一种简单经济的室内噬菌体技术，可直接在去污痰液样本中快速检测结核分枝杆菌对利福平、异烟肼、乙胺丁醇、链霉素和环丙沙星的耐药性。

A simple and economical in-house phage technique for the rapid detection of rifampin, isoniazid, ethambutol, streptomycin, and ciprofloxacin drug resistance in Mycobacterium tuberculosis, directly on decontaminated sputum samples.

机构信息

Department of Microbiology, Immunology and Molecular Biology, Intermediate Referral Laboratory for Mycobacteriology, Choithram Hospital and Research Centre, Manik Bagh Road, Indore, MP 452014, India.

出版信息

Int J Infect Dis. 2012 May;16(5):e332-6. doi: 10.1016/j.ijid.2011.12.016. Epub 2012 Feb 26.

DOI:10.1016/j.ijid.2011.12.016

PMID:22369736

Abstract

OBJECTIVES

The early detection of drug resistance would be a boon for TB control programs. The aim of the present study was to set up a rapid phage assay for the testing of drug susceptibility of Mycobacterium tuberculosis to rifampin, isoniazid, ethambutol, streptomycin, and ciprofloxacin, directly on decontaminated sputum samples.

METHODS

Mueller-Hinton broth was used instead of 7H9 broth to make the method more economical. Vancomycin and polymyxin B were added to the concentrated sputum samples to reduce the bacterial contamination. The phage assay on decontaminated sputum samples was compared with the proportion method using M. tuberculosis isolates from the same sputum samples.

RESULTS

Phage assay results were available within 48h for rifampin and streptomycin and within 72h for all the other drugs. In contrast the proportion method required 4-6 weeks from the primary cultures. The sensitivity of the phage assay was in the range of 93% to 100% and specificity in the range of 96% to 100% for all the drugs tested. The interpretation of results was possible for 334 of the 370 (90.3%) acid-fast bacillus (AFB) smear-positive sputum samples by the phage assay.

CONCLUSIONS

The phage assay for the detection of drug resistance on direct decontaminated sputum samples is economical, easy to perform, and rapid.

摘要

目的

耐药性的早期检测将是结核病控制项目的福音。本研究的目的是建立一种快速噬菌体检测法，直接检测结核分枝杆菌对利福平、异烟肼、乙胺丁醇、链霉素和环丙沙星的药物敏感性，用于已去污的痰标本。

方法

本研究使用 Mueller-Hinton 肉汤代替 7H9 肉汤，以降低成本。浓缩的痰标本中加入万古霉素和多粘菌素 B 以减少细菌污染。将去污痰标本的噬菌体检测法与来自同一痰标本的结核分枝杆菌分离株的比例法进行比较。

结果

噬菌体检测法对利福平、链霉素的结果可在 48 小时内得到，对所有其他药物的结果可在 72 小时内得到。相比之下，比例法需要从初次培养开始 4-6 周。噬菌体检测法的灵敏度范围为 93%至 100%，特异性范围为 96%至 100%，适用于所有测试的药物。噬菌体检测法可对 370 份酸染色阳性痰标本中的 334 份（90.3%）进行结果解读。