巴尔通体实时荧光定量PCR检测方法的建立  

Establishment and application of real-time fluorescence quantitative PCR assay for detection of Bartonella

作　　者：姚美琳[1] 罗炜[1] 李国伟[1] 苏丽琼[1] 叶曦[1] 

机构地区：[1]厦门市海沧区疾病预防控制中心,福建厦门361026

出　　处：《中国热带医学》2009年第10期1963-1965,共3页China Tropical Medicine

摘　　要：目的建立灵敏、特异的SYBR-Green荧光定量PCR检测巴尔通体的方法,以用于巴尔通体的实验室快速诊断和自然疫源地的流行病学研究。方法利用离心柱法抽提鼠血中巴尔通体细菌基因组DNA,在荧光定量PCR检测仪上检测基因组中枸橼酸合酶gltA的Ct含量,结合熔解曲线的Tm值进行分析,建立实时荧光定量检测法。结果检测62份阳性菌株,Ct值小于30,Tm值在81-83℃范围内;检测309份鼠血抽提的基因组DNA样品,阳性128份,阳性率为38.19%。结论建立了检测巴尔通体枸橼酸合酶gltA的SYBR-Green荧光定量PCR方法,较常规培养法更快捷、简便、敏感,适合大面积调查巴尔通体在鼠群中的分布情况,为流行病学的深入研究提供科学依据,并可作为检测巴尔通体的技术储备。Objective To develop a sensitive and specific quantitative SYBR-Green fluorescent PCR assay for rapid diagnosis of Bartonella and its natural foci. Methods Bartonella genomic DNA was extracted from rat blood by means of centrifuge column method. Real-time fluorescent quantitative assay was developed based on analysis of the Ct content of citrate synthase ghA combined with the value of Tm of melting curve by using the fluorescence quantitative PCR detection detector. Results There 62 positive strains were detected,Ct value was less than 30 and Tm values in the range of 81-83 ℃; 309 genomic DNA were extracted from rat blood, 128 were positive with a positive rate of 38.19%. Conclusion Realtime PCR are faster,more convenient,and sensitive for diagnosis of Bartonella infections,comparod with traditional culture. This methods may be suitable for large-area survey of the distributioin of BartoneUa in rat population, which will be usful for the epidemiologieal in-depth study of Bartonella.

关 键 词：巴尔通体 SYBR—Green 荧光定量PCR 

分 类 号：R446.6[医药卫生—诊断学]