title: Development of a loop-mediated isothermal amplification assay for rapid detection of bovine parvovirus.
abstract: A loop-mediated isothermal amplification (LAMP) assay was developed for detection of bovine parvovirus (BPV) DNA. Four primers were designed to recognize six distinct regions on the target DNA based on a highly conserved sequence in the VP2 region of the BPV genome. The optimized LAMP reaction conditions were 8 mM Mg²⁺, 1.2 mM betaine, and an incubation at 63°C for 45 min. After amplification the products were detected either by observing a ladder pattern following gel electrophoresis, observation of turbidity, or a color change with the addition of SYBR Green I to the reaction tube. The detection limit of the LAMP assay was 9 copies of BPV-DNA and was 100 times more sensitive than conventional PCR. A ladder pattern of bands after gel electrophoresis was observed for only BPV isolates and showed that the BPV LAMP assay was highly specific without any cross-reactivity with other related viruses. The LAMP assay was evaluated further using 59 field samples and the results were comparable to conventional PCR. The LAMP assay is a simple, rapid and economic detection method; it can provide a useful technique suitable for detection of BPV infection in both field conditions and laboratory settings.
authors.affiliation: College of Veterinary Medicine, Northeast Agricultural University, Harbin 150030, China.
authors.forename: Ji-guan
authors.initials: JG
authors.lastname: Luo
authors.forename: Jun-wei
authors.initials: JW
authors.lastname: Ge
authors.forename: Li-jie
authors.initials: LJ
authors.lastname: Tang
authors.forename: Xin-yuan
authors.initials: XY
authors.lastname: Qiao
authors.forename: Yan-ping
authors.initials: YP
authors.lastname: Jiang
authors.forename: Wen
authors.initials: W
authors.lastname: Cui
authors.forename: Min
authors.initials: M
authors.lastname: Liu
authors.forename: Yi-jing
authors.initials: YJ
authors.lastname: Li
mesh_terms: D000818:Animals; D052660:Bocavirus / Q000235:genetics / Q000302:isolation & purification*; D002417:Cattle; D002418:Cattle Diseases / Q000175:diagnosis* / Q000821:virology; D017931:DNA Primers / Q000235:genetics; D004279:DNA, Viral / Q000235:genetics; D025202:Molecular Diagnostic Techniques / Q000379:methods*; D021141:Nucleic Acid Amplification Techniques / Q000379:methods*; D010322:Parvoviridae Infections / Q000662:veterinary* / Q000821:virology; D012680:Sensitivity and Specificity; D014730:Veterinary Medicine / Q000379:methods*
chemical_list: D017931:DNA Primers; D004279:DNA, Viral