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用于检测非洲猪瘟病毒的快速灵敏重组酶聚合酶扩增与侧向流动试纸条联用技术

Rapid and Sensitive Recombinase Polymerase Amplification Combined With Lateral Flow Strip for Detecting African Swine Fever Virus.

作者信息

Miao Faming, Zhang Jingyuan, Li Nan, Chen Teng, Wang Lidong, Zhang Fei, Mi Lijuan, Zhang Jinxia, Wang Shuchao, Wang Ying, Zhou Xintao, Zhang Yanyan, Li Min, Zhang Shoufeng, Hu Rongliang

机构信息

Institute of Military Veterinary Medicine, Academy of Military Medical Science, Changchun, China.

College of Life Science, Ningxia University, Yinchuan, China.

出版信息

Front Microbiol. 2019 May 15;10:1004. doi: 10.3389/fmicb.2019.01004. eCollection 2019.

Abstract

African swine fever virus (ASFV), the etiological agent of African swine fever (ASF), a hemorrhagic fever of domestic pigs, has devastating consequences for the pig farming industry. More than 1,000,000 pigs have been slaughtered since 3 August 2018 in China. However, vaccines or drugs for ASF have yet to be developed. As such, a rapid test that can accurately detect ASFV on-site is important to the timely implementation of control measures. In this study, we developed a rapid test that combines recombinase polymerase amplification (RPA) of the ASFV p72 gene with lateral flow detection (LFD). Results showed that the sensitivity of recombinase polymerase amplification with lateral flow dipstick (RPA-LFD) for ASFV was 150 copies per reaction within 10 min at 38°C. The assay was highly specific to ASFV and had no cross-reactions with other porcine viruses, including classical swine fever virus (CSFV). A total of 145 field samples were examined using our method, and the agreement of the positive rate between RPA-LFD (10/145) and real-time PCR (10/145) was 100%. Overall, RPA-LFD provides a novel alternative for the simple, sensitive, and specific identification of ASFV and showed potential for on-site ASFV detection.

摘要

非洲猪瘟病毒(ASFV)是家猪出血热——非洲猪瘟(ASF)的病原体,对养猪业造成了毁灭性后果。自2018年8月3日以来,中国已屠宰了100多万头猪。然而,针对非洲猪瘟的疫苗或药物尚未研发出来。因此,一种能够在现场准确检测ASFV的快速检测方法对于及时实施防控措施至关重要。在本研究中,我们开发了一种将ASFV p72基因的重组酶聚合酶扩增(RPA)与侧向流动检测(LFD)相结合的快速检测方法。结果表明,在38°C下10分钟内,用于检测ASFV的重组酶聚合酶扩增侧向流动试纸条(RPA-LFD)的灵敏度为每个反应150个拷贝。该检测方法对ASFV具有高度特异性,与包括经典猪瘟病毒(CSFV)在内的其他猪病毒无交叉反应。使用我们的方法共检测了145份现场样本,RPA-LFD(10/145)与实时荧光定量PCR(10/145)的阳性率一致性为100%。总体而言,RPA-LFD为简单、灵敏且特异的ASFV鉴定提供了一种新的替代方法,并显示出在现场检测ASFV的潜力。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/232c/6530510/68527f41795b/fmicb-10-01004-g001.jpg

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