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Lyme disease reagent kit and its lyme disease detecting method

A Lyme disease and kit technology, applied in the field of Lyme disease kits, can solve the problems of different antigenicity, different antigens that cannot be excluded from the cross of Lyme disease spirochetes, etc.

Inactive Publication Date: 2004-01-21
NO 261 HOSPITAL PLA
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Problems solved by technology

However, the ratio of its various antigens is determined by the composition of the bacteria itself. In fact, different antigens play different roles in diagnosis (that is, the antigenicity is different). At the same time, it cannot exclude Lyme spirochete and Other bacteria have cross-antigens

Method used

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Experimental program
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Embodiment Construction

[0018] The composition of kit of the present invention:

[0019] 1) Polystyrene microporous plate (Shenzhen No. 1 Chemical Factory).

[0020] 2) Enzyme-labeled mouse anti-human IgG antibodies were screened, and horseradish enzymes used in Concord Basic Research were selected, and 2 strains of mouse anti-human monoclonal antibodies were selected. The mixed antibodies were applied secondary antibodies, and the concentration used was 1:10000 (2D1 and 2D5) .

[0021] 3) Screening of P39 protein antigen coating concentration, the applied concentration is 145ng / ml. The antigen was successfully cloned using the BmpA or P39 gene of the Lyme spirochete strain isolated by my country itself, and was produced by high-efficiency expression in the form of natural protein in Escherichia coli.

[0022] 4) Reagents

[0023] A blocking solution

[0024] NaCl 8.0g Na 2 HPO 4 .12H 2 O 5.8g NaH 2 PO 4 2H 2 O 0.593g Casein2.5g Sucrose 50g Horse serum 22mL H 2 O 1000mL

[0025] B sample ...

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Abstract

The present invention is Lyme disease detecting reagent kit and its Lyme disease detecting method. The reagent kit includes P39 protein antigen coated carrier micropore plate, enzyme labeling antibody, substrate liquid, etc. It has the advantages of powerful and stable antigen immunogenicity and capability of mass production, and one set of reagent and several monoclonal mouse-to-human IgG antigens may be used to eliminate crossed display effect to result in clear background. The reagent kit is simple in operation, needs no special instrument and may be used in common hospital for wide epidemic disease survey and patient detection.

Description

field of invention [0001] The invention relates to a Lyme disease kit and a method for detecting Lyme disease. Background technique [0002] Serological detection methods for Lyme disease mainly use known antigens to detect specific antibodies against Lyme spirochete that may exist in patients. The most commonly used methods are indirect immunofluorescence assay (IFA), enzyme-linked immunosorbent assay (ELISA), western blotting, molecular biology method polymerase chain (PCR), tissue microscopy and pathogenic bacteria culture in vitro. [0003] So far, the laboratory diagnosis of Lyme disease has not been effectively resolved, and there is still no uniform reference standard globally. [0004] Traditional serological tests mostly use the whole Borrelia Lyme disease antigen as the standard antigen, and the cross-reactivity is relatively serious, such as enzyme-linked immunosorbent assay (ELISA). Indirect immunofluorescence assay (IFA) and enzyme-linked immunosorbent assay (...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/53G01N33/547G01N33/569G01N33/577
CPCY02A50/30
Inventor 贾月萍佟玉偘张晓芳陈昌松祝杨王东香
Owner NO 261 HOSPITAL PLA
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