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利用液滴数字 PCR 检测弱毒活疫苗中鸡传染性贫血病毒污染的新策略。

A New Strategy for the Detection of Chicken Infectious Anemia Virus Contamination in Attenuated Live Vaccine by Droplet Digital PCR.

机构信息

College of Veterinary Medicine, Shandong Agricultural University, Tai'an, Shandong, China.

Shandong Provincial Key Laboratory of Animal Biotechnology and Disease Control and Prevention, Tai'an, Shandong, China.

出版信息

Biomed Res Int. 2019 May 16;2019:2750472. doi: 10.1155/2019/2750472. eCollection 2019.

Abstract

Chicken infectious anemia virus () causes the atrophy of bone marrow hematopoietic and lymphoid tissues in chicks, leading to huge economic losses all over the world. The using of attenuated vaccine contaminated with CIAV increased the mortality and the pathogenicity of other diseases in many farms. However, it is difficult to detect the CIAV contamination by general detection technology due to the extremely low dose of CIAV in vaccines. In this study, we established a new method called droplet digital Polymerase Chain Reaction () to detect CIAV contamination of vaccines more sensitively and accurately. The lowest detection limitation of this method is 2.4 copies of CIAV plasmid or CIAV contamination at 0.1 EID/1000 feathers in vaccines without any positive signals of other viruses. Besides, the sensitivity of ddPCR is 100 times greater than that of conventional PCR and 10 times greater than that of real-time PCR. The ddPCR technique is more sensitive and more intuitive. Therefore, it could be valuable for the detection of CIAV contamination in vaccines.

摘要

鸡传染性贫血病毒(CIAV)可引起雏鸡骨髓造血和淋巴组织萎缩,给全世界造成巨大经济损失。使用污染有 CIAV 的减毒疫苗增加了许多农场中其他疾病的死亡率和致病性。然而,由于疫苗中 CIAV 的剂量极低,一般检测技术很难检测到 CIAV 的污染。在这项研究中,我们建立了一种称为液滴数字聚合酶链反应(ddPCR)的新方法,以更灵敏、更准确地检测疫苗中的 CIAV 污染。该方法的最低检测限为 2.4 拷贝 CIAV 质粒或疫苗中 CIAV 污染 0.1 EID/1000 羽,且没有其他病毒的任何阳性信号。此外,ddPCR 的灵敏度比常规 PCR 高 100 倍,比实时 PCR 高 10 倍。ddPCR 技术更灵敏、更直观。因此,它可能对检测疫苗中的 CIAV 污染具有重要价值。

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