BTV核酸检测能力验证样品制备及其应用评价

发布时间：2019-01-04 19:37

【摘要】：为了解国内蓝舌病病毒(bluetongue virus,BTV)核酸检测的整体水平,督促实验室保持和提高BTV核酸检测的能力,设计和实施了"CNCA-15-A02《蓝舌病病毒核酸检测》能力验证计划"。本次能力验证制备了BTV阴性,强阳性和弱阳性样品。无菌采集健康山羊抗凝血加少量TRIzol后制备阴性样品;将BTV1型细胞培养物通过TRIzol处理灭活病毒后,用阴性样品做适当稀释制备了BTV强阳性和弱阳性样品;均匀性和稳定性检验证实样品均达到中国合格评定国家认可委员会对能力验证样品的要求。将能力验证样品编号后通过快递的方式下发给参试实验室,共有21个省、市的31实验室参加了本次能力验证活动,其中1家实验室选用了套式RT-PCR进行检测,其余30家均选用荧光RT-PCR方法,报送结果经比对验证均为满意,满意率达100%。本次能力验证对提升我国BTV的检测水平和评估各级实验室检测能力具有重要意义,为我国蓝舌病的诊断和防控工作提供技术保障。
[Abstract]:In order to understand the whole level of nucleic acid detection of Bluetongue virus (bluetongue virus,BTV) in China and urge the laboratory to maintain and improve the ability of BTV nucleic acid detection, a "CNCA-15-A02" blue-tongue virus nucleic acid detection "capability verification program was designed and implemented. BTV negative, strong positive and weak positive samples were prepared. After anticoagulant and a small amount of TRIzol were collected aseptically, BTV1 cell culture was treated with TRIzol to inactivate the virus, and the strong and weak positive BTV samples were prepared by dilution of the negative samples. The homogeneity and stability test confirmed that the samples met the requirements of the China National Accreditation Committee. The capacity verification sample number was sent to the test laboratory by express delivery. A total of 31 laboratories from 21 provinces and cities participated in the capability verification activity. Among them, one laboratory selected nested RT-PCR for testing. Fluorescence RT-PCR method was used in the other 30 families. The results were verified by comparison and the satisfaction rate was 100. This capability verification is of great significance to improve the detection level of BTV in China and to evaluate the detection ability of laboratories at all levels, and provide technical guarantee for the diagnosis, prevention and control of bluetongue in China.
【作者单位】：北京出入境检验检疫局;云南出入境检验检疫局;
【基金】：国家质检总局科研项目(2014IK249)
【分类号】：S852.65

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