摘要
目的:采用细菌基因外重复回文序列扩增(REP-PCR)和肠道细菌基因间重复序列扩增(ERIC-PCR)两种方法对副溶血性弧菌2株标准株,13株实验室保存菌株和73株分离菌株共88株进行分子分型。方法:通过REP和ERIC-PCR指纹图谱扩增,利用NTsys-pc软件采用Dice系数对指纹图谱进行聚类结果分析。结果:所有供试菌株可通过此两种方法进行分型得到清晰的指纹图谱,并反映出副溶血性弧菌具有丰富的遗传多样性,REP-PCR扩增出5~9条分布在609~4200bp之间的条带,可将副溶血性弧菌分为5个群12类型,分辨指数达0.93,其中tdh+株在相似系数0.86时可聚类在第Ⅰ群;ERIC-PCR扩增出5~11条分布在400~7593bp之间的条带,将副溶血性弧菌分为7个群11个类型,分辨指数为0.94,其中tdh+株在相似系数0.76时可聚类在第Ⅰ群。结论:两种方法均显现出很好的分型能力,能够很好地将环境分离tdh+菌株和标准菌株聚类在一起,其中REP-PCR较ERIC-PCR重复性更好。
Abstract: Objective: The enterobacterial repetitive intergenic consensus sequence-polymerase chain reaction (ERIC-PCR) and repetitive extragenic palindromic element-polymerase chain reaction (REP-PCR) were used to analyze the types of 88 l, Tbrio parahaemolyticus including 2 standard strains, 13 laboratory strains and 73 isolated strains. Methods: REP and ERIC primers were used to amplify the repeated sequences. NTsys-pc soft-ware was used to calculate the discriminative index of REP- and ERIC-PCR with Dice coefficients. Results: All 88 tested V. parahamolyticus could be typed by REP- and ERIC- PCR clearly and revealed abundant genetic diversity in V. parahamolyticus. According to the results of REP-PCR-amplified 5-9 bands within 609-4200 bp, E parahamolyticus was typed into 5 groups and 12 types with discrimination index of 0.93, and the tdh~ strains with resolving index of 0.86 could be classified in Group I. According to ERIC-PCR-amplified 5-11 bands within 400-7593 bp, V. parahamolyticus was typed into 7 groups and 11 types with discrimination index of 0.94, and the tdh strains with discrimination index of 0.76 could be classified in group I. Conclusion: The isolated tdh strains are clustered together with standard strains using the two methods. Although both methods exhibit good typing capability, REP- PCR is more repeatable and stable than ERIC-PCR.
出处
《食品科学》
EI
CAS
CSCD
北大核心
2013年第10期263-267,共5页
Food Science
基金
霍英东教育基金会第十一届高等院校优选课题(114035)
上海市科学技术委员会部分地方院校能力建设项目(11310501100)
上海市教育委员会重点学科建设项目(J50704)
关键词
副溶血性弧菌
TDH
肠内细菌基因组间重复序列分析
细菌基因外重复回文序列扩增
分型
Vibrio parahaemolyticus: tdh enterobacterial repetitive intergenic consensus sequence-polymerase chainreaction (ERIC-PCR)
repetitive extragenic palindromic-polymerase chain reaction (REP-PCR)
typingg