中国动物检疫
中國動物檢疫
중국동물검역
CHINA ANMAL QUARANTINE
2015年
4期
79-83
,共5页
赵丽%蔡阳%黄伟%邱杨%刘建丽%陈进会%王振国
趙麗%蔡暘%黃偉%邱楊%劉建麗%陳進會%王振國
조려%채양%황위%구양%류건려%진진회%왕진국
空肠弯曲菌%双抗体夹心免疫学检测%液相芯片检测
空腸彎麯菌%雙抗體夾心免疫學檢測%液相芯片檢測
공장만곡균%쌍항체협심면역학검측%액상심편검측
Camplobacter jejuni%double antibody sandwich immunoassay%liquid phase chip detection
[目的]建立空肠弯曲菌液相芯片检测方法。[方法]本研究将空肠弯曲菌HIP蛋白单克隆抗体与聚苯乙烯微球偶联,结合双抗体夹心技术建立空肠弯曲菌液相芯片检测方法,测定不同浓度的抗体与微球偶联率。通过L9(34)正交设计试验优化方法的反应条件,并进行灵敏度和特异性试验。[结果]较优实验条件即多克隆抗体工作浓度为1:100、生物素标记的羊抗兔IgG工作浓度为1:500、SA-PE的工作浓度为10ug/mL、生物素标记的抗体与SA-PE反应时间为90min。该方法灵敏度可达103CFU/mL,与其他常见食源性致病菌无交叉反应。[结论]该方法灵敏度高、特异性强、重复性好,可快速检测食品中的空肠弯曲菌。
[目的]建立空腸彎麯菌液相芯片檢測方法。[方法]本研究將空腸彎麯菌HIP蛋白單剋隆抗體與聚苯乙烯微毬偶聯,結閤雙抗體夾心技術建立空腸彎麯菌液相芯片檢測方法,測定不同濃度的抗體與微毬偶聯率。通過L9(34)正交設計試驗優化方法的反應條件,併進行靈敏度和特異性試驗。[結果]較優實驗條件即多剋隆抗體工作濃度為1:100、生物素標記的羊抗兔IgG工作濃度為1:500、SA-PE的工作濃度為10ug/mL、生物素標記的抗體與SA-PE反應時間為90min。該方法靈敏度可達103CFU/mL,與其他常見食源性緻病菌無交扠反應。[結論]該方法靈敏度高、特異性彊、重複性好,可快速檢測食品中的空腸彎麯菌。
[목적]건립공장만곡균액상심편검측방법。[방법]본연구장공장만곡균HIP단백단극륭항체여취분을희미구우련,결합쌍항체협심기술건립공장만곡균액상심편검측방법,측정불동농도적항체여미구우련솔。통과L9(34)정교설계시험우화방법적반응조건,병진행령민도화특이성시험。[결과]교우실험조건즉다극륭항체공작농도위1:100、생물소표기적양항토IgG공작농도위1:500、SA-PE적공작농도위10ug/mL、생물소표기적항체여SA-PE반응시간위90min。해방법령민도가체103CFU/mL,여기타상견식원성치병균무교차반응。[결론]해방법령민도고、특이성강、중복성호,가쾌속검측식품중적공장만곡균。
Objective]The aim of the study is to establish liquid chip method for detection of Camplobacter jejuni. [Method] HIP protein monoclonal antibody againstCamplobacter jejuni was coupled with polystyrene microspheres and a liquid chip method was established for detection ofCamplobacter jejuni in combination with double antibody sandwich technique. The coupling rate of different antibody concentration with the microsphere was determined.The reaction conditions were optimized through L9(34)orthogonal design test,and the developed method was evaluated for its sensitivity and specificity. [Result] The results showed that the optimal test conditions were:the working poly-antibody concentration was 1:100,the working concentration of biotin labeled goat anti rabbit IgG was 1:500,the working concentration of SA-PE was10ug/mL and the reaction time of biotin labeled antibody and SA-PE was 90min. The sensitivity of the assay reached 103CFU/mL,without cross reaction with other common food borne pathogenic bacteria. [Conclusion]The method was of high sensitivity,specificity and reproducibility and could be used for rapid detection ofCamplobacter jejuni in food.