Immunoassay method and kit for assaying mycobacterium tuberculosis from biological samples

A technology of Mycobacterium tuberculosis and an immune detection method, applied in the field of medical immunodiagnostics, can solve the problems of high complexity of molecular detection, inability to obtain higher detection rate specificity, inability to obtain bacteriological evidence, etc.

Pending Publication Date: 2016-07-13
肖乐义
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0011] First, the detection is based on the patient's sputum sample. Factors such as the presence or absence of the patient's sputum, whether the collection method is correct, whether the quality is good or bad, and whether it is sent for inspection in a timely manner will directly affect the test results, resulting in the inability to obtain bacteriological tests based on sputum samples. With higher detection rate and better specificity, new detection methods are needed to replace traditional sputum detection
[0012] Second, pulmonary tuberculosis patients account for 80% of all tuberculosis patients, and nearly 50% of them are patients with negative sputum bacteria. The traditional bacteriological detection of sputum bacteria will inevitably lead to missed diagnosis of this part of the population, and other methods are needed to assist diagnosis; the o...

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  • Immunoassay method and kit for assaying mycobacterium tuberculosis from biological samples
  • Immunoassay method and kit for assaying mycobacterium tuberculosis from biological samples
  • Immunoassay method and kit for assaying mycobacterium tuberculosis from biological samples

Examples

Experimental program
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Embodiment 1

[0048] Detection and Identification of Mycobacterium tuberculosis in Example 1 Plasma

[0049] (1) Collect 5-10ml of the subject's blood (anticoagulation, EDTA anticoagulation can help to detect Mycobacterium tuberculosis and then do molecular detection and drug resistance test);

[0050] (2) Centrifuge at 500-1500 rpm for 5-10 minutes (it can also allow blood cells to sink naturally and then separate plasma and blood cells);

[0051] (3) Based on microporous membrane (the material of microporous membrane can be any suitable material, such as polycarbonate membrane, cellulose ester membrane, polyamide membrane, polysulfone membrane, fluorine-containing material membrane, polyester membrane Membranes, polyolefin membranes, inorganic material membranes, etc., preferably polycarbonate membranes.) microfluidic filtration device to filter and enrich Mycobacterium tuberculosis;

[0052] (4) Plasma 2-10ml, take 2ml as an example to illustrate the method and process;

[0053] (5) Ad...

Embodiment 2

[0062] Example 2 Detection and identification of mycobacterium tuberculosis in cerebrospinal fluid

[0063] (1) 0.5-2ml of freshly collected cerebrospinal fluid, to avoid cell damage, decomposition of glucose and other substances, bacterial dissolution, etc. due to the specimen being placed for too long, and the specimen should try to avoid coagulation and mixing into the blood;

[0064] (2) Membrane-based microfluidic filtration device to filter and enrich Mycobacterium tuberculosis;

[0065] (3) Cerebrospinal fluid 0.5-2ml, take 0.5ml as an example to illustrate the method and process;

[0066] (4) Add 0.5ml of cerebrospinal fluid to 3.5ml of normal saline or PBS, and mix well;

[0067] (5) Add 8ml of 6% sodium hypochlorite, mix well, and let stand for 5-20 minutes;

[0068] (6) Add 48ml of 95 ethanol (containing 1% TritonX-100), mix well, and let stand for 5-15 minutes;

[0069] (7) Add the above-mentioned liquid into a microfluidic device or filter and enrich Mycobacter...

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Abstract

The invention discloses a method and a kit for quickly assaying mycobacterium tuberculosis from blood or cerebrospinal fluid by means of fluorescence staining.Particularly, anti-mycobacterium tuberculosis antibodies, especially monoclonal or polyclonal antibodies of ESAT-6 and CFP-10 antigens in RD-1 (region of difference-1) are used for assaying the mycobacterium tuberculosis from biological samples enriched by a microporous membrane-based enrichment device through an indirect or direct immunization method or an SPA method.The method has the advantages of simplicity in operation, high sensitivity, high specificity and capability of distinguishing characteristics of pathogenic mycobacterium tuberculosis and non-pathogenic mycobacterium tuberculosis caused by Bacillus Calmette-Guerin vaccination, can be used for pathogenic detection of tuberculosis diseases and early rapid clinical diagnosis and is a specific tuberculosis etiology diagnostic method.

Description

technical field [0001] The invention belongs to the technical field of medical immunodiagnosis, in particular, the invention relates to an immunodetection method and a kit for detecting Mycobacterium tuberculosis in biological samples. Background technique [0002] Tuberculosis is a chronic infectious disease caused by a single pathogen, Mycobacterium tuberculosis. In recent years, due to factors such as the increase of population mobility, the emergence of drug-resistant tuberculosis, and the double infection of tuberculosis and HIV, tuberculosis has made a comeback and has once again become a major public health and social problem that seriously endangers the health of the people. According to the report of the World Health Organization (WHO), it is currently estimated that about 2 billion people in the world are infected with Mycobacterium tuberculosis, with 9 million new cases in 2013. my country is a country with a high burden of tuberculosis. The number of tuberculosi...

Claims

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Application Information

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IPC IPC(8): G01N33/569
CPCG01N33/5695G01N2333/35
Inventor 肖乐义张贺秋米明仁冯晓燕陈立敏杨勤英谭印成陈凤华
Owner 肖乐义
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