戊型肝炎抗原检测试剂在供血浆人群的应用  

Application of ELISA kit for the detection of hepatitis E virus antigen in plasma donations

作　　者：黄维金[1] 宋爱京[1] 乔杉 赵晨燕[1] 贾雪荣 张燕 王佑春[1] 

机构地区：[1]中国食品药品检定研究院,卫生部生物技术产品检定方法与标准化重点实验室,北京100050 [2] 北京万泰股份有限责任公司,102200 [3] 成都蓉生药业有限责任公司,610041

出　　处：《中华微生物学和免疫学杂志》2016年第4期300-304,共5页Chinese Journal of Microbiology and Immunology

基　　金：国家自然科学基金面上项目（81371830,81171549）

摘　　要：目的：分析戊型肝炎病毒（ HEV）抗原检测试剂在供血浆人群和戊型肝炎系列阳转血清的检测情况,评价HEV抗原（ HEV-Ag）检测试剂的性能。方法从36340份样品中选择HEV-Ag检测阳性的样品,利用实时荧光HEV RNA检测试剂检测其核酸,并对核酸阳性者利用巢式RT-PCR的方法扩增检测HEV 的ORF2片段,对阳性扩增产物进行序列测定确认HEV、构建基因进化树划分基因型。同时,利用HEV-Ag试剂和HEV核酸试剂平行检测Biomex的戊型肝炎系列阳转血清,比较其相关性。结果在36340样品中有26份HEV-Ag阳性样品（0.07%）,利用巢式RT-PCR方法检测阳性比例为25/26,序列确认后的比例为23/26。在供血浆人群中HEV RNA确认阳性的比例为1∶1580（0.06%）。基因序列分型表明基因1型17份（74%）,基因4型6份（26%）。戊型肝炎系列血清检测发现HEV-Ag和HEV RNA相关性好,HEV-Ag阳性样品其HEV RNA均为阳性,与HEV RNA的滴度峰值一致。结论 HEV-Ag试剂与HEV RNA的相关性高,HEV-Ag筛查可作为降低血源戊型肝炎感染的一种策略。Objective To analyze the practicability of using ELISA kit for the detection of hepati-tis E virus antigen （ HEV-Ag） in plasma donations and Biomex HEV seroconversion panels. Methods The HEV-Ag positive samples were screened out from 36 340 donated blood plasma samples. Real-time fluores-cent PCR was performed for the detection of HEV RNA in HEV-Ag positive samples. The open reading frame 2 （ORF2） in HEV RNA was amplified by nested RT-PCR and the amplified products were confirmed by sequencing analysis. Phylogenetic tree was constructed for HEV genotyping. Five Biomex HEV serocon-version panels were used in this study for the detection of HEV-Ag, anti-HEV antibody and HEV RNA as well as the correlation analysis between HEV-Ag and HEV RNA. Results Twenty-six out of 36 340 plasma samples （0. 07%） were positive for HEV-Ag. Of the 26 samples, 25 samples were positive for HEV RNA as indicated by the results of nested RT-PCR and 23 positive samples were confirmed by sequencing analysis. The positive rate of HEV RNA in blood plasma donators was 1 ∶ 1 580 （0. 06%）. There were 17 samples of genotype 1 （74%） and 6 samples of genotype 4 （26%） according to the phylogenetic tree analysis. All of the HEV-Ag positive samples were also positive for HEV RNA as indicated by the analysis of Biomex sero-conversion panels. HEV-Ag was consistent with the peak of the HEV RNA concentration. Conclusion A close relationship between HEV-Ag and HEV RNA was observed. HEV-Ag screening could be used as a measure to reduce the risk of HEV transmission by blood transfusion.

关 键 词：戊型肝炎病毒 抗原 核糖核酸 血液 

分 类 号：R512.6[医药卫生—内科学] R440[医药卫生—临床医学]