A recombinase polymerase amplification combined with lateral flow dipstick for rapid and specific detection of African swine fever virus

被引：27

作者：

Zhai, Yaru ^{[1
]}

Ma, Peng ^{[1
]}

Fu, Xue ^{[1
]}

Zhang, Lan ^{[1
]}

Cui, Pengfei ^{[1
]}

Li, Hao ^{[1
]}

Yan, Wenjun ^{[1
]}

Wang, Hongning ^{[1
]}

Yang, Xin ^{[1
]}

机构：

[1] Sichuan Univ, Anim Dis Prevent & Food Safety Key Lab Sichuan Pr, Key Lab Bioresource & Ecoenvironm, Minist Educ,Coll Life Sci, Chengdu 610065, Sichuan, Peoples R China

来源：

JOURNAL OF VIROLOGICAL METHODS | 2020年 / 285卷

关键词：

African swine fever virus; RPA; Lateral flow dipstick; Rapid detection; PCR ASSAY; DIFFERENTIATION; BLOOD; PIG;

D O I：

10.1016/j.jviromet.2020.113885

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

African swine fever (ASF) is an acute, hemorrhagic, highly contagious disease caused by African swine fever virus (ASFV) infection of domestic pigs and wild boars, showing mortality rates up to 100 %. There are no effective vaccines or antiviral drugs available for ASFV. Therefore, disease control is mainly based on animal slaughtering and the enforcement of strict sanitary measures. In order to establish a rapid, sensitive and simple method for on-site detection of ASFV, a recombinase polymerase amplification (RPA) combined with lateral flow dipstick (LFD) was developed using a pair of specific primers and probe. Using recombinant plasmid pMD19-T-K205R DNA as a template, the RPA-LFD detection could be accomplished in 10 min at a temperature of 36 degrees C-44 degrees C. More specific than PCR and more rapid and simpler than real-time PCR, RPA-LFD has the same detection limit of 1 x 10(2) copies/reaction as real-time PCR, also with no cross-reaction with other viral strains. A convenient and rapid ASFV RPA-LFD detection method was developed, which will provide an efficient method for investigating epidemiology of ASFV infection.

引用

页数：6

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