Rapid Detection of Isoniazid Resistance in Mycobacterium tuberculosis Isolates by Use of Real-Time-PCR-Based Melting Curve Analysis

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	Rapid Detection of Isoniazid Resistance in Mycobacterium tuberculosis Isolates by Use of Real-Time-PCR-Based Melting Curve Analysis
	Hu, Siyu ; Li, Guoli ; Li, Hui ; Liu, Xiaoli ; Niu, Jianjun ; Quan, Shengmao ; Wang, Feng ; Wen, Huixin ; Xu, Ye ; Li, Qingge ; Wang F(王凤) ; Xu Y(许晔) ; Li QG(李庆阁)
刊名	http://dx.doi.org/10.1128/JCM.03395-13
	2014
关键词	RIFAMPIN RESISTANCE MOLECULAR-DETECTION TREATMENT OUTCOMES ASSAY STRAINS HETERORESISTANCE SUSCEPTIBILITY MUTATIONS EVOLUTION INHA
英文摘要	Important National Science Technology [2008ZX10003-004]; Xiamen Science and Technology [3502Z20113009]; The MeltPro TB/INH assay, recently approved by the Chinese Food and Drug Administration, is a closed-tube, dual-color, melting curve analysis-based, real-time PCR test specially designed to detect 30 isoniazid (INH) resistance mutations in katG position 315 (katG 315), the inhA promoter (positions -17 to -8), inhA position 94, and the ahpC promoter (positions -44 to -30 and -15 to 3) of Mycobacterium tuberculosis. Here we evaluated both the analytical performance and clinical performance of this assay. Analytical studies with corresponding panels demonstrated that the accuracy for detection of different mutation types (10 wild-type samples and 12 mutant type samples), the limit of detection (2 x 103 to 2 x 104 bacilli/ml), reproducibility (standard deviation [SD],< 0.4 degrees C), and the lowest heteroresistance level (40%) all met the parameters preset by the kit. The assay could be run on five types of real-time PCR machines, with the shortest running time (105 min) obtained with the LightCycler 480 II. Clinical studies enrolled 1,096 clinical isolates collected from three geographically different tuberculosis centers, including 437 INH-resistant isolates and 659 INH-susceptible isolates characterized by traditional drug susceptibility testing on Lowenstein-Jensen solid medium. The clinical sensitivity and specificity of the MeltPro TB/INH assay were 90.8% and 96.4%, respectively. DNA sequencing analysis showed that, except for the 5 mutants outside the detection range of the MeltPro assay, a concordance rate between the two methods of 99.1% (457/461) was obtained. Among the 26 mutation types detected, katG S315T (AGC -> ACC), inhA -15C -> T, katG S315N (AGC -> AAC), and ahpC promoter -10C -> T accounted for more than 90%. Overall, the MeltPro TB/INH assay represents a reliable and rapid tool for the detection of INH resistance in clinical isolates.
语种	英语
出版者	AMER SOC MICROBIOLOGY
内容类型	期刊论文
源URL	[http://dspace.xmu.edu.cn/handle/2288/93614]
专题	医学院－已发表论文
推荐引用方式 GB/T 7714	Hu, Siyu,Li, Guoli,Li, Hui,et al. Rapid Detection of Isoniazid Resistance in Mycobacterium tuberculosis Isolates by Use of Real-Time-PCR-Based Melting Curve Analysis[J]. http://dx.doi.org/10.1128/JCM.03395-13,2014.
APA	Hu, Siyu.,Li, Guoli.,Li, Hui.,Liu, Xiaoli.,Niu, Jianjun.,...&李庆阁.(2014).Rapid Detection of Isoniazid Resistance in Mycobacterium tuberculosis Isolates by Use of Real-Time-PCR-Based Melting Curve Analysis.http://dx.doi.org/10.1128/JCM.03395-13.
MLA	Hu, Siyu,et al."Rapid Detection of Isoniazid Resistance in Mycobacterium tuberculosis Isolates by Use of Real-Time-PCR-Based Melting Curve Analysis".http://dx.doi.org/10.1128/JCM.03395-13 (2014).