Shangkuan Y H, Lin H C, Wang T M
Division of Bacteriology, Institute of Preventive Medicine, National Defense Medical Center, Sanhsia, Taipei, Taiwan, ROC.
J Appl Microbiol. 1997 Mar;82(3):335-44. doi: 10.1046/j.1365-2672.1997.00365.x.
Vibrio cholerae strains isolated from patient, food and environmental sources in Taiwan and reference V. cholerae strains were examined by repetitive element sequence-based PCR (rep-PCR). Specimens from broth cultures were used directly in the PCR mixture with three different primers. The PCR fingerprinting profiles of toxigenic 01 isolates were not only homogeneous with primers from enterobacterial repetitive intergenic consensus (ERIC) sequences, but also allowed the differentiation from non-toxigenic O1 and non-O1 strains. Toxigenic 01 strains were further differentiated into El Tor and classical biotypes with primers designed from ERIC-related sequences of V. cholerae. Primers from the other V. cholerae repetitive DNA sequences, VCR, separated toxigenic El Tor strains into six groups and a unique pattern was also obtained in 16 isolates from imported cases of cholera and imported seafood. The results indicated that rep-PCR can be used to identify and differentiate different toxigenic 01, non-toxigenic 01 and non-O1 V. cholerae isolates.
采用基于重复元件序列的聚合酶链反应(rep-PCR)对从台湾患者、食物及环境来源分离出的霍乱弧菌菌株以及霍乱弧菌参考菌株进行检测。肉汤培养物的标本直接用于含有三种不同引物的聚合酶链反应混合物中。产毒O1分离株的聚合酶链反应指纹图谱不仅与来自肠杆菌重复基因间共有序列(ERIC)的引物具有同源性,还能与非产毒O1和非O1菌株区分开来。利用根据霍乱弧菌ERIC相关序列设计的引物,可将产毒O1菌株进一步分为埃尔托生物型和古典生物型。来自霍乱弧菌其他重复DNA序列VCR的引物,将产毒埃尔托菌株分为六组,并且在16株来自霍乱输入病例和进口海产品的分离株中也获得了独特的图谱。结果表明,rep-PCR可用于鉴定和区分不同的产毒O1、非产毒O1和非O1霍乱弧菌分离株。