摘要
目的:以金标准试剂盒为参照,对新研制的猴B病毒抗体ELISA检测试剂盒进行准确性评价,以期为我国实验灵长类动物产业提供优质廉价的检测试剂。方法:利用3个批次共表达猴B病毒gD和g B蛋白的细胞上清分别制备ELISA检测试剂盒,与金标准试剂盒(VRL的ELISA试剂盒)同时对667只食蟹猴血清进行检测,考察新研制剂盒的准确性;然后,挑选强阳性、弱阳性、阴性样品,用另外2个批次ELISA试剂盒检测,考察不同批次试剂盒检测结果的稳定性。结果:金标准试剂盒对667只食蟹猴血清抗体检测结果为阳性280份、阴性387份,而新研制的ELISA试剂盒结果为阳性282份、阴性385份;与金标准相比,阳性样品的符合率为98.93%(277/280),阴性样品的符合率为98.97%(383/387),总体符合率为98.95%(660/667)。对100份强阳性样品、70份弱阳性样品、108份阴性样品及7份差异样品的重复检测表明,除1份弱阳性(金标准为阴性)检测为阴性外,其余样品与前期一致。结论:与金标准相比,新研制的试剂盒具有较高的准确性,可用于猴B病毒抗体的检测。
Objective:In order to provide a kind of high quality and low-cost detection reagents for experimental primates industry,the accuracy of the newly developed B virus antibody ELISA test kits was evaluated withthe gold standard kit as the reference.Methods:Three batches of cell supernatants co-expressing monkey B virusgD and gB protein were used to prepare ELISA kits for B virus antibodies test.Then,667cynomolgus monkeyssera were detected simultaneously by gold standard kits(VRL ELISA kits)and new kits to investigate its accuracy.Finally,strong positive,weak positive and negative samples were selected and tested by the other two batchesof ELISA kits to examine the stability in different kits.Results:In667cynomolgus monkey sera samples,positive samples and negative samples were280and385respectively,detected in gold standard kits.The corresponding results were282and387in new ELISA kits.In positive samples,negative samples and all the samples,the coincidence rate was98.93%(277/280),98.97%(383/387)and98.95%(660/667),respectively,compared with gold standard kits.Then,100strong positive samples,70weakly positive samples,108negative samples and7differentialsamples,according to the OD values,were screened and tested with the other two batches of ELISA kits.Exceptfor one weakly positive sample(negative in gold standard kits)tuned into negative,other samples were consistentwith the previously tested.Conclusion:Compared with the gold standard ELISA kits,the newly developed kitshave high accuracy,could be used for the detection of monkey B virus antibody.
作者
叶华虎
陈建宇
周小军
王进
代艳艳
法云智
袁菊芳
YE Hua-Hu;CHEN Jian-Yu;ZHOU Xiao-Jun;WANG Jin;DAI Yan-Yan;FA Yun-Zhi;YUAN Ju-Fang(Laboratory Animal Center, Academy of Military Medical Sciences, Beijing 100071,China;College of Life Sciences, Inner Mongolia University, Hohhot 010021, China)
出处
《生物技术通讯》
CAS
2017年第5期651-656,共6页
Letters in Biotechnology
基金
国家支撑计划(2015BAI08B03)
国家自然科学基金(31272514
31501908)
