Development of a Recombinase-aided Amplification Combined With Lateral Flow Dipstick Assay for the Rapid Detection of the African Swine Fever Virus

被引：15

作者：

Li Jiang Shuai ^{[1
]}

Hao Yan Zhe ^{[2
]}

Hou Mei Ling ^{[2
]}

Zhang Xuan ^{[2
]}

Zhang Xiao Guang ^{[2
]}

Cao Yu Xi ^{[3
]}

Li Jin Ming ^{[4
]}

Ma Jing ^{[2
]}

Zhou Zhi Xiang ^{[1
]}

机构：

[1] Beijing Univ Technol, Fac Environm & Life, Beijing 100024, Peoples R China

[2] Chinese Ctr Dis Control & Prevent, Natl Inst Viral Dis Control & Prevent, State Key Lab Infect Dis Prevent & Control, Beijing 102206, Peoples R China

[3] Natl Inst Viral Dis Control & Prevent, Beijing 100052, Peoples R China

[4] China Anim Hlth & Epidemiol Ctr, Qingdao 266032, Shandong, Peoples R China

来源：

BIOMEDICAL AND ENVIRONMENTAL SCIENCES | 2022年 / 35卷 / 02期

基金：

中国国家自然科学基金; 国家重点研发计划;

关键词：

African swine fever virus; Recombinase aided amplification; Lateral flow detection; PCR; DIAGNOSIS; DIFFERENTIATION; CONTACT; PIGS; ASFV;

D O I：

10.3967/bes2022.018

中图分类号：

X [环境科学、安全科学];

学科分类号：

08 ; 0830 ;

摘要：

Objective To establish a sensitive, simple and rapid detection method for African swine fever virus (ASFV) B646L gene. Methods A recombinase-aided amplification-lateral flow dipstick (RAA-LFD) assay was developed in this study. Recombinase-aided amplification (RAA) is used to amplify template DNA, and lateral flow dipstick (LFD) is used to interpret the results after the amplification is completed. The lower limits of detection and specificity of the RAA assay were verified using recombinant plasmid and pathogenic nucleic acid. In addition, 30 clinical samples were tested to evaluate the performance of the RAA assay. Results The RAA-LFD assay was completed within 15 min at 37 degrees C, including 10 min for nucleic acid amplification and 5 minutes for LFD reading results. The detection limit of this assay was found to be 200 copies per reaction. And there was no cross-reactivity with other swine viruses. Conclusion A highly sensitive, specific, and simple RAA-LFD method was developed for the rapid detection of the ASFV.

引用

页码：133 / 140

页数：8

共 23 条

[1] Highly sensitive PCR assay for routine diagnosis of African swine fever virus in clinical samples [J].

Agüero, M ;

Fernández, J ;

Romero, L ;

Mascaraque, CS ;

Arias, M ;

Sánchez-Vizcaíno, JM .

JOURNAL OF CLINICAL MICROBIOLOGY, 2003, 41 (09) :4431-4434

[2] Genotyping field strains of African swine fever virus by partial p72 gene characterisation [J].

Bastos, ADS ;

Penrith, ML ;

Crucière, C ;

Edrich, JL ;

Hutchings, G ;

Roger, F ;

Couacy-Hymann, E ;

Thomson, GR .

ARCHIVES OF VIROLOGY, 2003, 148 (04) :693-706

[3] A multiplex reverse transcription PCR and automated electronic microarray assay for detection and differentiation of seven viruses affecting swine [J].

Erickson, A. ;

Fisher, M. ;

Furukawa-Stoffer, T. ;

Ambagala, A. ;

Hodko, D. ;

Pasick, J. ;

King, D. P. ;

Nfon, C. ;

Polo, R. Ortega ;

Lung, O. .

TRANSBOUNDARY AND EMERGING DISEASES, 2018, 65 (02) :e272-e283

[4] Development of cross-priming amplification for direct detection of the African Swine Fever Virus, in pig and wild boar blood and sera samples [J].

Fraczyk, M. ;

Wozniakowski, G. ;

Kowalczyk, A. ;

Niemczuk, K. ;

Pejsak, Z. .

LETTERS IN APPLIED MICROBIOLOGY, 2016, 62 (05) :386-391

[5] African Swine Fever Virus: A Review [J].

Galindo, Inmaculada ;

Alonso, Covadonga .

VIRUSES-BASEL, 2017, 9 (05)

[6] African swine fever (ASF) diagnosis, an essential tool in the epidemiological investigation [J].

Gallardo, C. ;

Fernandez-Pinero, J. ;

Arias, M. .

VIRUS RESEARCH, 2019, 271

[7] Development and Validation of a Multiplex, Real-Time RT PCR Assay for the Simultaneous Detection of Classical and African Swine Fever Viruses [J].

Haines, Felicity J. ;

Hofmann, Martin A. ;

King, Donald P. ;

Drew, Trevor W. ;

Crooke, Helen R. .

PLOS ONE, 2013, 8 (07)

[8] Rapid detection of infectious bovine Rhinotracheitis virus using recombinase polymerase amplification assays [J].

Hou, Peili ;

Wang, Hongmei ;

Zhao, Guimin ;

He, Chengqiang ;

He, Hongbin .

BMC VETERINARY RESEARCH, 2017, 13

[9] Visual and equipment-free reverse transcription recombinase polymerase amplification method for rapid detection of foot-and-mouth disease virus [J].

Liu, Libing ;

Wang, Jinfeng ;

Zhang, Ruoxi ;

Lin, Mi ;

Shi, Ruihan ;

Han, Qingan ;

Wang, Jianchang ;

Yuan, Wanzhe .

BMC VETERINARY RESEARCH, 2018, 14

[10] Overcoming the challenges of pen-side molecular diagnosis of African swine fever to support outbreak investigations under field conditions [J].

Liu, Lihong ;

Atim, Stella ;

LeBlanc, Neil ;

Rauh, Rolf ;

Esau, Martin ;

Chenais, Erika ;

Mwebe, Robert ;

Nelson, William M. ;

Masembe, Charles ;

Nantima, Noelina ;

Ayebazibwe, Chrissostom ;

Stahl, Karl .

TRANSBOUNDARY AND EMERGING DISEASES, 2019, 66 (02) :908-914

← 1 2 3 →