Method for identifying rifampicin heterogeneity drug resistance of mycobacterium tuberculosis based on fluorescein flow cytometry

Abstract

The invention relates to the field of drug sensitivity detection, in particular to a method for identifying rifampicin heterogeneity drug resistance of mycobacterium tuberculosis based on fluoresceinflow cytometry. The method comprises the following steps: S1, a working solution is prepared; S2, analysis bacterial solutions are treated for 10 days or 14 days by using a substitution solution and arifampicin working solution respectively; S3, the treated bacterial solutions are dyed respectively; and S4, the dyed bacterial solutions are detected respectively, an average fluorescence intensityvalue of each bacterial solution is recorded, a sensitivity index of each bacterial solution is calculated, and the sensitivity of each bacterial solution to rifampicin is judged according to the sensitivity index: when the sensitivity index is higher than a critical value, the detected mycobacterium tuberculosis is heterogeneity drug-resistant mycobacterium tuberculosis or drug-resistant mycobacterium tuberculosis, when the sensitivity index is lower than the critical value, the the detected mycobacterium tuberculosis is a sensitive bacterium, and the critical value range is 0.48 or 0.37. Themethod is relatively high in accuracy, a detection result of rifampicin heterogeneity drug resistance of the mycobacterium tuberculosis can be rapidly obtained within 10 days, and the accuracy, sensitivity and specificity are high.

Description

technical field [0001] The invention relates to the field of drug sensitivity detection, in particular to a method for identifying heterogeneous resistance to rifampicin of mycobacterium tuberculosis by flow cytometry based on fluorescein. Background technique [0002] Flow cytometry has the advantages of high throughput, multi-parameters, and high efficiency, and is increasingly used in microbial drug susceptibility testing. The essence of detecting bacterial drug sensitivity is to monitor the activity of bacteria after drug treatment. Flow cytometry reflects the activity of bacteria by monitoring the difference in fluorescence value after the interaction between bacteria and fluorescent dyes in different states. The viability of sensitive bacteria decreases after being treated with antibiotics, while the viability of drug-resistant bacteria does not change. If microorganisms with different vigor or functional states combine with specific fluorescent dyes to emit fluorescen...

AI Technical Summary

Problems solved by technology

Although the current drug resistance detection technology has become mature, for example, the traditional proportional method DST is a reliable and sensitive method for detecting drug resistance, but to detect and obtain the heterogeneous drug resistance of pathogenic bacteria, the current technology For the case, it must rely on genetic testing technology

However, there are defects in genetic testing that can only detect drug-resistant gene mutations that are clearly related to the phenotype, and cannot identify unspecified drug-resistant genes, which will eventually affect the detection accuracy and reduce the clinical reference value of the test results.

[0005] At present, no other more accurate and convenient technical means have been effectively applied to the detection of heterogeneous drug resistance of Mycobacterium tuberculosis.

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