摘要
目的建立巴贝西虫数字PCR检测的有效方法,为后续巴贝西虫研究及我国血供安全评估提供技术方法储备和参考数据。方法从GenBank下载我国可感染人巴贝西虫的18S rRNA基因序列,并根据序列共同部分设计引物,建立基于巴贝西虫灵敏度高、可绝对定量的数字PCR检测方法;检测1000(人)份2016年7月19日~2016年8月24日牡丹江市采集的献血者红细胞标本。结果所建立的巴贝西虫数字PCR检测方法重复性良好,可稳定检测单拷贝数核算,应用于献血者标本数字PCR阳性检出率0.2%(2/1000),2例巴贝西虫感染者经间接免疫荧光检测(IFA)结果为阳性,初步判断为无症状感染。结论巴贝西虫数字PCR检测方法具有较高灵敏度和较好稳定性;若能应用于献血者巴贝西虫血液筛查,可减少这类无症状感染对血液安全的威胁。
Objective To establish an efficient digital PCR method for Babesia detection,so as to provide data reference for the follow-up studies and the evaluation of blood supply safety in China.Methods 18S rRNA conservative gene sequence of Babesia spp.was downloaded from Genbank,and primers were designed according to the common part of the sequence to establish a highly sensitive and absolutely quantitative digital PCR detection method for Babesia detection.A total of 1000 red blood cell samples collected from voluntary blood donors in Mudanjiang City from July 19,2016 to August 24,2016 were detected using this new digital PCR method.Results The established digital PCR method for Babesia detection showed a good repeatability and could steadily detect the mono-copied nuclear acid.The positive rate was 0.2%(2/1000)by this method.The two blood donors were further confirmed positive by Indirect Immunofluorescence Assay(IFA),and preliminarily judged as asymptomatic infection.Discussion The digital PCR method for Babesia detection established in this study has high sensitivity and stability,and the its application on blood screening is conductive to reduce the threats of asymptomatic infection to blood safety.
作者
杨亚闪
林宝钗
刘曹毅
高阳
何苗
刘鱼
YANG Yashan;LIN Baochai;LIU Caoyi;GAO Yang;HE Miao;LIU Yu(Institute of Blood Transfusion,Chinese Academy of Medical Sciences&Peking Union Medical College,Chengdu 610052,China)
出处
《中国输血杂志》
CAS
2021年第8期835-839,共5页
Chinese Journal of Blood Transfusion
基金
四川省科学技术厅重点研发项目(2019YFS0319)。
