聚合酶链反应检测白喉杆菌的实验研究  

An experimental study on detection of corynebacterium diphtheria by polymerase chain reaction

作　　者：王晓春[1] 熊鸿燕[1] 张建中[2] 何利华[2] 尹焱[2] 

机构地区：[1]第三军医大学预防医学系流行病学教研室,重庆400038 [2]中国疾病预防控制中心传染病预防控制所诊断室,北京102206

出　　处：《临床检验杂志》2004年第2期128-130,共3页Chinese Journal of Clinical Laboratory Science

摘　　要：目的　建立检测白喉杆菌的聚合酶链反应 (PCR)方法。方法　用PCR扩增白喉杆菌特异的白喉外毒素B基因 (toxB)片段 ,琼脂糖凝胶电泳检测扩增产物。结果　白喉杆菌参考菌株 (有毒株 )均扩增出 318bp的特异性片段 ,而其他需鉴别诊断的常见病原菌均未扩增出此特异条带。检测灵敏度为 85 0fg/ μl基因组DNA。 结论　依据toxB基因建立的白喉杆菌PCR检测方法具有高度的敏感性和特异性 。Objective The aim of this study was to explore and establish a rapid method for detecting Corynebacterium diphtheria by polymerase chain reaction (PCR).Methods The specific fragment of diphtheria toxin B gene (toxB) was detected by PCR.PCR amplification products were examined by agarose gel electrophoresis.The specificity and sensitivity of this method were evaluated.Results An 318 bp fragment specific to toxB gene were amplified in all the tested Corynebacterium diphtheria strains,whereas it was detected in none of the other respiratory pathogenetic bacteria tested.The lowest limit of detection was 850 fg/μl of genome DNA of Corynebacterium diphtheria strain.Conclusion These results indicated that the PCR assay based on toxB gene had high specificity and sensitivity and may be valuable for the diagnosis and differential diagnosis of diphtheria when the cultures of bacteria are not available.

关 键 词：聚合酶链反应 检测 白喉杆菌 实验研究 白喉外毒素B片段基因 

分 类 号：R446.5[医药卫生—诊断学]