Method for detecting rifampin drug resistance of mycobacterium tuberculosis by using pyrosequencing technology
A technology of Mycobacterium tuberculosis and pyrosequencing, which is applied in the directions of microorganism-based methods, microbial determination/inspection, biochemical equipment and methods, etc. Many times, etc., to achieve the effect of shortening the detection time, reducing the number of reactions and the amount of detection reagents
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[0026] Since the mutations of the rpoB gene are mainly concentrated in the RRDR region, and there are many mutation sites in the RRDR region, the pyrosequencing detection method of the present invention is mainly aimed at the RRDR region of the rpoB resistance gene of rifampin. The present invention provides a method for detecting the resistance of Mycobacterium tuberculosis to rifampicin. The rpoB gene is first amplified by PCR, and then the PCR amplified product is subjected to pyrosequencing using SQA mode to determine whether the rpoB gene has mutations Site, where the nucleotide loading sequence is AGCT.
[0027] Pyrosequencing technology
[0028] Pyrosequencing (Pyrosequencing) technology is a brand-new DNA sequence analysis technology that can quickly and accurately determine a short target fragment. This technique does not require electrophoresis, and DNA fragments do not require fluorescent labeling, and the operation is extremely simple. Because the pyrosequencing tech...
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