Evaluation of the MeltPro TB/STR assay for rapid detection of streptomycin resistance in Mycobacterium tuberculosis. Issue 2 (March 2015)

• Record Type: Journal Article
• Authors: Zhang, Ting; Hu, Siyu; Li, Guoli; Li, Hui; Liu, Xiaoli; Niu, Jianjun; Wang, Feng; Wen, Huixin; Xu, Ye; Li, Qingge
• Is Part Of: Tuberculosis. Volume 95:Issue 2(2015)
• Page Start: 162
• Page End: 169
• Publication Date: 2015-03
• Subjects: 616.995
• DOI: 10.1016/j.tube.2014.12.004
• Languages: English
• ISSNs: 1472-9792

Abstract

<abstract xml:lang="en" abstract-type="author" id="abs0010"> <title id="sectitle0010">Summary</title> <sec> <p id="abspara0010">Rapid and comprehensive detection of drug-resistance is essential for the control of tuberculosis, which has facilitated the development of molecular assays for the detection of drug-resistant mutations in <italic>Mycobacterium tuberculosis</italic>. We hereby assessed the analytical and clinical performance of an assay for streptomycin-resistant mutations. MeltPro TB/STR is a closed-tube, dual-color, melting curve analysis-based, real-time PCR test designed to detect 15 streptomycin-resistant mutations in <italic>rpsL</italic> 43, <italic>rpsL</italic> 88, <italic>rrs</italic> 513, <italic>rrs</italic> 514, <italic>rrs</italic> 517, and <italic>rrs</italic> 905–908 of <italic>M</italic>. <italic>tuberculosis</italic>. Analytical studies showed that the accuracy was 100%, the limit of detection was 50–500 bacilli per reaction, the reproducibility in the form of <italic>T</italic><sub><italic>m</italic></sub> variation was within 1.0 °C, and we could detect 20% STR resistance in mixed bacterial samples. The cross-platform study demonstrated that the assay could be performed on six models of real-time PCR instruments. A multicenter clinical study was conducted using 1056 clinical isolates, which were collected from three geographically different healthcare units, including 709 STR-susceptible and 347 STR-resistant isolates characterized on Löwenstein–Jensen solid medium by traditional drug susceptibility testing. The results showed that the clinical sensitivity and specificity of the MeltPro TB/STR was 88.8% and 95.8%, respectively. Sequencing analysis confirmed the accuracy of the mutation types. Among all the 8 mutation types detected, <italic>rpsL</italic> K43R (AAG → AGG), <italic>rpsL</italic> K88R (AAG → AGG) and <italic>rrs</italic> 514 A → C accounted for more than 90%. We concluded that MeltPro TB/STR represents a rapid and reliable assay for the detection of STR resistance in clinical isolates.</p> </sec> </abstract>