摘要
目的:建立一种基于qRT-PCR的微量细胞中和实验改良法,快速、定量检测汉滩病毒感染中的中和抗体活性。方法:将本室前期制备的抗汉滩病毒高中和活性鼠源性单抗3G1、鼠/人嵌合单抗1D9和感染剂量为100 TCID50的病毒混合液于37℃孵育1 h后感染Vero-E6细胞,提取细胞总RNA。根据GenBank数据库中汉滩病毒76-118株S基因序列设计特异性引物,在上述实验基础上以细胞总RNA为检测样品,建立qRT-PCR快速、定量检测汉滩病毒感染中中和抗体活性的实验方法。结果和结论:建立了一种基于qRT-PCR的微量细胞中和实验改良法,可定量测定汉滩病毒感染中的中和抗体活性,该法具有快速、灵敏、特异和重复性好等优点。
Objective： To develop a fast and accurate evaluation system for neutralization antibodies against Han- taan virus. Methods： Vero-E6 cells were incubated at 37℃ for 1 hour in the presence of Hantaan virus at 100 TCID50 infectious dose and candidate antibody（mAb 3G1 or murine/human chimeric mAb 1D9）. The total cellular RNAs from the incubation mix were extracted. The specific primers were designed according to the S gene sequence of Hantavirus 76-118 in the GenBank database. The extracted total RNAs as template, qRT-PCR was performed to detect the effect of neutralization antibodies for Hantaan virus. Results ＆ Conclusion： The method called qRT-PCR-based micro-cell-neutralization test was successfully established, which is capable to quantify the neutralizing antibodies rapidly and aeurrately.
出处
《生物技术通讯》
CAS
2017年第4期524-527,共4页
Letters in Biotechnology
基金
国家自然科学基金(31470890
31270978)
陕西省科技统筹创新工程计划(2013KTCL03-06)
军队重点课题(BWS13G024)
关键词
汉滩病毒
QRT-PCR
中和抗体
微量中和实验
肾综合征出血热
Hantaan virus
qRT-PCR
neutralization antibody
micro-neutralization test
hemorrhagic fever with renal syndrome
