应用专一性寡核苷酸微阵列可靠地检测结核分枝杆菌利福平耐药性

DNA微阵列代表聚合酶链反应产物诊断测序的发展方向 .根据结核分枝杆菌rpoB基因利福平抗药性决定区域内点突变及其它重排的特征 .研制一种快速地鉴定结核分枝杆菌利福平耐药菌株的中等密度微阵列方法 .利福平抗药性通过使荧光标记扩增遗传物质与微阵列杂交测定 .检测5 3株利福平耐药结核分枝杆菌和 15株利福平敏感结核分枝杆菌 .微阵列方法的检测结果与药物敏感性试验和DNA测序结果完全一致 .临床标本PCR扩增后仅 1 5h可检出利福平耐药临床分离株 .表明寡核苷酸微阵列是高效的、专一性的方法 ,可作为检测利福平抗药性的快速方法以弥补传统培养方法的不足

Reliable Detection of Rifampin-Resistance of Mycobacterium tuberculosis Strains by Using a Specialized Oligonucleotide Microarray

A moderate density oligonucleotide microarray that could rapidly identify Mycobacterium tuberculosis rifampin resistant strains was developed. The method was based on the detection of point mutations and other rearrangements in the rpoB gene region determining rifampin resistance. Rifampin resistance was determined by hybridizing fluorescently labeled, amplified genetic material generated from bacterial colonies to the array. Fifty three rifampin resistant M.tuberculosis and fifteen rifampin susceptible M.tuberculosis were tested. The results were concordant with those based on culture drug susceptibility testing and sequencing. Rifampin resistant clinical isolates were detected in as little as 1.5 hours after PCR amplification with visual results. It was demonstrated that oligonucleotide microarray was efficient, specialized technique to implement and could be used as a rapid technique for detecting rifampin resistance to complement standard culture based method.