A Fluorescein-Based Flow Cytometry Method for Identification of Rifampicin Heterogeneous Drug Resistance in Mycobacterium tuberculosis

Abstract

The present invention relates to the field of drug sensitivity detection, specifically a method for identifying heterogeneous resistance to rifampicin of Mycobacterium tuberculosis by fluorescein-based flow cytometry. The steps include: S1, preparation of working solution; S2, respectively Treat the analyzed bacterial solution with the replacement solution and rifampicin working solution for 10 days or 14 days; S3: stain the treated bacterial solution separately; S4: test the stained bacterial solution separately, and record the average value of each bacterial solution Fluorescence intensity value, and calculate its sensitivity index, and judge its sensitivity to rifampicin according to the sensitivity index: when the sensitivity index is greater than the critical value, it is judged that the detected Mycobacterium tuberculosis is heterogeneous drug-resistant Mycobacterium tuberculosis or resistant Mycobacterium tuberculosis; when the sensitivity index is lower than the critical value, it is judged as a sensitive bacteria; the critical value range is 0.48 or 0.37. The method of the invention has high accuracy, can quickly obtain the detection result of rifampicin heterogeneity drug resistance of Mycobacterium tuberculosis within 10 days, and has high accuracy, sensitivity and specificity.

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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