title: Specific, simple and rapid detection of porcine circovirus type 2 using the loop-mediated isothermal amplification method.
abstract: BACKGROUND
  Porcine circovirus type 2 (PCV2) is the causative agent of postweaning multisystemic wasting syndrome (PMWS), and porcine dermatitis and nephropathy syndrome (PDNS). It has caused heavy losses in global agriculture in recent decades. Rapid detection of PCV2 is very important for the effective prophylaxis and treatment of PMWS.
  
  
  RESULTS
  A loop-mediated isothermal amplification (LAMP) assay was used to detect PCV2 in this study. Three pairs of primers were specially designed for recognizing eight distinct sequences of the ORF2 gene. This gene lies in the PCV2 virus genome sequence, and encodes the Rep protein that is involved in virus replication. Time and temperature conditions for amplification of PCV2 genes were optimized to be 55 min at 59°C. The analysis of clinical samples indicated that the LAMP method was highly sensitive. The detection limit for PCV2 by the LAMP assay was 10 copies, whereas the limit by conventional PCR was 1000 copies. The assay did not cross-react with PCV1, porcine reproductive and respiratory syndrome virus, porcine epidemic diarrhea virus, transmissible gastroenteritis of pigs virus or rotavirus. When 110 samples were tested using the established LAMP system, 95 were detected as positive.
  
  
  CONCLUSIONS
  The newly developed LAMP detection method for PCV2 was more specific, sensitive, rapid and simple than before. It complements and extends previous methods for PCV2 detection and provides an alternative approach for detection of PCV2.
authors.affiliation: Biotechnology Research Institute, Shanghai Academy of Agricultural Sciences, 2901 Beidi Road, Shanghai, 201106, People's Republic of China.
authors.forename: Kai
authors.initials: K
authors.lastname: Zhao
authors.forename: Wei
authors.initials: W
authors.lastname: Shi
authors.forename: Fangting
authors.initials: F
authors.lastname: Han
authors.forename: Yan
authors.initials: Y
authors.lastname: Xu
authors.forename: Lianlong
authors.initials: L
authors.lastname: Zhu
authors.forename: Yong
authors.initials: Y
authors.lastname: Zou
authors.forename: Xiao
authors.initials: X
authors.lastname: Wu
authors.forename: Hong
authors.initials: H
authors.lastname: Zhu
authors.forename: Furong
authors.initials: F
authors.lastname: Tan
authors.forename: Shiru
authors.initials: S
authors.lastname: Tao
authors.forename: Xueming
authors.initials: X
authors.lastname: Tang
mesh_terms: D000818:Animals; D017925:Circovirus / Q000235:genetics / Q000302:isolation & purification*; D021141:Nucleic Acid Amplification Techniques / Q000379:methods*; D053570:Porcine Postweaning Multisystemic Wasting Syndrome / Q000175:diagnosis / Q000821:virology*; D012680:Sensitivity and Specificity; D013552:Swine; D014764:Viral Proteins / Q000235:genetics
chemical_list: D014764:Viral Proteins