通过基于PCR的快速特异性检测方法检测大肠杆菌2型多糖荚膜合成基因kpsM
Detection of the Escherichia coli group 2 polysaccharide capsule synthesis Gene kpsM by a rapid and specific PCR-based assay.
作者信息
Johnson James R, O'Bryan Timothy T
机构信息
Mucosal and Vaccine Research Center, Minneapolis VA Medical Center, Minnesota.
出版信息
J Clin Microbiol. 2004 Apr;42(4):1773-6. doi: 10.1128/JCM.42.4.1773-1776.2004.
Abstract
A rapid and simple PCR-based assay for detection of the group 2 capsule synthesis gene kpsM of Escherichia coli was designed and validated. When combined with the published group 2 primers (kpsIIf, 5'-GCGCATTTGCTGATACTGTTG-3'; kpsIIr, 5'-CATCCAGACGATAAGCATGAGCA-3'), the new primers (the kpsIIf primer and a new reverse primer K2r, 5'-AGGTAGTTCAGACTCACACCT-3') allowed specific identification by exclusion of the heretofore elusive K2 kpsM variant. The primers yielded the predicted amplicon when multiplexed with other primers and used under varied assay conditions, including a range of concentrations of individual reaction mixture ingredients and of annealing temperatures (from 54 to 64 degrees C).
摘要
设计并验证了一种基于聚合酶链反应（PCR）的快速简便检测方法，用于检测大肠杆菌2型荚膜合成基因kpsM。当与已发表的2型引物（kpsIIf，5'-GCGCATTTGCTGATACTGTTG-3'；kpsIIr，5'-CATCCAGACGATAAGCATGAGCA-3'）结合使用时，新引物（kpsIIf引物和新的反向引物K2r，5'-AGGTAGTTCAGACTCACACCT-3'）通过排除迄今难以捉摸的K2 kpsM变体实现特异性鉴定。当与其他引物进行多重扩增并在不同的检测条件下使用时，包括各种浓度的单个反应混合物成分和退火温度（54至64摄氏度），这些引物产生了预期的扩增产物。
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