检测艰难梭菌耐莫西沙星gyrA基因点突变的双重荧光PCR方法的建立和评价
Disease Surveillance（2021）
Abstract
目的 建立一种检测艰难梭菌耐莫西沙星gyrA基因点突变的双重荧光PCR方法.方法 设计针对艰难梭菌gyrA基因的特异性引物,并针对莫西沙星耐药株和敏感株的gyrA基因突变位点设计不同的TaqMan-MGB探针,优化可同时检测ATT、ACT突变点的双重荧光定量PCR方法,验证该方法的灵敏性、特异性和重复性,并进行应用评价.结果 该方法对莫西沙星耐药株和敏感株gyrA基因突变位点的检测下限分别可达4 copies/μL和5 copies/μL,对艰难梭菌以外其他常见肠道菌群的检测均为阴性.重复性检测结果显示,批间和批内变异系数均＜5％,该方法的检测结果与测序结果一致性高(k=0.98).结论 针对耐莫西沙星艰难梭菌gyrA基因点突变建立的双重荧光定量PCR方法,灵敏且具有很高的特异性和重复性,可有效鉴别艰难梭菌莫西沙星敏感株和耐药株,并可辅助识别流行株BI/NAP1/RT027的暴发.
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