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一种用于结核分枝杆菌对异烟肼、利福平及链霉素耐药性诊断的高强度绿色荧光蛋白荧光噬菌体方法的评估

Evaluation of a High-Intensity Green Fluorescent Protein Fluorophage Method for Drug- Resistance Diagnosis in Tuberculosis for Isoniazid, Rifampin, and Streptomycin.

作者信息

Yu Xia, Gu Yunting, Jiang Guanglu, Ma Yifeng, Zhao Liping, Sun Zhaogang, Jain Paras, O'Donnell Max, Larsen Michelle, Jacobs William R, Huang Hairong

机构信息

National Clinical Laboratory on Tuberculosis, Beijing Key Laboratory for Drug-Resistant Tuberculosis Research, Beijing Chest Hospital, Beijing Tuberculosis and Thoracic Tumor Institute, Capital Medical University Beijing, China.

Department of Microbiology and Immunology, Albert Einstein College of Medicine Bronx, NY, USA.

出版信息

Front Microbiol. 2016 Jun 17;7:922. doi: 10.3389/fmicb.2016.00922. eCollection 2016.

Abstract

A novel method for detecting drug resistance in Mycobacterium tuberculosis using mycobacteriophage Φ (2) GFP10 was evaluated with clinical isolates. The phage facilitates microscopic fluorescence detection due to the high expression of green fluorescence protein which also simplifies the operative protocol as well. A total of 128 clinical isolates were tested by the phage assay for isoniazid (INH), rifampin (RIF), and streptomycin (STR) resistance while conventional drug susceptibility test, by MGIT960, was used as reference. The sensitivities of Φ (2) GFP10 assay for INH, RIF, and STR resistance detection were 100, 98.2, and 89.3%, respectively while their specificities were 85.1, 98.6, and 95.8%, respectively. The agreement between phage and conventional assay for detecting INH, RIF, and STR resistance was 92.2, 98.4, and 93.0%, respectively. The Φ (2) GFP10-phage results could be available in 2 days for RIF and STR, while it takes 3 days for INH, with an estimated cost of less than $2 to test all the three antibiotics. The Φ (2) GFP10-phage method has the potential to be a valuable, rapid and economical screening method for detecting drug-resistant tuberculosis.

摘要

采用分枝杆菌噬菌体Φ(2)GFP10检测结核分枝杆菌耐药性的新方法已通过临床分离株进行评估。该噬菌体因绿色荧光蛋白的高表达而便于显微镜荧光检测,这也简化了操作流程。通过噬菌体检测法对128株临床分离株进行异烟肼(INH)、利福平(RIF)和链霉素(STR)耐药性检测,同时以MGIT960法进行的传统药敏试验作为对照。Φ(2)GFP10检测法对INH、RIF和STR耐药性检测的灵敏度分别为100%、98.2%和89.3%,其特异性分别为85.1%、98.6%和95.8%。噬菌体检测法与传统检测法在检测INH、RIF和STR耐药性方面的一致性分别为92.2%、98.4%和93.0%。RIF和STR的Φ(2)GFP10噬菌体检测结果2天即可得出,INH则需要3天,检测三种抗生素的估计成本不到2美元。Φ(2)GFP10噬菌体法有可能成为一种有价值、快速且经济的耐药结核病筛查方法。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d5b3/4911409/e5b53c0acd94/fmicb-07-00922-g0001.jpg

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