食品工业科技2013，Vol.34Issue(13)：147-152,6.
副溶血弧菌的磁珠捕获及检测
Bead capture and detection of Vibrio parahaemolyticus
摘要
Abstract
A novel method was developed for rapid detection of Vibrio parahaemolyticus with immunomagnetic separation followed by PCR.Whole cells of V.parahaemolyticus were used as antigen for polyclonal antibody(pAb)production in rabbit.Titers of pAb were detected by enzyme-linked immunosorbent assay(ELISA).Antiserum was purified with ammonium sulfate precipitation and reverse adsorption,purity and specificity of the resulted pAb was tested with SDS-PAGE and dot-blot hybridization.Purified pAb was conjugated with magnetic beads for bacteria separation.Results showed that the titer of purified antibody was 106,and the capture rates of immunomagnetic beads were between 55％～70％ under different conditions.Salt concentration and pH had only slight influence on the capture rate.The limit of detection(LOD) of the method was 102cfu/mL in pure culture and 103cfu/mL in fish paste samples (in presence of other kinds of bacteria species).In conclusion,combined immunomagnetic separation with PCR for the detection of V.parahaemolyticus has high sensitivity and specificity,which is promising for in-field detection.关键词
副溶血弧菌/多克隆抗体/免疫磁珠/PCRKey words
Vibrio parahaemolyticus/ polyclonal antibodies/ immunomagnetic beads/ PCR分类
轻工纺织引用本文复制引用
王报贵,武晓丽,董素琴,陈飞,陈星星,甘敏,明星,徐锋..副溶血弧菌的磁珠捕获及检测[J].食品工业科技,2013,34(13):147-152,6.基金项目
"十二五"国家科技支撑计划(2011BAK10B06,2011BAK10B01,2011BAK10B02) （2011BAK10B06,2011BAK10B01,2011BAK10B02）
国家自然科学基金项目(3100008,31170091) （3100008,31170091）
食品科学与技术国家重点实验室目标导向课题(SKLF-MB-201002)、自由探索课题(SKLF-TS-200916) （SKLF-MB-201002）
2012年赣鄱英才555工程人才计划. （）
