Rapid Visual Detection of African Swine Fever Virus with a CRISPR/Cas12a Lateral Flow Strip Based on Structural Protein Gene D117L

被引：9

作者：

Zhang, Desheng ^{[1
,2
,3
,4
]}

Jiang, Sen ^{[1
,2
,3
,4
]}

Xia, Nengwen ^{[1
,2
,3
,4
]}

Zhang, Youwen ^{[1
,2
,3
,4
]}

Zhang, Jiajia ^{[1
,2
,3
,4
]}

Liu, Anjing ^{[1
,2
,3
,4
]}

Zhang, Chenyang ^{[1
,2
,3
,4
]}

Chen, Nanhua ^{[1
,2
,3
,4
]}

Meurens, Francois ^{[5
,6
]}

Zheng, Wanglong ^{[1
,2
,3
,4
]}

Zhu, Jianzhong ^{[1
,2
,3
,4
]}

机构：

[1] Yangzhou Univ, Coll Vet Med, Yangzhou 225009, Peoples R China

[2] Joint Int Res Lab Agr & Agriprod Safety, Yangzhou 225009, Peoples R China

[3] Yangzhou Univ, Comparat Med Res Inst, Yangzhou 225009, Peoples R China

[4] Yangzhou Univ, Jiangsu Coinnovat Ctr Prevent & Control Important, Yangzhou 225009, Peoples R China

[5] Univ Montreal, Fac Vet Med, Swine & Poultry Infect Dis Res Ctr, St Hyacinthe, PQ J2S 2M2, Canada

[6] Univ Saskatchewan, Western Coll Vet Med, Dept Vet Microbiol & Immunol, Saskatoon, SK S7N 5E2, Canada

来源：

ANIMALS | 2023年 / 13卷 / 23期

关键词：

African swine fever virus (ASFV); D117L; Cas12a; lateral flow strip (LFS); sensitivity and specificity; detection; AMPLIFICATION;

D O I：

10.3390/ani13233712

中图分类号：

S8 [畜牧、动物医学、狩猎、蚕、蜂];

学科分类号：

0905 ;

摘要：

Simple Summary African swine fever, caused by African swine fever virus, seriously affects domestic pigs and wild boars and has brought huge economic losses to endemic countries and regions. However, there is still a lack of effective vaccines and therapeutics at present. The prevention and control of African swine fever largely depends on early and accurate detection, and thus the desired detection method of clinical African swine fever virus is indeed needed. Despite a number of detection methods available, we here develop a lateral flow strip detection method for African swine fever virus, based on the viral gene of structural protein named p17 and the new technology called the clustered regularly interspaced short palindromic repeats/Cas 12a nuclease system. This visual detection method is simple, rapid, sensitive, and specific and has the potential for clinical detection of African swine fever virus.Abstract African swine fever virus (ASFV) is a large double-stranded DNA virus that is highly infectious and seriously affects domestic pigs and wild boars. African swine fever (ASF) has caused huge economic losses to endemic countries and regions. At present, there is still a lack of effective vaccines and therapeutics. Therefore, rapid and accurate detection is essential for the prevention and control of ASF. The portable DNA endonuclease (Cas12a)-mediated lateral flow strip detection method (Cas12a-LFS) combined with recombinant polymerase amplification (RPA) has been gradually recognized as effective for virus detection including ASFV. In this study, based on the ASFV structural protein p17 gene (D117L), an RPA-Cas12a-LFS detection method was established. The detection method exhibits a sensitivity of up to two gene copies and has no cross-reaction with nine other swine viruses. Thus, the method is highly sensitive and specific. In 68 clinical samples, the coincidence rate of the p17 strip was 100%, compared to the traditional quantitative PCR (qPCR). In conclusion, we have developed a simple, rapid, sensitive, and specific ASFV visual detection method and demonstrated the potential of on-site detection of ASFV.

引用

页数：13

共 37 条

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