Mycobacterium tuberculosis complex rapid detection kit and preparation method thereof

Abstract

The invention discloses a mycobacterium tuberculosis complex rapid detection kit, which comprises an identification reagent, and the identification reagent comprises at least one pair of ten pairs ofprimers and at least one of ten probes. The invention also provides a preparation method of the rapid detection kit for the mycobacterium tuberculosis complex. The preparation method comprises the following steps of: S1, designing primers and probes according to gene sequences of the mycobacterium tuberculosis complex compared in a gene pool; S2, constructing a kit program of THE mycobacterium tuberculosis complex; S3, testing the specificity and sensitivity of the kit program; S4, carrying out repeated tests on the established kit program for many times; S5, preparing a standard target gene pool; and S6, constructing a specific detection kit with high sensitivity. The invention has the beneficial effects that by utilizing the kit based on the specific primer pair and the probe provided bythe invention, mycobacterium tuberculosis can be rapidly and comprehensively detected and identified, and possibility is provided for subsequent comprehensive detection; the sensitivity test, the specificity test and the repeatability test are utilized, and meanwhile, the CT difference value is compared, so that rapid and accurate detection is achieved.

Description

technical field [0001] The invention relates to the field of detection kits for Mycobacterium tuberculosis, in particular to a rapid detection kit for Mycobacterium tuberculosis complex and a preparation method thereof. Background technique [0002] Tuberculosis is one of the major infectious diseases in the world, and my country is a big tuberculosis country in the world and it continues to grow. According to statistics, one out of every three people has Mycobacterium tuberculosis, and the number of infected people has far exceeded 400 million. , of which 90%-98% are in the incubation period. The disease is a chronic infectious disease caused by Mycobacterium tuberculosis (M.tuberculosis), Mycobacterium microti (M.microti), Mycobacterium bovis (M.bovis), etc. Organs, mostly the lungs. [0003] As early as 17,000 years ago, some scholars discovered Mycobacterium tuberculosis in the remains of bison, which is the earliest origin of tuberculosis found so far [Lu Zhiming. The ...

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Problems solved by technology

[0006] The existing nucleic acid detection methods using detection kits to detect Mycobacterium tuberculosis mostly focus on the genetic design of a certain type or several types of Mycobacterium tuberculosis, but lack of comparison and elimination of other Mycobacterium tuberculosis, which has certain advantages. limitations; such as the invention is a kit and method for identifying mycobacterial strains (application number: CN201510812703.9), comprising the following steps: (1) design and synthesis of primers and probes; (2) PCR Reaction components, (3) PCR amplification; (4) melting curve analysis

Although dual-label and dual-specific probes were designed to detect 12 clinical mycobacterial species, which solved the problem of relatively few species of Mycobacterium tuberculosis compared in genetic design, but only dual-specific probes were used In the detection of multiple mycobacterium tuberculosis mixed, due to the masking of other mycobacterium tuberculosis or the disturbance of color development, there will be errors in probe detection; When Bacillus is amplified, its marker characteristics may not necessarily increase with the amplification, and it is prone to underreporting

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