In the present study, a nanometer PCR (nano-PCR) assay was developed to detect the Africa swine fever virus (ASFV) using the P54 gene containing recombinant plasmid as the tamplate for 552bp fragment amplification. The amplification was efficiently enhanced with the gold nanoparticles as thermal mediator in the amplification system, and the sensitivity of the nano-PCR was more than 1,000 times than that of conventional PCR with a detection limit of 10 copies and had no cross-reactions with E. coli, porcine pseudorabies virus, porcine circovirus type Ⅱ, classical swine fever virus, porcine reproductive and respiratory syndrome virus, porcine Teschovirus 8 and encephalomyocarditis virus. Furthermore, a total of 69 clinical samples from Heilongjiang, Jilin and Henan provinces were detected by this assay. The results showed that all the samples were negative, indicating no ASFV infection occurred in these areas.%为将纳米PCR技术引入非洲猪瘟病毒(ASFV)检测,本研究建立了ASFV的高效纳米PCR检测方法,该方法采用纳米金颗粒作为热导介子,提高了PCR反应效率,采用该方法同时检测ASFV、大肠杆菌、猪伪狂犬病毒、猪圆环病毒2型、典型猪瘟病毒、猪繁殖与呼吸综合征病毒、猪捷申病毒、猪脑心肌炎病毒等,结果只有ASFV能够扩增出552 bp的目的片段.敏感性试验表明纳米PCR检测技术是常规PCR检测技术敏感性的1 000倍以上,最低核酸拷贝数检出量可以达到10个拷贝.采用该方法对黑龙江、吉林和河南3个省份的8个地区临床送检的69份样品进行检测,检测结果显示均为阴性,在以上3个地区均未发现ASFV感染情况.
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