摘要
目的分析南京市结核分枝杆菌(MTB)的耐药情况,评价荧光PCR熔解曲线法和PCR-线性杂交酶显色法检测结核分枝杆菌对利福平(RFP)和异烟肼(INH)耐药的应用价值。方法回顾性分析2017年1月-2021年3月南京市公共卫生医疗中心荧光PCR熔解曲线法和PCR-线性杂交酶显色法检测151份结核分枝杆菌临床分离株利福平和异烟肼耐药突变情况,以表型药敏试验结果为标准,评价以上2种分子生物学方法的诊断效能。结果151份结核分枝杆菌临床分离株中,6份为利福平单耐药株,9份为异烟肼单耐药株,43份为耐多药株。以表型药敏试验检测结果为标准,荧光PCR熔解曲线法检测利福平和异烟肼耐药性的敏感度、特异度、符合率以及κ值分别为100.00%和94.23%、96.08%和96.97%、97.35%和96.02%以及0.94和0.91。以表型药敏试验检测结果为标准,PCR-线性杂交酶显色法检测利福平和异烟肼耐药性的敏感度、特异度、符合率以及κ值分别为89.79%和86.54%、96.08%和94.12%、94.04%和93.38%以及0.86和0.85。对荧光PCR熔解曲线法和PCR-线性杂交酶显色法检测利福平和异烟肼耐药性结果进行比较,差异均无统计学意义(χ^(2)=0.182、0.114,P均>0.05)。结论荧光PCR熔解曲线法和PCR-线性杂交酶显色法在检测利福平和异烟肼耐药方面具有同等的诊断效能,与表型药敏试验一致性较好,且弥补了表型药敏试验检测低度耐药菌株突变的局限性,可以用于结核病的早期快速诊断,为临床及时准确进行抗结核治疗提供了依据。
Objective To analyze the drug resistance of Mycobacterium tuberculosis(MTB)infected patients in Nanjing,and evaluate the clinical value of fluorescence PCR melting curve method and PCR linear hybridization enzyme chromogenesis assay in detecting the resistance of Mycobacterium tuberculosis to rifampicin(RFP)and isoniazid(INH).Methods A total of 151 clinical isolates of Mycobacterium tuberculosis were collected in the Nanjing Public Health Medical Center.The drug resistance of MTB to RFP and INH were detected by absolute concentration drug sensitivity test,fluorescence PCR melting curve method and PCR linear hybridization enzyme chromogenesis assay.The diagnostic efficacy of the above two molecular biology methods was evaluated using the results of phenotypic susceptibility test.Results Among the 151 clinical isolates of MTB,phenotypic resistance results showed that 6 were RFP single drug resistant,9 were INH single drug resistant,and 43 were multidrug resistant.Using absolute concentration drug sensitivity test as standard,the sensitivity,specificity,coincidence rate andκvalue of MTB to RFP and INH resistance detected by fluorescence PCR melting curve method were 100.00%and 94.23%,96.08%and 96.97%,97.35%and 96.02%,0.94 and 0.91,respectively.Using absolute concentration drug sensitivity test as standard,the sensitivity,specificity,coincidence rate andκvalue of MTB to RFP and INH resistance detected by PCR linear hybridization enzyme chromogenesis assay were 89.79%and 86.54%,96.08%and 94.12%,94.04%and 93.38%,0.86 and 0.85,respectively.There were no significant differences between the two methods(χ^(2)=0.182,0.114;all P>0.05).Conclusions The fluorescent PCR melting curve method and the PCR⁃linear hybridization enzyme colorimetric method had the same diagnostic efficacy in detecting rifampicin and isoniazid resistance,had good consistency with the phenotypic drug sensitivity test,and could make up for the limitations of the phenotypic drug sensitivity test in detecting mutations of low⁃resistance strains.They could be used for early and rapid diagnosis of tuberculosis and provide a basis for timely and accurate clinical anti⁃tuberculosis treatment.
作者
胡琴琴
陈芳芳
许坦
肖园园
刘冰雪
陈影影
施旭东
李金龙
HU Qinqin;CHEN Fangfang;XU Tan;XIAO Yuanyuan;LIU Bingxue;CHEN Yingying;SHI Xudong;LI Jinlong(Department of Clinical Laboratory,Nanjing Hospital Affiliated to Nanjing University of Chinese Medicine,Nanjing,Jiangsu 210000,China;Department of Clinical Laboratory,General Hospital of Eastern Theater Command,PLA,Nanjing,Jiangsu 210000,China)
出处
《热带医学杂志》
CAS
2024年第8期1089-1093,共5页
Journal of Tropical Medicine
基金
国家自然科学基金项目(82002233)
无锡市卫生健康委面上项目(M202201)。
关键词
荧光PCR熔解曲线法
PCR-线性杂交酶显色法
结核分枝杆菌
利福平
异烟肼
Fluorescence PCR melting curve method
PCR linear hybridization enzyme chromogenesis assay
Mycobacterium tuberculosis
Rifampicin
Isoniazid
