摘要
目的 应用TaqMan-MGB探针实时荧光定量PCR技术快速检测青海省海西州鼠疫自然疫源地鼠疫菌耐链霉素基因,为今后该地区突发人间鼠疫的精准临床用药提供理论依据。方法 分离培养海西地区1957—2009年间取自鼠疫患者、媒介昆虫及中间宿主的代表性鼠疫菌110株,提取其DNA,针对我国链霉素耐药基因rpsl基因设计引物P-F和P-R和TaqMan-MGB探针Probe1 [FAM]和Probe2[VIC],利用荧光定量PCR技术,进行耐药rpsl基因筛查。结果 110株被试菌株中FAM检测均为阳性(RFU峰值>2000);VIC阳性的为0株(RFU峰值<200)。阳性对照和空白对照成立。结论 实时荧光定量PCR结果显示,该地区未检测出耐链霉素菌株。
Objective To rapidly detect streptomycin resistance genes of Yersinia pestis in the Natural Focus of Plague of Haixi prefecture in Qinghai province by TaqMan MGB probe fluorescence real-time quantitative PCR,and to provide the theoretical basis for precise clinical drug use of plague emergencies in this area.Methods 110 representative strains of Yersinia pestis collected from 1957 to 2009 from plague patients,vector insects and intermediate hosts were isolated and cultured,with their DNA being extracted.Probe1[FAM]and Probe2[VIC],which were probes of Primers P-F and P-R and TaqMan-MGB,were designed for the rpsl gene,which was a streptomycin resistance gene often found in China,and fluorescence quantitative PCR was utilized to screen the rpsl gene of plague resistance genes.Results In FAM detection,all 110 strains had positive value(RFU peak>2000).No strains were positive in VIC detection(RFU peak<200).The positive control and the blank control proved to be effective.Conclusion Results of real-time quantitative PCR demonstrated that streptomycin resistant strains were not detected in the tested strains in this area.
作者
张琪
李胜
靳娟
何建
杨晓艳
辛有全
柏吉祥
周奎章
张晓璐
蒋可
代瑞霞
Zhang Qi;Li Sheng;Jin Juan;He Jian;Yang Xiao-yan;Xin You-quan;Bai Ji-xiang;Zhou Kui-zhang;Zhang Xiao-u;Jiang Ke;Dai Rui-xia(Specialized Laboratory of Yersinia pestis,Qinghai Institute for Endemic Disease Prevention and Control,Xi'ning 810021;Department of Plague,Qinghai Institute for Endemic Disease Prevention and Control,Xi'ning 810021;Department of Public Health,Medical School,Qinghai University,Xi'ning 810001)
出处
《中国抗生素杂志》
CAS
CSCD
北大核心
2023年第10期1198-1200,I0001,共4页
Chinese Journal of Antibiotics
基金
国家自然科学基金项目(No.82260401)。
