title: Reverse-transcription loop-mediated isothermal amplification (RT-LAMP) assay for the visual detection of European and North American porcine reproductive and respiratory syndrome viruses.
abstract: A reverse transcription loop-mediated isothermal amplification (RT-LAMP) assay for visual detection of European (EU) and North American (NA) porcine reproductive and respiratory syndrome viruses (PRRSVs) were established and evaluated with reference PRRSV strains and clinical samples. The assay was performed in two reaction tubes containing each set of primers specific for EU or NA-PRRSV at 58°C for 40min, and the results could be visually detected by the naked eye, using hydroxynaphthol blue dye. The detection limit of the assay was 1 or 0.1 TCID50/0.1mL for EU or NA PRRSV, respectively, which was comparable to that of the previously described real-time RT-PCR (qRT-PCR). The detection rate of the assay on 130 field samples was 72.3%, relatively higher than that of qRT-PCR (70.8%), and there was high overall percentage agreement between the two assays. The high specificity, sensitivity, and reliability of the RT-LAMP assay described in this study renders it useful for the rapid and differential diagnosis of EU and NA PRRSVs, even in under-equipped laboratories.
authors.affiliation: College of Veterinary Medicine & Animal Disease Intervention Center, Kyungpook National University, Daegu 41566, Republic of Korea; Animal Disease Diagnostic Division, Animal and Plant Quarantine Agency, Gimcheon 39660, Republic of Korea.
authors.forename: Ji-Young
authors.initials: JY
authors.lastname: Park
authors.affiliation: College of Veterinary Medicine & Animal Disease Intervention Center, Kyungpook National University, Daegu 41566, Republic of Korea.
authors.forename: Soojin
authors.initials: S
authors.lastname: Park
authors.affiliation: College of Veterinary Medicine & Animal Disease Intervention Center, Kyungpook National University, Daegu 41566, Republic of Korea.
authors.forename: Yu-Ri
authors.initials: YR
authors.lastname: Park
authors.affiliation: College of Veterinary Medicine & Animal Disease Intervention Center, Kyungpook National University, Daegu 41566, Republic of Korea; Animal Disease Diagnostic Division, Animal and Plant Quarantine Agency, Gimcheon 39660, Republic of Korea.
authors.forename: Dae-Young
authors.initials: DY
authors.lastname: Kang
authors.affiliation: College of Veterinary Medicine & Animal Disease Intervention Center, Kyungpook National University, Daegu 41566, Republic of Korea; RAD Inc., Daegu 41566, Republic of Korea.
authors.forename: Eun-Mi
authors.initials: EM
authors.lastname: Kim
authors.affiliation: M monitor Inc., Daegu 41944, Republic of Korea.
authors.forename: Hyo-Sung
authors.initials: HS
authors.lastname: Jeon
authors.affiliation: M monitor Inc., Daegu 41944, Republic of Korea.
authors.forename: Ji-Jung
authors.initials: JJ
authors.lastname: Kim
authors.affiliation: Laboratory of Veterinary Immunology, College of Veterinary Medicine, Chonbuk National University, Iksan 54596, Republic of Korea.
authors.forename: Won-Il
authors.initials: WI
authors.lastname: Kim
authors.affiliation: Animal Genomics & Bioinformatics Division, National Institute of Animal Science, Rural Development Administration, Wanju 55365, Republic of Korea.
authors.forename: Kyung-Tai
authors.initials: KT
authors.lastname: Lee
authors.affiliation: Animal Disease Diagnostic Division, Animal and Plant Quarantine Agency, Gimcheon 39660, Republic of Korea.
authors.forename: Seong-Hee
authors.initials: SH
authors.lastname: Kim
authors.affiliation: Animal Disease Diagnostic Division, Animal and Plant Quarantine Agency, Gimcheon 39660, Republic of Korea.
authors.forename: Kyoung-Ki
authors.initials: KK
authors.lastname: Lee
authors.affiliation: College of Veterinary Medicine & Animal Disease Intervention Center, Kyungpook National University, Daegu 41566, Republic of Korea. Electronic address: parkck@knu.ac.kr.
authors.forename: Choi-Kyu
authors.initials: CK
authors.lastname: Park
mesh_terms: D000818:Animals; D004396:Coloring Agents; D017931:DNA Primers; D005060:Europe; D057230:Limit of Detection; D021141:Nucleic Acid Amplification Techniques / Q000379:methods*; D019318:Porcine Reproductive and Respiratory Syndrome / Q000175:diagnosis* / Q000821:virology; D019316:Porcine respiratory and reproductive syndrome virus / Q000302:isolation & purification* / Q000648:ultrastructure; D060888:Real-Time Polymerase Chain Reaction / Q000379:methods; D015203:Reproducibility of Results; D048348:Reverse Transcription; D012680:Sensitivity and Specificity; D013552:Swine; D013696:Temperature; D014481:United States
chemical_list: D004396:Coloring Agents; D017931:DNA Primers
keywords: PRRSV; RT-LAMP; Real-time RT-PCR