空肠弯曲菌免疫磁珠富集方法的建立  

The Method for Enrichment of Campylobacter Jejuni Based on Immunomagnetic Beads

作　　者：王壮[1] 欧瑜[1] 唐泰山[2] 祝长青[2] 栾军[2] 蒋原[2] 张常印[2] 

机构地区：[1]中国药科大学生命科学与技术学院,江苏南京210009 [2]江苏出入境检验检疫局动物检疫实验室,江苏南京210001

出　　处：《药物生物技术》2014年第1期43-47,共5页Pharmaceutical Biotechnology

基　　金：国家质检总局科技项目(No.2008IK145)

摘　　要：为建立空肠弯曲菌富集方法,研究利用空肠弯曲菌特异性多表位人工多肽的特异性抗体包被磁珠,制成免疫磁珠富集空肠弯曲菌。在不同稀释梯度的菌液中加入免疫磁珠,富集目标菌,铺于CFA显色平板培养,统计菌落个数,确定免疫磁珠的敏感性;在非空肠弯曲菌和弯曲菌中分别添加空肠弯曲菌,用免疫磁珠富集,确定免疫磁珠的特异性。结果表明2 mg免疫磁珠可以在1 mL空肠弯曲菌菌液（浓度为2.5×103/mL）中稳定富集到空肠弯曲菌,统计菌落数为100~300 CFU,具有较好的敏感性;在干扰菌中富集空肠弯曲菌,铺板培养后,荧光PCR鉴定为阳性,富集不受干扰菌影响。用建立的免疫磁珠富集方法富集采集样品中的空肠弯曲菌,结果显示该方法有较好的特异性,具有较好的应用前景。To establish the method of detection of Campylobacter jejuni,the specific antibody of Campylobacter jejuni to coat the magnetic beads was used to enrich Campylobacter jejuni.To determine the sensitivity of the immunomagnetic beads,it was used to enrich the Campylobacter jejuni in different diluted solutions.Then the bacteria enriched by the immunomagnetic beads in the CFA medium was cultured and the colonies was statisticed.To determine the specificity of the immunomagnetic beads,it was used to enrich the Campylobacter jejuni which added in the non-Campylobacter jejuni and other Campylobacter respectively.The results indicate that 2 mg immunomagnetic beads can enrich the target bacteria in the solution of which the dilution was 2.5 × 103/mL.The number of the colonies of the bacteria was 100 ~ 300 CFU.The immunomagnetic beads enriched Campylobacter jejuni added in different interference strains which was cultured in the CFA medium.The suspected colony was detected by real-time PCR.The results were positive.The Campylobacter jejuni in the samples was detected by this method successfully.This method has a good prospects.

关 键 词：空肠弯曲菌 特异性抗体 免疫磁珠 富集 添加 干扰菌 

分 类 号：R737.9[医药卫生—肿瘤]