title: Loop-mediated isothermal amplification for detection of porcine circovirus type 2.
abstract: BACKGROUND
  Porcine circovirus type 2 (PCV2) is the primary causative agent of the emerging swine disease known as postweaning multisystemic wasting syndrome (PMWS). Nowadays, polymerase chain reaction (PCR) is still the most widespread technique in pathogen detection. Loop-mediated isothermal amplification (LAMP), a novel nucleic acid amplification method developed in 2000, will possibly replace PCR in the field of detection. To establish a LAMP method for rapid detection of PCV2, two pairs of primers were designed specially from the open reading frame 2 (ORF2) sequences of PCV2. A LAMP method for rapid detection of PCV2 was established. To compare with PCR, sensitivity and specificity of LAMP were evaluated using the optimized reaction system. The LAMP products could be determined by agarose gel electrophoresis or adding SYBR Green I dye.
  
  
  RESULTS
  The amplification of LAMP could be obtained at 63°C for 60 min. The detection limit was nearly 1 copy of DNA plasmid, more sensitive than PCR. There was no cross-reaction with porcine circovirus type 1 (PCV1), porcine pseudorabies virus (PRV) and porcine parvovirus (PPV) under the same conditions.
  
  
  CONCLUSIONS
  LAMP is an useful rapid detection method with high sensitivity and specificity for PCV2.
authors: Qingdao Agricultural University, Qingdao, 266109, China.
authors: Shun
authors: S
authors: Zhou
authors: Si
authors: S
authors: Han
authors: Jianli
authors: J
authors: Shi
authors: Jiaqiang
authors: J
authors: Wu
authors: Xiaoyuan
authors: X
authors: Yuan
authors: Xiaoyan
authors: X
authors: Cong
authors: Shaojian
authors: S
authors: Xu
authors: Xiaoyan
authors: X
authors: Wu
authors: Jun
authors: J
authors: Li
authors: Jinbao
authors: J
authors: Wang
mesh_terms: D000818:Animals; D052160:Benzothiazoles; D018173:Circoviridae Infections / Q000175:diagnosis / Q000662:veterinary* / Q000821:virology; D017925:Circovirus / Q000302:isolation & purification*; D017931:DNA Primers / Q000235:genetics; D003959:Diamines; D004587:Electrophoresis, Agar Gel; D021141:Nucleic Acid Amplification Techniques / Q000379:methods*; D016366:Open Reading Frames; D009930:Organic Chemicals / Q000378:metabolism; D011804:Quinolines; D012680:Sensitivity and Specificity; D013194:Staining and Labeling; D013552:Swine; D013553:Swine Diseases / Q000175:diagnosis* / Q000821:virology*; D014773:Virology / Q000379:methods*
chemical_list: D052160:Benzothiazoles; D017931:DNA Primers; D003959:Diamines; D009930:Organic Chemicals; D011804:Quinolines; C098022:SYBR Green I