 2021-09-27 2022-01-18 山东第一医科大学(山东省医学科学院) 一种基于lamp-lfd的幽门螺杆菌poct检测试剂盒及方法
CN113943822B (zh) * 2021-09-27 2024-03-12 山东第一医科大学(山东省医学科学院) 一种基于lamp-lfd的幽门螺杆菌poct检测试剂盒及方法

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