title: Rapid and sensitive detection of porcine epidemic diarrhea virus by reverse transcription loop-mediated isothermal amplification combined with a vertical flow visualization strip.
abstract: Porcine epidemic diarrhea virus (PEDV) is an important pathogen that causes vomiting, diarrhea, and dehydration, leading to serious damage to the swine industry worldwide. The establishment of effective diagnostic methods is imperative. However, traditional methods are often unsuitable. In this study, reverse transcription loop-mediated isothermal amplification (RT-LAMP) was combined with a vertical flow (VF) nucleic acid detection strip to detect PEDV. Parameters that affect the RT-LAMP reaction were optimized. The RT-LAMP-VF assay that we established was performed at 62 °C for 40 min, and then directly evaluated on the VF visualization strip cassette. The method demonstrated high specificity for PEDV. The detection limit was 10 pg of ribonucleic acid, consistent with RT-PCR, RT-LAMP detected products on agarose gels and by direct calcein fluorescence. Application of this method to clinical samples yielded a positivity rate that was comparable to that obtained for RT-PCR. This technique saves time and is efficient, and is thus expected to be useful for the diagnosis of PEDV infection in the field.
authors.affiliation: College of Veterinary Medicine, South China Agricultural University, Guangzhou, PR China.
authors.forename: Hongchao
authors.initials: H
authors.lastname: Gou
authors.affiliation: College of Veterinary Medicine, South China Agricultural University, Guangzhou, PR China.
authors.forename: Jieru
authors.initials: J
authors.lastname: Deng
authors.affiliation: College of Veterinary Medicine, South China Agricultural University, Guangzhou, PR China.
authors.forename: Jiaying
authors.initials: J
authors.lastname: Wang
authors.affiliation: College of Veterinary Medicine, South China Agricultural University, Guangzhou, PR China.
authors.forename: Jingjing
authors.initials: J
authors.lastname: Pei
authors.affiliation: College of Veterinary Medicine, South China Agricultural University, Guangzhou, PR China.
authors.forename: Wenjun
authors.initials: W
authors.lastname: Liu
authors.affiliation: College of Veterinary Medicine, South China Agricultural University, Guangzhou, PR China.
authors.forename: Mingqiu
authors.initials: M
authors.lastname: Zhao
authors.affiliation: College of Veterinary Medicine, South China Agricultural University, Guangzhou, PR China. Electronic address: jdchen@scau.edu.cn.
authors.forename: Jinding
authors.initials: J
authors.lastname: Chen
mesh_terms: D000818:Animals; D018352:Coronavirus Infections / Q000175:diagnosis* / Q000821:virology; D025202:Molecular Diagnostic Techniques / Q000379:methods*; D021141:Nucleic Acid Amplification Techniques / Q000379:methods*; D053485:Porcine epidemic diarrhea virus / Q000145:classification / Q000302:isolation & purification*; D012367:RNA, Viral / Q000032:analysis; D048348:Reverse Transcription; D012680:Sensitivity and Specificity; D013552:Swine; D013553:Swine Diseases / Q000175:diagnosis / Q000821:virology*
chemical_list: D012367:RNA, Viral
keywords: Porcine epidemic diarrhea virus; Rapid detection; Reverse transcription loop-mediated isothermal amplification; Vertical flow visualization strip