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Immunochromatographic strip for detecting viscerotropic leishmania infection and diagnosing kala-azar

A leishmania and immunochromatography technology, applied in the field of bioengineering, can solve the problems of complex methods and low detection rate for detecting visceral leishmania infection and diagnosing kala-azar disease, and achieves rapid sample processing and detection methods. Simple, sensitive and specific effects

Inactive Publication Date: 2012-07-18
STATION OF VIRUS PREVENTION & CONTROL CHINA DISEASES PREVENTION & CONTROL CENT
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] The purpose of the present invention is to provide a kind of immunochromatographic test strip and its preparation method for detecting the infection of visceraphim Leishmania and diagnosing kala-azar, and described this immunochromatographic test strip and its preparation method will solve the problems in the prior art. The method for detecting Leishmania viscerophila infection and diagnosing kala-azar disease is complicated and has low detection rate technical problems

Method used

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  • Immunochromatographic strip for detecting viscerotropic leishmania infection and diagnosing kala-azar
  • Immunochromatographic strip for detecting viscerotropic leishmania infection and diagnosing kala-azar
  • Immunochromatographic strip for detecting viscerotropic leishmania infection and diagnosing kala-azar

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0035] Example 1 Immunochromatography test strip

[0036] Such as figure 1 and 2 As shown, an immunochromatographic test strip for detecting visceral Leishmania infection and diagnosing kala-azar, comprising a sample pad 1, a gold standard pad 2 tightly connected to the sample pad 1 containing a colloidal gold-labeled probe, A cellulose film 3 tightly connected to the gold standard pad 2, and a water-absorbent pad 4 closely connected to the cellulose film 3, the water-absorbent pad 4 is far away from the gold standard pad 2, and on the fiber A quality control line 6 is set near the end of the water-absorbing pad 4 on the plain film 3, and a test line 5 is set on the cellulose film 3 between the quality control line 6 and the gold standard pad 2, and the test line 5 contains pro-viscera. The parasite soluble antigen of Shmania, the quality control line 6 contains an antibody that specifically binds to the colloidal gold-labeled probe.

[0037] Further, the parasite soluble a...

Embodiment 2

[0042] Embodiment 2 Detection principle and result judgment

[0043] Add the sample serum or whole blood on the blood filter membrane sample pad 1 during the measurement, drop a drop (about 50 μL) of sample diluent after 1 minute, and the diluent drives the sample to press the sample pad 1, gold standard pad 2, and nitrocellulose membrane 3 1. Move in the direction of the absorbent pad 4. When flowing through the gold standard pad 2, the colloidal gold labeling probe on the gold standard pad 2 is redissolved, and drives it to move to the nitrocellulose membrane 3 and the absorbent pad 4. The colloidal gold-labeled probe (the embodiment of the present invention is Staphylococcus aureus protein A) can combine with the antibody or antibody subclass in the sample to form an immune complex. When the immune complex flows to the detection line 5, if there are antibodies to be tested or antibody subclasses (anti-Leishmania antigen antibodies) in the specimen, they will be captured by ...

Embodiment 3

[0045] Example 3 Preparation of Immunochromatographic Test Strips

[0046] 1. Preparation of parasite soluble antigen against Leishmania donovani

[0047] The promastigotes of Leishmania donovani were inoculated in 199 medium and cultured in an incubator at 22 °C. The protozoa was in the logarithmic growth phase, and the protozoan density was about 10 6 At 4°C, centrifuge at 3000g for 15 minutes to collect promastigotes, discard the supernatant, wash the pellet with PBS three times in the same way, add 4 times the volume of PBS according to the volume of promastigotes, and store in liquid nitrogen and In a water bath at 37°C, freeze and thaw repeatedly 5 times, then ultrasonically pulverize 3 times in an ice bath, centrifuge at 18000g, 4°C for 20min, and the supernatant is the soluble antigen.

[0048] 2. Staphylococcus aureus protein A (SPA) can be purchased

[0049] 3. Preparation of immunocolloidal gold probes and gold standard pads

[0050] Prepare Staphylococcus aureus...

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Abstract

The invention discloses an immunochromatographic strip for detecting viscerotropic leishmania infection and diagnosing kala-azar, which comprises a sample pad, a conjugate pad which is closely connected to the sample pad and contains a colloidal gold-labeled probe, a cellulose membrane closely connected with the conjugate pad and a water absorption pad closely connected to the cellulose membrane. The water absorption pad is far away from the conjugate pad, a quality control line is arranged at one end of the cellulose membrane, which is close to the water absorption pad, a detection line is positioned on the cellulose membrane between the quality control line and the conjugate pad, the detection line contains soluble antigen of the viscerotropic leishmania, and the quality control line contains antibodies specifically binding to the colloidal gold-labeled probe. The invention further provides a kit containing the immunochromatographic strip and a preparation method for the immunochromatographic strip. The immunochromatographic strip has the advantages of being simple and convenient to use, high in sensitivity and specificity, fast in detection, and simultaneously applicable to clinical and field use for human, livestock and wild animals.

Description

technical field [0001] The invention relates to the field of bioengineering, in particular to a detection tool for Leishmania viscera parasitic diseases, in particular to an immunochromatographic test strip for detecting Leishmania viscera infection and diagnosing kala-azar and a preparation method thereof. Background technique [0002] The life cycle of Leishmania spp. has two stages: promastigote and amastigote. The former parasitizes in the digestive tract of arthropods (sandflies), while the latter parasitizes in the cells of mammals or reptiles and spreads through sandflies. The classification of Leishmania is complex, and visceral leishmaniasis or kala-azar is parasitized by Leishmania donovani or L.infantum / L.chagasi A parasitic disease caused by the lymphoid-macrophage system of the human body and transmitted by the bite of the vector sandfly. The blood of kala-azar patients contains anti-Leishmania antibodies that can specifically bind to the infected Leishmania a...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/569G01N33/577G01N33/68A61K39/008A61P33/02
CPCY02A50/30
Inventor 石锋高春花汪俊云杨玥涛丁丹
Owner STATION OF VIRUS PREVENTION & CONTROL CHINA DISEASES PREVENTION & CONTROL CENT
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