title: Development of a loop-mediated isothermal amplification method to rapidly detect porcine circovirus genotypes 2a and 2b.
abstract: BACKGROUND
  Porcine circovirus type 2 (PCV2), is nowadays associated with a number of diseases known as porcine circovirus-associated diseases (PCVAD), especially postweaning multisystemic wasting syndrome (PMWS). The epidemiological investigation of PCV2 infection was usually conducted by PCR, nested PCR, PCR-RFLP, TaqMan-based assay and nucleotide sequencing. However, there is still no rapid, sensitive and practical method for detecting PCV2 genotypes. As a novel nucleic acid amplification method, the loop-mediated isothermal amplification method (LAMP) has been used to detect a variety of pathogenic microorganisms.
  
  
  RESULTS
  Herein, a LAMP method is developed to detect the genotypes of PCV2. The diagnostic sensitivity of LAMP is 1 copy/reaction for differentiating genotypes PCV2a and PCV2b. The reaction process was completed at 65°C for 1 hour in a water bath. Cross-reactivity assay shows that this method is specific for PCV2a and PCV2b and no reactive for PCV2c and other swine-origin viruses (i.e. CSFV, PRRSV, BVDV, TGEV and PEDV, etc). Identity between LAMP and nested PCR was 92.3% on 52 field clinical samples.
  
  
  CONCLUSIONS
  LAMP method provides a rapid, sensitive, reliable way to detect PCV2a and PCV2b, and a better means for the large scale investigation of PCV2a and PCV2b infection.
authors.affiliation: Key Laboratory of Animal Virology of Ministry of Agriculture, College of Animal Sciences, Zhejiang University, 866 Yuhangtang Road, Hangzhou 310058, PR China.
authors.forename: Xiaohuo
authors.initials: X
authors.lastname: Qiu
authors.forename: Tian
authors.initials: T
authors.lastname: Li
authors.forename: Guorui
authors.initials: G
authors.lastname: Zhang
authors.forename: Jingjing
authors.initials: J
authors.lastname: Cao
authors.forename: Yulan
authors.initials: Y
authors.lastname: Jin
authors.forename: Gang
authors.initials: G
authors.lastname: Xing
authors.forename: Min
authors.initials: M
authors.lastname: Liao
authors.forename: Jiyong
authors.initials: J
authors.lastname: Zhou
mesh_terms: D000818:Animals; D018173:Circoviridae Infections / Q000175:diagnosis / Q000662:veterinary* / Q000821:virology; D017925:Circovirus / Q000145:classification / Q000235:genetics / Q000302:isolation & purification*; D017931:DNA Primers; D004279:DNA, Viral / Q000235:genetics; D005838:Genotype; D021141:Nucleic Acid Amplification Techniques / Q000379:methods; D016133:Polymerase Chain Reaction; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D013552:Swine; D013553:Swine Diseases / Q000175:diagnosis* / Q000821:virology; D013997:Time Factors
chemical_list: D017931:DNA Primers; D004279:DNA, Viral