基于全自动毛细管电泳技术建立的单核细胞增生李斯特氏菌MLVA分型方法  

Development of a multiple-locus variable number of tandem repeat typing method for Listeria monocytogenes upon the capillary electrophoresis

作　　者：李秀娟[1] 崔玲玲[2] 赵冬[1] 潘琢[1] 高伟利[1] 

机构地区：[1]石家庄市疾病预防控制中心,石家庄050011 [2]河北师范大学,石家庄050000

出　　处：《中国人兽共患病学报》2015年第9期839-844,共6页Chinese Journal of Zoonoses

摘　　要：目的建立针对食品来源的单核细胞增生李斯特氏菌(Listeria monocytogenes,Lm)分离株的多位点串联重复序列分型(Multiple-Locus Variable number tandem repeat Analysis,MLVA)方法,为暴发确认和溯源检测提供实验室支持。方法对2005—2014年间分离自食品的91株Lm进行14个可变数目串联重复序列(Variable Number of Tandem Repeats,VNTR)位点的检测,评估最优检测位点组合并分析检测结果。结果通过采用软件分析,由LMV1、LMV2、LMV7、Lm10、Lm11、Lm23、LM-TR6、TR3和Lm15等9个VNTR位点组成的位点组合为最优MLVA检测位点,可以将91株Lm分离株分为70个型别,分型能力达到0.987 1。结论本研究建立的基于全自动毛细管电泳的由9个检测位点组成的Lm的MLVA分型方法,具有操作简便、快速、结果客观、操作标准化、易于在不同实验室间比较的优势,可作为一线检测方法用于李斯特菌病的暴发确认和溯源检测。In order to develop a multiple-locus variable number tandem repeat analysis （MLVA） method for Listeria mono- cytogenes （L. monocytogenes）, a total of 14 variable number of tandem repeats （VNTR） loci were used to detect the 91 L. rnonocytogenes isolated from food. Results showed that the optimal combination of loci consisted of 9 loci （LMV1, LMV2, LMV7, Lml0, Lmll, Lm23, LM-TR6, TR3, Lm15）, which produced the high level of discriminatory ability with Simpson index of 0. 987 1 for foodborne L. monocytogenes, could divided 91 L. monocytogenes isolates into 70 subtypes. This method required only one conventional PCR followed by automated capillary electrophoresis, providing high discriminatory ability. It was simple, easy to perform, relatively fast, inexpensive, objective, standardized operating and could provide conveniently in- terlaboratory comparisons, which would be useful in outbreak investigations and listeriosis surveillance as a first line screening method.

关 键 词：单核细胞增生李斯特氏菌 多位点串联重复序列分型 毛细管电泳 

分 类 号：R378.99[医药卫生—病原生物学]