CN114264816A - Echinococcus antibody immunochromatographic test strip and detection card - Google Patents
Echinococcus antibody immunochromatographic test strip and detection card Download PDFInfo
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- CN114264816A CN114264816A CN202111601648.0A CN202111601648A CN114264816A CN 114264816 A CN114264816 A CN 114264816A CN 202111601648 A CN202111601648 A CN 202111601648A CN 114264816 A CN114264816 A CN 114264816A
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- 238000001514 detection method Methods 0.000 title claims abstract description 149
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Abstract
本发明公开了一种棘球蚴抗体免疫层析试纸条和检测卡。该试纸条包括硝酸纤维素膜；硝酸纤维素膜上设置有检测线组和质控线；检测线组包括分别包被不同重组棘球蚴抗原的T1检测线、T2检测线和T3检测线；结果判定规则为：①质控线和任意2条或以上检测线都显色，判定结果为阳性；②质控线显色而检测线均不显色，或者质控线显色而检测线仅一条显色，判定结果为阴性；③质控线不显色，无论检测线是否显色，均表示所述棘球蚴抗体免疫层析试纸条失效，应该重新检测。该检测卡包括壳体和所述试纸条；壳体上依次设置有：登记记录区、观察窗口、加样孔。本发明对包虫病的检测准确度高，结构简单合理，便于独立包装和长期保存。
The invention discloses an echinococcosis antibody immunochromatographic test strip and a detection card. The test strip includes a nitrocellulose membrane; a detection line group and a quality control line are arranged on the nitrocellulose membrane; the detection line group includes T1 detection line, T2 detection line and T3 detection line respectively coated with different recombinant echinococcosis antigens ; The result judgment rule is: ① The quality control line and any two or more test lines develop color, and the judgment result is positive; ② The quality control line develops color but none of the test lines develops color, or the quality control line develops color and the test line develops color. If only one color develops, the judgment result is negative; ③ The quality control line does not develop color, no matter whether the detection line develops color or not, it means that the echinococcosis antibody immunochromatographic test strip is invalid and should be re-tested. The detection card includes a casing and the test strip; the casing is sequentially provided with: a registration and recording area, an observation window, and a sample adding hole. The invention has high detection accuracy for hydatid disease, simple and reasonable structure, and is convenient for independent packaging and long-term storage.
Description
Technical Field
The invention relates to a medical detection device, in particular to an echinococcus antibody immunochromatographic test strip and a detection card.
Background
Echinococcosis (also called echinococcosis) is a parasitic disease which is caused by the parasitosis of echinococcus and its larva (echinococcosis) in human and animal bodies and is a kind of parasitosis which is harmful to human and animal. Among the echinococcosis diseases known so far, the most serious of them to human and socioeconomic are echinococcosis granulosa (CE) and echinococcosis multocida (AE). The disease is highly epidemic in Qinghai-Tibet plateau areas in China, and is a parasitic disease which is mainly prevented and treated in China. The problems of underdeveloped economy, severe natural conditions, inconvenient traffic, high working cost and shortage of health professionals exist in the epidemic area generally. The technical level and the working efficiency of prevention and treatment need to be improved.
Echinococcus is parasitic in human body in a closed manner, grows like a tumor, and mainly infects liver (accounting for about 70% of clinical cases). The currently preferred diagnostic method is medical imaging, and is most widely used in B-mode ultrasound. However, the method has high requirements on the technical level and experience of inspectors, and has high clinical misdiagnosis rate for atypical cysts smaller than 2cm, and an immunological method is required for auxiliary diagnosis. The sensitivity and specificity of immunological detection also play an important role in the timely discovery of patients and follow-up after treatment; the kit has more important reference values for epidemiological investigation of the epidemiological area, formulation of prevention and treatment planning and assessment and immunological diagnosis of prevention and treatment effects. The current echinococcus antibody detection reagent is most commonly used and sensitive by an ELISA method, but the method needs 2-3 hours to obtain a detection result, is long in use time, needs certain experimental equipment and experimental technicians to operate, and is difficult to use for on-site investigation of echinococcosis in remote areas. The immunochromatography rapid-detection test strip method is more practical than the ELISA method, has better sensitivity and specificity, is simpler, more convenient and quicker to operate, does not need special instruments and equipment, is easy to interpret results, and is particularly suitable for basic clinical detection and epidemiological investigation.
At present, most of echinococcus antibody diagnostic reagents sold in the market use natural parasite antigens, the natural materials are not convenient to obtain and difficult to standardize, so that the reagent has large batch-to-batch difference, has high cross reaction with other parasitic diseases such as cysticercosis and the like, and has poor diagnosis specificity and stability. In some researches, recombinant antigens are used to replace natural antigens, but the detection sensitivity is low, because only a single antigen is used for detection in the detection, the detection sensitivity is poor, and the detection is easy to miss. Therefore, the recombinant antigens leading to these reports could not be practically applied to the detection of echinococcosis; or must be combined with the native antigen for detection.
Disclosure of Invention
In order to improve the detection accuracy of echinococcosis, the invention discloses an echinococcosis antibody immunochromatographic test strip, which comprises a nitrocellulose membrane; the nitrocellulose membrane is provided with a detection line group and a quality control line; the detection line group comprises a T1 detection line, a T2 detection line and a T3 detection line which are respectively coated with different recombinant echinococcus antigens; the result determination rule is: the quality control line and any 2 or more detection lines are colored, and the result is judged to be positive; secondly, the quality control line is colored, but the detection line is not colored, or the quality control line is colored, but only one detection line is colored, and the judgment result is negative; and thirdly, the quality control line does not develop color, whether the detection line develops color or not indicates that the echinococcus antibody immunochromatographic test strip fails, and the echinococcus antibody immunochromatographic test strip should be re-detected.
In some embodiments, a gold-labeled pad is further included; the gold-labeled pad is a glass fiber membrane, and is fixed with colloidal gold-labeled streptococcus G protein or colloidal gold-labeled staphylococcus aureus protein A; when the gold label pad is fixed with the colloidal gold labeled streptococcus G protein, the quality control line is coated with staphylococcus aureus protein A; when the gold label pad is fixed with the colloidal gold labeled staphylococcus aureus protein A, the quality control line is coated with streptococcus G protein; the gold label pad is connected with the end part of the nitrocellulose membrane in an overlapping way; the recombinant Echinococcus antigen includes recombinant Echinococcus antigen Em18 (reference: Jiang L, Xu XN, Li X, Xue HC, Feng Z.identification of the immunogenic regions of the Em18antigen and expressed mammalian specificity for extracellular infection [ J ]. Parasite antigen, 2004, 26 (10): 377-85), recombinant Echinococcus antigen Egcystatin (an Echinococcus granulosus cysteine protease inhibitor (issued patent: a novel Echinococcus diagnostic antigen and use thereof; 2016-2-4, ZL, 201610081438.6) and recombinant Echinococcus antigen B1 (reference: Agents MB, Rornus RNA, amino group V, Farne J.S.origin, amino group J.induction, amino group antigen, protein of Escherichia coli, Escherichia coli antigen, J.2000, amino group antigen of Escherichia coli antigen, Escherichia coli antigen, Escherichia coli antigen, Escherichia coli antigen, Escherichia coli, Escherichia coli, Escherichia, 75(3): 331-40).
In some embodiments, further comprising a sample pad, absorbent pad, and a base plate; the sample pad, the gold label pad, the nitrocellulose membrane and the absorbent pad are sequentially connected to the bottom plate in an end overlapping manner, wherein two ends of the nitrocellulose membrane are arranged at positions close to the bottom plate in the overlapping manner; the distance from the detection line group to the gold mark pad is shorter than the distance from the quality control line to the gold mark pad; the sample pad is a blood filtering membrane; the sample to be tested is selected from one or more of serum, plasma, whole blood, tissue fluid, urine, saliva and feces.
On the other hand, the invention also discloses an echinococcus antibody immunochromatographic test card, which comprises a shell and the echinococcus antibody immunochromatographic test strip; the casing is provided with:
a registration recording area for recording information including sample information, a detection date, and a detection result;
the observation window is used for observing the immunochromatography condition of the test strip arranged in the shell;
and the sample adding hole is used for adding a sample to the test strip arranged in the shell.
Further, a hollow hole is formed in the registration recording area.
In some embodiments, the housing is further provided with a raised spot region for holding the immunochromatographic detection device; the housing includes a first end and a second end; the projection point area, the registration recording area, the observation window, and the sample application hole are sequentially provided between the first end portion and the second end portion.
In some embodiments, the bottom of the inner surface of the housing is provided with a test strip positioning plate and a test strip side positioning column; the test strip side positioning columns are arranged on two sides of the test strip positioning plate; the top of the inner surface of the shell is provided with a test strip top positioning column, so that the test strip top positioning column can be pressed on the test strip positioning plate.
In some embodiments, the number of the test strip side positioning columns is 4-10, and the test strip side positioning columns are equally divided into 2 rows and arranged on two sides of the test strip positioning plate.
Preferably, the number of the test strip side positioning columns is 6, and the test strip side positioning columns are equally divided into 2 rows and arranged on two sides of the test strip positioning plate.
Further, the housing includes: a closed configuration for detection and transport; the opening state is used for taking and placing the test strip.
In some embodiments, the housing comprises an upper housing and a lower housing; a clamping shell positioning column is arranged on the inner surface of the upper shell; the inner surface of the lower shell is provided with a clamping shell positioning hole in a matching mode, the clamping shell positioning hole is used for being inserted into the clamping shell positioning column and connecting the upper shell and the lower shell, and therefore the shells are in a closed state. When the positioning column of the card shell is pulled out from the positioning hole of the card shell, the shell is in an opening shape.
In one or more embodiments, the housing comprises an upper housing and a lower housing; one side of the upper shell is connected with one side of the lower shell through a hinge, and the upper shell covers the lower shell, so that the shell is in a closed state.
In some embodiments, the number of the card shell positioning holes and the number of the card shell positioning columns are the same and are 4-12. Preferably, the number of the clamping shell positioning holes and the number of the clamping shell positioning columns are the same and are 8.
In some embodiments, the sample application hole is in a funnel shape with a wide top and a narrow bottom, and is opposite to a sample pad of the test strip arranged in the shell.
In some embodiments, the detection line and the quality control line are observable through the observation window. The sample information includes one or more of a sample name, a sample source, and a sample number. A transparent plate is arranged on the detection window.
Further, the hollow holes are arranged in two rows, and the registration recording area is divided into three rows which are respectively used for recording the sample information, the detection date and the detection result.
Further, the convex point area is positioned at the upper left corner of the upper surface of the shell and is shaped like an 1/4 circular arc.
The invention adopts 3 recombinant echinococcus antigens to prepare the test strip, thereby stabilizing the quality of the diagnostic reagent. The human serum of the cysticercosis is detected by respectively adopting a commercial ELISA echinococcosis antibody detection kit and the echinococcosis antibody immunochromatographic detection card of the invention, the cross reaction of the ELISA kit and the cysticercosis is 67 percent (18/27), but no cross reaction is detected by the invention (0/20), so the cross reaction with other parasites such as cysticercosis can be reduced by adopting the recombinant echinococcosis antigen, and the diagnostic efficiency and the standardization of the reagent are improved. By the combined detection of the three antigens and the adoption of a new detection result judgment rule, the detection accuracy is improved. Meanwhile, the detection time is only less than 30 minutes, and whole blood, serum and plasma samples can be detected, so that the diagnosis efficiency is greatly improved, and the detection operation is facilitated.
In the present invention, 3 detection antigens (i.e., detection lines) are used for detection, and it is specified that any 2 or more detection lines among them are colored, and the detection is determined to be positive. Compared with single recombinant antigen detection, the determination method improves the accuracy of the detection result, and the sensitivity and the specificity of the detection result have statistical difference (chi)Sensitivity of the composition 2＝10.714，P<0.05；χSpecificity of 2＝4.138，P<0.05). The invention has the advantages of simple and rapid detection operation, no need of special instruments and equipment, and easy result interpretation.
The invention provides an echinococcus antibody immunochromatographic assay detection card, wherein a single test strip adopts an independent packaging specification, which is beneficial to the independent packaging and long-term storage of the single test strip for detecting echinococcosis antibody; by adopting various recombinant antigens to replace natural antigens for detecting the echinococcosis antibody, the antigen is simple and convenient to produce and prepare, easy to control the quality, low in production cost and beneficial to improving the diagnostic efficiency and standardization of the reagent; simultaneously, the sample information and the detection result are recorded, so that the image-text information is conveniently filed and searched; the arrangement of the observation window is favorable for observing the detection result; the funnel-shaped sample adding hole is beneficial to the concentrated collection of the sample and the buffer solution on the sample pad without side leakage; the upper shell and the lower shell are matched, so that the test strip can be packed independently and stored for a long time; the test strip positioning plate, the test strip side positioning column and the test strip top positioning column are beneficial to fixing the test strip, and the test strip is prevented from shaking up and down, left and right and deviating in the transportation process; the hollow hole can communicate the test strip water absorption pad with the outside air, is favorable for volatilizing with the synchronous moisture of nitrocellulose membrane, and avoids the liquid on the water absorption pad from flowing back to the nitrocellulose membrane to cause the background color of a test strip detection area (the area where the detection line is located) to turn red.
The echinococcus antibody immunochromatographic test strip and the detection card are used for detecting echinococcosis patient serum antibody by using the kit, and a plurality of recombinant antigens are coated to detect echinococcosis antibody. The test strip has a simple and reasonable structure, is easy to produce in batches, package independently and store for a long time, is suitable for on-site rapid diagnosis of the echinococcosis in an epidemic area, has high detection accuracy and low detection cost, and is beneficial to screening the echinococcosis antibodies of large-scale crowds. When the invention is used, a plurality of recombinant antigens are adopted to replace natural antigens, the quality of the diagnostic reagent is stabilized, the cross reaction with other parasitic diseases such as cysticercosis and the like is reduced, and the diagnostic efficiency and the standardization of the reagent are improved. Meanwhile, the detection time is within 30 minutes, and the diagnosis efficiency is greatly improved.
The invention receives the subsidy of the following fund: the national research and development project (No.2021YFC2300800,2021YFC2300803), the national health Committee prevention and treatment research focus laboratory (No.2021WZK1003), and the Shanghai Weijian project (No.201840133, No. 201940302).
The conception, the specific structure and the technical effects of the present invention will be further described with reference to the accompanying drawings to fully understand the objects, the features and the effects of the present invention.
Drawings
FIG. 1 is the top view and the side view of an echinococcus antibody immunochromatographic test strip.
FIG. 2 is a schematic diagram of an echinococcus antibody immunochromatographic test strip.
FIG. 3 is the schematic diagram of the whole internal structure of the echinococcus antibody immunochromatography detection card.
FIG. 4 is the assembling diagram of the echinococcus antibody immunochromatography detection card.
FIG. 5 is the schematic diagram of the upper and lower shells of the echinococcus antibody immunochromatography detection card.
FIG. 6 is a perspective view of an echinococcus antibody immunochromatographic detection card.
Wherein, 1-test paper strip; 2-a shell; 3-nitrocellulose membrane; 4-sample pad; 5-water absorption pad; 6-a bottom plate; 7-gold label pad; 8-detection line group; 9-quality control line; 10-a lower shell; 11-an upper shell; 12-a viewing window; 13-a sample application well; 14-card housing locating posts; 15-clamping shell positioning holes; 16-test strip side positioning column; 17-hollowed out holes; 18-test strip positioning plate; 19-registration of the recording area; 20-a salient point region; 21-a first end portion; 22-a second end; 23-test paper strip top reference column.
Detailed Description
In order to make the technical means, the characteristics, the purposes and the functions of the invention easy to understand, the invention is further described with reference to the specific drawings. However, the present invention is not limited to the following embodiments.
It should be understood that the structures, ratios, sizes, and the like shown in the drawings and described in the specification are only used for matching with the disclosure of the specification, so as to be understood and read by those skilled in the art, and are not used to limit the conditions under which the present invention can be implemented, so that the present invention has no technical significance, and any structural modification, ratio relationship change, or size adjustment should still fall within the scope of the present invention without affecting the efficacy and the achievable purpose of the present invention.
Example 1 Echinococcus antibody immunochromatographic test strip
Fig. 1 and 2 show a specific embodiment of the echinococcus antibody immunochromatographic test strip of the present invention, and in this embodiment, the echinococcus antibody immunochromatographic test strip comprises a bottom plate 6, and a sample pad 4, a gold-labeled pad 7, a nitrocellulose membrane 3, and an absorbent pad 5, which are sequentially connected to the bottom plate 6 in an end-to-end overlapping manner. Wherein both ends of the nitrocellulose membrane 3 are pressed against the bottom plate 6 at the positions which are arranged in the overlapping connection, namely one end of the gold label pad 7 and one end of the absorbent pad 5 are respectively pressed against both ends of the nitrocellulose membrane 3 (as shown in fig. 1).
The gold-labeled pad 7 is a glass fiber membrane on which colloidal gold-labeled streptococcal protein G or colloidal gold-labeled staphylococcus aureus protein A is immobilized.
The nitrocellulose membrane 3 is provided with a detection line group 8 and a quality control line 9. The distance from the detection line group 8 to the gold pad 7 is shorter than the distance from the quality control line 9 to the gold pad 7.
The detection line group 8 comprises a T1 detection line, a T2 detection line and a T3 detection line which are respectively coated with different recombinant echinococcus antigens. The recombinant echinococcus antigen comprises recombinant echinococcus antigen Em18, recombinant echinococcus antigen EgCystatin and recombinant echinococcus antigen AgB 1. The recombinant echinococcus antigen Em18, the recombinant echinococcus antigen EgCystatin and the recombinant echinococcus antigen AgB1 are matched and correspond to the T1 detection line, the T2 detection line and the T3 detection line optionally. Namely, there can be 6 combinations: 1) the T1 detection line, the T2 detection line and the T3 detection line are respectively coated with recombinant echinococcus antigen Em18, recombinant echinococcus antigen EgCystatin and recombinant echinococcus antigen AgB 1;
2) the T1 detection line, the T2 detection line and the T3 detection line are respectively coated with recombinant echinococcus antigen AgB1, recombinant echinococcus antigen Em18 and recombinant echinococcus antigen EgCystatin;
3) the T1 detection line, the T2 detection line and the T3 detection line are respectively coated with recombinant echinococcus antigen EgCystatin, recombinant echinococcus antigen AgB1 and recombinant echinococcus antigen Em 18;
4) the T1 detection line, the T2 detection line and the T3 detection line are respectively coated with recombinant echinococcus antigen EgCystatin, recombinant echinococcus antigen Em18 and recombinant echinococcus antigen AgB 1;
5) the T1 detection line, the T2 detection line and the T3 detection line are respectively coated with recombinant echinococcus antigen Em18, recombinant echinococcus antigen AgB1 and recombinant echinococcus antigen EgCystatin;
6) the T1 detection line, the T2 detection line and the T3 detection line are respectively coated with recombinant echinococcus antigen AgB1, recombinant echinococcus antigen EgCystatin and recombinant echinococcus antigen Em 18;
when the colloidal gold labeled streptococcus G protein is fixed on the gold labeled pad 7, the quality control line 9 is coated with staphylococcus aureus protein A; when the gold-labeled pad 7 is fixed with the colloidal gold-labeled staphylococcus aureus protein A, the quality control line 9 is coated with streptococcus G protein.
The sample pad 4 is a blood filtration membrane. The samples tested may be serum, plasma and whole blood.
Example 2 Echinococcus antibody immunochromatography detection card
FIGS. 3-6 show an embodiment of the invention, in which the echinococcus antibody immunochromatographic test card comprises a housing 2 and a test strip 1. The housing 2 comprises a first end 21 and a second end 22. Between the first end 21 and the second end 22, the housing 2 is provided with: a raised spot region 20 for holding the immunochromatographic detection device; a registration recording area 19 for recording information including sample information, a detection date, and a detection result; an observation window 12 for observing the immunochromatographic condition of the test strip 1 disposed in the housing 2; and a sample application hole 13 for applying a sample to the test strip 1 disposed in the housing 2.
The projection point region 20 is located at the upper left corner of the upper surface of the housing 2 and has an arc shape of 1/4.
The registration recording area 19 is provided with a hollowed hole 17. The hollow holes 17 are arranged in two rows, and divide the registration recording area 19 into three rows for recording sample information, detection date, and detection result, respectively.
The sample adding hole 13 is in a funnel shape with a wide upper part and a narrow lower part and is opposite to the sample pad 4 of the test strip 1 arranged in the shell 2.
A test strip positioning plate 18 and a test strip side positioning column 16 are arranged at the bottom of the inner surface of the shell 2; the test strip side positioning columns 16 are arranged on two sides of the test strip positioning plate 18; the top of the inner surface of the housing 2 is provided with a test strip top positioning column 23, so that the test strip top positioning column 23 can press the test strip 1 on the test strip positioning plate 18. The number of the test strip side positioning columns 16 is 6, and the test strip side positioning columns are equally divided into 2 rows and arranged on two sides of the test strip positioning plate 18.
The housing 2 includes: a closed configuration (as shown in fig. 3) for detection and transport; in the open configuration (as shown in fig. 1 and 2) for handling the test strip. The housing 2 includes an upper housing 11 and a lower housing 10; a clamping shell positioning column 14 is arranged on the inner surface of the upper shell 11; the inner surface of the lower shell 10 is provided with a matching card shell positioning hole 15 for inserting the card shell positioning column 14, and the upper shell 11 and the lower shell 10 are connected, so that the shell 2 is in a closed state. When the housing positioning posts 14 are pulled out of the housing positioning holes 15, the housing 2 is opened. The number of the clamping shell positioning holes 15 is the same as that of the clamping shell positioning columns 14, and the number of the clamping shell positioning holes is 8.
The test strip 1 is the echinococcus antibody immunochromatographic test strip in example 1.
Example 3
In this embodiment, the number of the test strip side positioning columns is 4, and the test strip side positioning columns are divided into 2 rows and arranged on two sides of the test strip positioning plate. The number of the clamping shell positioning holes and the number of the clamping shell positioning columns are the same and are 4. Otherwise, the procedure was as in example 2.
Example 4
In this embodiment, the number of the test strip side positioning columns is 10, and the test strip side positioning columns are divided into 2 rows and arranged on two sides of the test strip positioning plate. The number of the clamping shell positioning holes and the number of the clamping shell positioning columns are the same and are 12. Otherwise, the procedure was as in example 2.
Example 5
In this embodiment, the housing includes an upper housing and a lower housing; one side of the upper shell is connected with one side of the lower shell through a hinge, and the upper shell covers the lower shell, so that the shell is in a closed state. Otherwise, the procedure was as in example 2.
Preparing an echinococcus antibody immunochromatography detection card:
as described in example 1, the test strip was composed of a base plate, a sample pad, a gold-labeled pad, a nitrocellulose membrane, and a water absorbent pad. Adhering the treated sample pad, water absorption pad, nitrocellulose membrane and gold-labeled binding pad on a bottom plate (PVC rubber plate), fixing 3 recombinant antigens and staphylococcus aureus Protein A (Protein A, SPA) in a detection line (T1-T3 line) and a quality control line (C line) zone of the nitrocellulose membrane respectively by using a membrane spraying instrument, cutting the test strips into test strips with the width of 4mm by using a test strip cutting machine, putting the test strips into a prefabricated shell, putting the assembled test cards into a sealing bag containing a drying agent, and storing at room temperature for later use after sealing, wherein the effective period is as long as 2 years.
During detection, the echinococcus antibody immunochromatographic detection card is unpacked independently, and after a sample (such as serum, plasma or whole blood) is dripped into the sample adding hole 13, the sample enters the sample pad 4, and moves towards the water absorption pad 5 along the test strip 1 due to the chromatography effect. When the sample moves to the gold-labeled pad 7, the antibody in the sample binds to the colloidal gold on the gold-labeled pad 7. The combined composite continues to move towards the absorbent pad 5. When the area of the detection line group 8 is detected, the corresponding antibody in the sample is specifically combined with the corresponding recombinant antigen of the area, so that the area shows red color, and the specific immunodiagnosis is realized. The detection time is within 30 minutes, even about 15 minutes.
Before the sample is dripped, sample information such as sample source, serial number and the like can be recorded in the registration recording area 19; the sample and the buffer solution are dripped on the sample pad 4 of the test strip 1 through the sample adding hole 13; the liquid with the sample flows to the gold label pad 7 to react with the colloidal gold, then flows to the nitrocellulose membrane 3 to react with the detection line 8 and the quality control line 9 in sequence; the reaction changes of the detection line 8 and the quality control line 9 are observed and recorded by the detection personnel through the observation window 12.
Specifically, the detector can hold the convex point area 20 to move the immunochromatography detection device for detecting the echinococcosis antibody, observe the color change on the detection line group 8 and the quality control line 9 through the observation window 12 of the upper shell 11, so as to determine whether the detection sample is in positive reaction, record the detection result in the registration recording area 19, shoot the image-text information and save the image-text information.
The decision rule is as follows: positive reaction: the quality control line and any 2 or more detection lines are colored. Negative reaction: the quality control line is colored and the detection line is not colored, or the quality control line is colored and only one detection line is colored. And thirdly, the quality control line does not develop color, and whether the detection line develops color or not, the failure of the test strip is indicated, and the test strip should be detected again.
Test example 1 detection of sensitivity and specificity
The recombinant echinococcus antigen involved in the invention is not limited to Em18, EgCystatin and AgB1, but also can be other recombinant echinococcus antigens. The present invention will be described herein by taking these 3 recombinant echinococcus antigens as examples, so as to understand the beneficial effects of the present invention.
To evaluate the screening method of the present invention, 60 serum samples were used for the assay, and the results are shown in Table 1. The total amount of the collected human serum is 60 parts, wherein 30 parts of the serum of echinococcus patients diagnosed by B ultrasonic diagnosis and 30 parts of the serum of healthy human are obtained from Cuminum vulgare of Sichuan province. T1, T2 and T3 represent 3 recombinant antigens (Em 18, EgCystatin and AgB1 in this test example), respectively, and the detection sensitivity (76.67%, 33.33%, 86.67%) and the specificity (90.00%, 100.00%, 83.33%) are respectively. The first screening method is a method in which a positive reaction is judged by coloring any 1T line, and the sensitivity, specificity and accuracy index of the judgment method are (93.33%, 80.00%, 0.73), respectively. The second screening method (i.e., the screening method of the present invention) was determined to be positive by coloring at least 2 detection lines (T lines), and the sensitivity, specificity and accuracy index of the determination method were (83.33%, 96.67%, 0.80), respectively. Statistical differences in sensitivity and specificity (χ) between the two screening methods were determined by chi-square comparisonSensitivity of the composition 2＝10.714，P<0.05；χSpecificity of 2＝4.138，P<0.05), which indicates that the second screening method is more effective.
TABLE 1 comparison of different detection lines and determination methods
Note: indicates the first screening method; indicates the second screening method.
The foregoing detailed description of the preferred embodiments of the invention has been presented. It should be understood that numerous modifications and variations could be devised by those skilled in the art in light of the present teachings without departing from the inventive concepts. Therefore, the technical solutions available to those skilled in the art through logic analysis, reasoning and limited experiments based on the prior art according to the concept of the present invention should be within the scope of protection defined by the claims.
Claims (10)
1. An echinococcus antibody immunochromatographic test strip is characterized by comprising a nitrocellulose membrane; the nitrocellulose membrane is provided with a detection line group and a quality control line; the detection line group comprises a T1 detection line, a T2 detection line and a T3 detection line which are respectively coated with different recombinant echinococcus antigens; the result determination rule is: the quality control line and any 2 or more detection lines are colored, and the result is judged to be positive; secondly, the quality control line is colored, but the detection line is not colored, or the quality control line is colored, but only one detection line is colored, and the judgment result is negative; and thirdly, the quality control line does not develop color, whether the detection line develops color or not indicates that the echinococcus antibody immunochromatographic test strip fails, and the echinococcus antibody immunochromatographic test strip should be re-detected.
2. The echinococcus antibody immunochromatographic test strip of claim 1, further comprising a gold-labeled pad; the gold-labeled pad is a glass fiber membrane, and is fixed with colloidal gold-labeled streptococcus G protein or colloidal gold-labeled staphylococcus aureus protein A; when the gold label pad is fixed with the colloidal gold labeled streptococcus G protein, the quality control line is coated with staphylococcus aureus protein A; when the gold label pad is fixed with the colloidal gold labeled staphylococcus aureus protein A, the quality control line is coated with streptococcus G protein; the gold label pad is connected with the end part of the nitrocellulose membrane in an overlapping way; the recombinant echinococcus antigen comprises recombinant echinococcus antigen Em18, recombinant echinococcus antigen EgCystatin and recombinant echinococcus antigen AgB 1.
3. The echinococcus antibody immunochromatographic test strip of claim 2, further comprising a sample pad, a water-absorbing pad and a bottom plate; the sample pad, the gold label pad, the nitrocellulose membrane and the absorbent pad are sequentially connected to the bottom plate in an end overlapping manner, wherein two ends of the nitrocellulose membrane are arranged at positions close to the bottom plate in the overlapping manner; the distance from the detection line group to the gold mark pad is shorter than the distance from the quality control line to the gold mark pad; the sample pad is a blood filtering membrane; the sample to be tested is selected from one or more of serum, plasma, whole blood, tissue fluid, urine, saliva and feces.
4. An echinococcus antibody immunochromatographic test strip which comprises a shell and the echinococcus antibody immunochromatographic test strip of claim 3; the casing is provided with:
a registration recording area for recording information including sample information, a detection date, and a detection result;
the observation window is used for observing the immunochromatography condition of the test strip arranged in the shell;
and the sample adding hole is used for adding a sample to the test strip arranged in the shell.
5. The echinococcus antibody immunochromatography detection card of claim 4, wherein the case is further provided with a protruding point region for holding the immunochromatography detection device; the housing includes a first end and a second end; the projection point area, the registration recording area, the observation window, and the sample application hole are sequentially provided between the first end portion and the second end portion.
6. The echinococcus antibody immunochromatography detection card of claim 4, wherein a test strip positioning plate and a test strip side positioning column are provided at the bottom of the inner surface of the case; the test strip side positioning columns are arranged on two sides of the test strip positioning plate; the top of the inner surface of the shell is provided with a test strip top positioning column, so that the test strip top positioning column can be pressed on the test strip positioning plate.
7. The echinococcus antibody immunochromatographic detection card of claim 4, wherein the shell comprises an upper shell and a lower shell; a clamping shell positioning column is arranged on the inner surface of the upper shell; the inner surface of the lower shell is provided with a clamping shell positioning hole in a matching mode, the clamping shell positioning hole is used for being inserted into the clamping shell positioning column and connecting the upper shell and the lower shell, and therefore the shells are in a closed state.
8. The echinococcus antibody immunochromatography detection card of claim 7, wherein the number of the card shell positioning holes and the number of the card shell positioning pillars are the same, and are 4-12.
9. The echinococcus antibody immunochromatographic detection card of claim 4, wherein the application hole is in a funnel shape with a wide top and a narrow bottom, and faces the sample pad of the test strip disposed in the case.
10. The echinococcus antibody immunochromatography detection card of claim 4, wherein the detection line and the quality control line can be observed through the observation window.
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