State Key Laboratory of Genetic Engineering, Institute of Genetics, School of Life Science, Fudan University, Shanghai 200433, China.
Microbiol Res. 2011 Feb 20;166(2):121-8. doi: 10.1016/j.micres.2010.02.001.
Drug resistance remains a serious threat to tuberculosis control worldwide. As one of the important first-line antitubercular agents, resistance to streptomycin (SM) and its derivatives has increased in recent years and has become one of the characteristics of extensively drug-resistant tuberculosis (XDR-TB). A novel rapid assay to screen for rpsL gene mutations associated with SM resistance in Mycobacterium tuberculosis (M. tuberculosis), was developed using high-resolution melting (HRM) analysis. The HRM results of 134 SM-resistant clinical isolates and 20 SM-susceptible clinical isolates of M. tuberculosis for rpsL gene mutations were perfectly matched with those of DNA sequencing. SM resistance was highly associated with rpsL mutations in M. tuberculosis. HRM technique represented an inexpensive, highly sensitive and high-throughput method to facilitate the screening of large numbers of clinical samples for epidemiological studies of drug-resistance of M. tuberculosis, especially in developing countries.
耐药性仍然是全球结核病控制的严重威胁。作为重要的一线抗结核药物之一，近年来链霉素（SM）及其衍生物的耐药性有所增加，已成为广泛耐药结核病（XDR-TB）的特征之一。本研究利用高分辨率熔解（HRM）分析，开发了一种新型快速检测结核分枝杆菌（M. tuberculosis）中 rpsL 基因突变与 SM 耐药性相关的方法。HRM 结果与 DNA 测序结果完全一致，共检测了 134 株 SM 耐药临床分离株和 20 株 SM 敏感临床分离株的 rpsL 基因突变。SM 耐药性与 M. tuberculosis rpsL 突变高度相关。HRM 技术代表了一种廉价、高敏感、高通量的方法，可用于筛选大量临床样本，以进行 M. tuberculosis 耐药性的流行病学研究，特别是在发展中国家。
