Human body intestinal canal flora detection parting and quantitative reagent kit

Abstract

The invention relates to a method for detecting, genotyping and quantifying human intestinal flora, consisting of universal primer PCR amplification and specific probe LDR. In the PCR amplification, the 16sr DNA fragment of selected bacteria genus is amplified by the designed universal primer synchronously; the specific LDR probe of each bacteria genus is utilized to achieve the goal of genotyping through ligation reaction; and LDR signal is detected by ABI Prism 377 type sequencer, and the detected fluorescence value is taken as the quantifying basis. Aiming at the universal primer of intestinal microflora, the detection method of the invention effectively amplifies all target intestinal microflora 16sr DNA sequence, thereby ensuring that each target sequence and the template of low copy therein can be detected, further ensuring the specificity of the specific probe of each bacteria genus, completely identifying each target sequence and realizing multiplex genotyping quantification.

Description

technical field [0001] The invention belongs to the field of microbial nucleic acid diagnosis, and in particular relates to a detection, typing and quantification kit of human intestinal flora. Background technique [0002] Bacteria in the intestinal tract play an important role in human growth and development, immune function, nutrition, drug metabolism, pathogenesis and health. The isolation, identification and quantification of intestinal bacteria are very important for studying the function of intestinal flora and the relationship with the body. At present, several methods for typing rRNA genes of intestinal bacteria have been established at home and abroad, but these methods generally have weaknesses such as unstable results, difficult operation, small throughput, and inability to quantify, which hinder research institutions and clinical units from identifying intestinal bacteria. Carry out effective scientific research and detection of the intestinal flora. [0003] ...

AI Technical Summary

Problems solved by technology

However, in most cases, a variety of bacteria are often mixed in the sample to be tested. At this time, selective identification medium must be used for differential counting. Various bacteria have different oxygen tolerance, nutritional requirements, antibiotic sensitivity and colony morphology. And the color characteristics constitute the basis of identification. At the same time, the antibiotics added in the culture medium will also have different degrees of inhibition on the bacteria themselves, and the specific culture medium is only relatively specific, so far there is no completely ideal selective culture For these reasons, although the traditional bacterial culture method can count viable bacteria, it is time-consuming, laborious, difficult to cultivate, and the count is not accurate. The identification basis of the traditional bacterial culture method is based on the morphology, physiology and biochemical characteristics of bacteria. , and these characteristics are only the specific performance of bacteria under special culture conditions,

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