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http://dx.doi.org/10.7853/kjvs.2022.45.1.19

Simple and rapid colorimetric detection of African swine fever virus by loop-mediated isothermal amplification assay using a hydroxynaphthol blue metal indicator  

Park, Ji-Hoon (College of Veterinary Medicine & Animal Disease Intervention Center, Kyungpook National University)
Kim, Hye-Ryung (College of Veterinary Medicine & Animal Disease Intervention Center, Kyungpook National University)
Chae, Ha-Kyung (College of Veterinary Medicine & Animal Disease Intervention Center, Kyungpook National University)
Park, Jonghyun (College of Veterinary Medicine & Animal Disease Intervention Center, Kyungpook National University)
Jeon, Bo-Young (Department of Biomedical Laboratory Science, College of Health Science, Yonsei University)
Lyoo, Young S. (College of Veterinary Medicine, Konkuk University)
Park, Choi-Kyu (College of Veterinary Medicine & Animal Disease Intervention Center, Kyungpook National University)
Publication Information
Korean Journal of Veterinary Service / v.45, no.1, 2022 , pp. 19-30 More about this Journal
Abstract
In this study, a simple loop-mediated isothermal amplification (LAMP) combined with visual detection method (vLAMP) assay was developed for the rapid and specific detection of African swine fever virus (ASFV), overcoming the shortcomings of previously described LAMP assays that require additional detection steps or pose a cross-contamination risk. The assay results can be directly detected by the naked eye using hydroxynaphthol blue after incubation for 40 min at 62℃. The assay specifically amplified ASFV DNA and no other viral nucleic acids. The limit of detection of the assay was <50 DNA copies/reaction, which was ten times more sensitive than conventional polymerase chain reaction (cPCR) and comparable to real-time PCR (qPCR). For clinical evaluation, the ASFV detection rate of vLAMP was higher than cPCR and comparable to OIE-recommended qPCR, showing 100% concordance, with a κ value (95% confidence interval) of 1 (1.00~1.00). Considering the advantages of high sensitivity and specificity, no possibility for cross-contamination, and being able to be used as low-cost equipment, the developed vLAMP assay will be a valuable tool for detecting ASFV from clinical samples, even in resource-limited laboratories.
Keywords
African swine fever virus; Loop-mediated isothermal amplification; Visual detection;
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1 Yoo DS, Kim Y, Lee ES, Lim JS, Hong SK, Lee IS, Jung CS, Yoon HC, Wee SH, Pfeiffer DU, Fournie G. 2021. Transmission dynamics of African swine fever virus, South Korea, 2019. Emerg Infect Dis. 27(7): 1909-1918.   DOI
2 Aguero M, Fernadez J, Romero L, Sanchez C, Arias M, Sanchez-Vizcaino JM. 2003. Highly sensitive PCR assay for the routine diagnosis of African swine fever virus in clinical samples. J Clin Microbiol. 41(9): 4431-4434.   DOI
3 Alonso C, Borca M, Dixon L, Revilla Y, Rodriguez F, Escribano JM, ICTV Report Consortium. 2018. ICTV virus taxonomy profile: Asfarviridae. J Gen Virol. 99: 613-614.   DOI
4 Atuhaire DK, Afayoa M, Ochwo S, Katiti D, Mwiine FN, Nanteza A, Mugasa CM, Matovu E, Okuni JB, Olaho-Mukani W, Ojok L. 2014. Comparative detection of African swine fever virus by loop-mediated isothermal amplification assay and polymerase chain reaction in domestic pigs in Uganda. African J Microbiol Res. 8(23): 2322-2328.   DOI
5 Dhama K, Karthik K, Chakraborty S, Tiwari R, Kapoor S, Kumar A, Thomas P. 2014. Loop-mediated isothermal amplification of DNA (LAMP): a new diagnostic tool lights the world of diagnosis of animal and human pathogens: a review. Pak J Biol Sci. 17: 151-166.   DOI
6 Gallardo C, Fernandez-Pinero J, Arias M. 2019. African swine fever (ASF) diagnosis, an essential tool in the epidemiological investigation. Virus Res. 271: 197676.   DOI
7 James HE, Ebert K, McGonigle R, Reid SM, Boonham N, Tomlinson JA, Hutchings GH, Denyer M, Oura CA, Dukes JP, King DP. 2010. Detection of African swine fever virus by loop-mediated isothermal amplification. J Virol Methods 164(1-2): 68-74.   DOI
8 Jo YS, Gortazar C. 2021. African swine fever in wild boar: Assessing interventions in South Korea. Transbound Emerg Dis. 68(5): 2878-2889.   DOI
9 Kim HJ, Cho KH, Lee SK, Kim DY, Nah JJ, Kim HJ, Kim HJ, Hwang JY, Sohn HJ, Choi JG, Kang HE, Kim YJ. 2020a. Outbreak of African swine fever in South Korea, 2019. Transbound Emerg Dis. 67(2): 473-475.   DOI
10 Quembo CJ, Jori F, Vosloo W, Heath L. 2018. Genetic characterization of African swine fever virus isolates from soft ticks at the wildlife/domestic interface in Mozambique and identification of a novel genotype. Transbound Emerg Dis. 65(2): 420-431.   DOI
11 Bastos AD, Penrith ML, Cruciere C, Edrich JL, Hutchings G, Roger F, Couacy-Hymann E, Thomson GR. 2003. Genotyping field strains of African swine fever virus by partial p72 gene characterisation. Arch Virol. 148(4): 693-706.   DOI
12 Kim JK, Kim HR, Kim DY, Kim JM, Kwon NY, Park JH, Park JY, Kim SH, Lee KK, Lee C, Joo HD, Lyoo YS, Park CK. 2021. A simple colorimetric detection of porcine epidemic diarrhea virus by reverse transcription loop-mediated isothermal amplification assay using hydroxynaphthol blue metal indicator. J Virol Methods 298: 114289.   DOI
13 Kim DY, Kim HR, Park JH, Kwon NY, Kim JM, Kim JK, Park JH, Lee KK, Kim SH, Kim WI, Lyoo YS, Park CK. 2022. Detection of a novel porcine circovirus 4 in Korean pig herds using a loop-mediated isothermal amplification assay. J Virol Methods 299: 114350.   DOI
14 Wang D, Yu J, Wang Y, Zhang M, Li P, Liu M, Liu Y. 2020. Development of a real-time loop-mediated isothermal amplification (LAMP) assay and visual LAMP assay for detection of African swine fever virus (ASFV). J Virol Methods 276: 113775.   DOI
15 Zsak L, Borca MV, Risati GR, Zsak A, French RA, Lu Z, Kutish GF, Neilan JG, Callahan JD, Nelson WM, Rock DL. 2005. Preclinical diagnosis of African swine fever in contact-exposed swine by a real-Time PCR Assay. J Clin Microbiol. 43(1): 112-119.   DOI
16 Kim HJ, Cho KH, Ryu JH, Jang MK, Chae HG, Choi JD, Nah JJ, Kim YJ, Kang HE. 2020b. Isolation and genetic characterization of African swine fever virus from domestic pig farms in South Korea, 2019. Viruses 12(11): 1237.   DOI
17 Kim HR, Park YR, Lim DR, Park MJ, Park JY, Kim SH, Lee KK, Lyoo YS, Park CK. 2017. Multiplex realtime polymerase chain reaction for the differential detection of porcine circovirus 2 and 3. J Virol Methods 250: 11-16.   DOI
18 Kwiecien R, Kopp-Schneider A, Blettner M. 2011. Concordance analysis: Part 16 of a series on evaluation of scientific publications. Dtsch Arztebl Int. 108(30): 515-521.
19 Dokphut A, Boonpornprasert P, Songkasupa T, Tangdee S. 2021. Development of a loop-mediated isothermal amplification assay for rapid detection of African swine fever. Vet Integr Sci. 19(1): 87-100.
20 Chae HG, Lim DR, Kim HR, Park MJ, Park CK. 2020. An advanced loop-mediated isothermal amplification assay for the rapid detection of beak and feather disease virus in psittacine birds. J Virol Methods 277: 113819.   DOI
21 Fernandez-Pinero J, Gallardo C, Elizalde M, Robles A, Gomez C, Bishop R, Heath L, Couacy-Hymann E, Fasina FO, Pelayo V, Soler A, Arias M. 2013. Molecular diagnosis of African swine fever by a new real-time PCR using universal probe library. Transbound Emerg Dis. 60(1): 48-58.   DOI
22 Gaudreault NN, Madden DW, Wilson WC, Trujillo JD, Richt JA. 2020. African swine fever virus: an emerging DNA Arbovirus. Front Vet Sci. 7:215.   DOI
23 Goto M, Honda E, Ogura A, Nomoto A, Hanaki KI, 2009. Colorimetric detection of loop-mediated isothermal amplification reaction by using hydroxy naphthol blue. Biotechniques 46: 167-172.   DOI
24 Park YR, Kim HR, Kim SH, Lee KK, Lyoo YS, Yeo SG, Park CK. 2018. Loop-mediated isothermal amplification assay for the rapid and visual detection of novel porcine circovirus 3. J Virol Methods 53: 26-30.
25 Lim DR, Kim HR, Park MJ, Chae HG, Ku BK, Nah JJ, Ryoo SY, Wee SH, Park YR, Jeon HS, Kim JJ, Jeon BY, Lee HW, Yeo SG, Park CK. 2018. An improved reverse transcription loop-mediated isothermal amplification for sensitive and specific detection of serotype O foot-and-mouth disease virus. J Virol Methods 260: 6-13.   DOI
26 Mori Y, Notomi T. 2009. Loop-mediated isothermal amplification (LAMP): a rapid, accurate, and costeffective diagnostic method for infectious diseases. J Infect Chemother. 15: 62-69.   DOI
27 Park CK, Lee KK, Kim HS. 2004. Genetic characterization of porcine circovirus 2 Korean isolates. Korean J Vet Res. 44: 571-579.
28 Wu X, Xiao L, Wang Y, Yang Z, Yao X, Peng B. 2016. Development of a rapid and sensitive method for detection of African swine fever virus using loopmediated isothermal amplification. Brazilian Arch Biol Technol. 59: e16160500.
29 Park MJ, Kim HR, Chae HG, Lim DR, Kwon OD, Cho KH, Park CK. 2019. Development of a colorimetric loop-mediated isothermal amplification assay for rapid and specific detection of Aves polyomavirus 1 from psittacine birds. J Virol Methods 273: 113687.   DOI
30 Tignon M, Gallardo C, Iscaro C, Hutet E, Van der Stede Y, Kolbasov D, De Mia GM, Le Potier MF, Bishop RP, Arias M, Koenen F. 2011. Development and inter-laboratory validation study of an improved new real-time PCR assay with internal control for detection and laboratory diagnosis of African swine fever virus. J Virol Methods. 178(1-2): 161-170.   DOI
31 King DP, Reid SM, Hutchings GH, Grierson SS, Wilkinson PJ, Dixon LK, Bastos AD, Drew TW. 2003. Development of a TaqMan PCR assay with internal amplification control for the detection of African swine fever virus. J Virol Methods. 107(1): 53-61.   DOI
32 Zhou X, Li N, Luo Y, Liu Y, Miao F, Chen T, Zhang S, Cao P, Li X, Tian K, Qiu HJ, Hu R. 2018. Emergence of African swine fever in China, 2018. Transbound Emerg Dis. 65(6): 1482-1484.   DOI
33 Notomi T, Okayama H, Masubuchi H, Yonekawa T, Watanabe K, Amino N, Hase T. 2000. Loop-mediated isothermal amplification of DNA. Nucleic Acids Res. 28: e63.   DOI
34 Wozniakowski G, Fraczyk M, Mazur N. 2018. Comparison of loop-mediated isothermal amplification (LAMP) and cross-priming amplification (CPA) for detection of African swine fever virus. Polish J Vet Sci. 21(4): 827-830.