用于非洲猪瘟病毒检测与研究的液滴数字聚合酶链反应（ddPCR）的开发与应用
Development and application of a droplet digital polymerase chain reaction (ddPCR) for detection and investigation of African swine fever virus.
作者信息
Wu Xulong, Xiao Lu, Lin Hua, Chen Shijie, Yang Miao, An Wei, Wang Yin, Yang Zexiao, Yao Xueping, Tang Zizhong
机构信息
College of Veterinary Medicine, Sichuan Agricultural University, Chengdu 611130, China (Xulong Wu, Yin Wang, Zexiao Yang, Xueping Yao); Sichuan Animal Science Academy, Chengdu, 610066, China (Lu Xiao); Key Laboratory of Animal Genetics and Breeding of Sichuan Province, Chengdu, 611130, China (Lu Xiao); Key Laboratory of Animal Disease and Human Health of Sichuan Province, Chengdu 611130, China (Yin Wang, Zexiao Yang, Xueping Yao); Inspection and Quarantine Technical Center, Sichuan entry-exit inspection and Quarantine Bureau, Chengdu 610041, China (Hua Lin, Shijie Chen, Miao Yang, Wei An); College of Life Science, Sichuan Agricultural University, Yaan 625014, China (Zizhong Tang).
出版信息
Can J Vet Res. 2018 Jan;82(1):70-74.
The aim of this study was to develop a droplet digital polymerase chain reaction (ddPCR) method to detect African swine fever virus (ASFV). The methods of ASFV real-time PCR and ddPCR were established and optimal reaction conditions were confirmed. Each method was evaluated for linearity, limit of detection, and specificity. The results indicated that ASFV ddPCR had a high degree of linearity ( ≥ 0.998) and specificity. The detection limit was 10 copies/reaction, which was approximately a 10-fold greater sensitivity than real-time PCR. This sensitive method could be used as an efficient molecular biology tool to diagnose ASFV, which is very important for preventing the spread of diseases across borders.
本研究的目的是开发一种用于检测非洲猪瘟病毒（ASFV）的液滴数字聚合酶链反应（ddPCR）方法。建立了ASFV实时PCR和ddPCR方法，并确定了最佳反应条件。对每种方法的线性、检测限和特异性进行了评估。结果表明，ASFV ddPCR具有高度的线性（≥0.998）和特异性。检测限为10拷贝/反应，其灵敏度比实时PCR高约10倍。这种灵敏的方法可作为诊断ASFV的有效分子生物学工具，这对于防止疾病跨境传播非常重要。
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