Immunoassay method and kit for assaying mycobacterium tuberculosis from biological samples

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...

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 other 20% Most tuberculosis patients present with extrapulmonary tuberculosis, and sputum samples cannot be obtained, so no clear bacteriological evidence can be obtained. They can only rely on other methods for auxiliary diagnosis, or tentative anti-tuberculosis treatment. So far, new drugs suitable for bacterial negative and extrapulmonary tuberculosis are needed. Bacteriological detection method

However, the above methods all use DNA as a template, and the stability of DNA makes it impossible to distinguish live bacteria from dead bacteria in the PCR test results, resulting in false positive results.

[0026] However, compared with sputum smear testing, molecular testing is still more complicated, and it is also mainly based on sputum samples. At present, there is no relevant research on the detection results of molecular diagnostic methods for blood samples.

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