title: Rapid and specific detection of H3 swine influenza virus using reverse transcription loop-mediated isothermal amplification method.
abstract: OBJECTIVE
  The main objective of our study is to develop a reverse transcriptase loop-mediated isothermal amplification (RT-LAMP)-based system for rapid and specific detection of H3 swine influenza virus (SIV).
  
  
  RESULTS
  The system, H3 RT-LAMP, contained a set of six novel primers that targeted eight distinct regions of the viral haemagglutinin (HA) gene that are highly conserved among H3 influenza A viruses but not between H3 and other subtypes. H3 RT-LAMP accurately and specifically detected H3 SIV of different isolates from culture and from swine lung samples. The system is at least 10-fold more sensitive than the conventional RT-PCR assay and even comparable to the real-time RT-PCR method, with the detection limit of about one plaque-forming unit per reaction. Of 27 swine lung samples tested, 11 samples were positive in reactions with the RT-LAMP and real-time RT-PCR methods, while only 7 were positive with the conventional RT-PCR assay. Importantly, the assay can be completed within 45 min and is faster than the conventional RT-PCR and real-time RT-PCR approaches.
  
  
  CONCLUSIONS
  Our results provide the first direct evidence that RT-LAMP is highly specific and sensitive for detecting H3 SIV.
  
  
  CONCLUSIONS
  These results suggest that LAMP offers a promising alternative tool for rapid, inexpensive and specific diagnosis of influenza virus infection of swine and other animals in frontline settings.
authors.affiliation: Institute of Virology, State Key Laboratory of Pharmaceutical Biotechnology, School of Life Sciences, Nanjing University, Nanjing, Jiangsu, China.
authors.forename: H
authors.initials: H
authors.lastname: Gu
authors.forename: X
authors.initials: X
authors.lastname: Qi
authors.forename: X
authors.initials: X
authors.lastname: Li
authors.forename: H
authors.initials: H
authors.lastname: Jiang
authors.forename: Y
authors.initials: Y
authors.lastname: Wang
authors.forename: F
authors.initials: F
authors.lastname: Liu
authors.forename: S
authors.initials: S
authors.lastname: Lu
authors.forename: Y
authors.initials: Y
authors.lastname: Yang
authors.forename: F
authors.initials: F
authors.lastname: Liu
mesh_terms: D000818:Animals; D017931:DNA Primers; D009980:Influenza A virus / Q000235:genetics* / Q000302:isolation & purification*; D008168:Lung / Q000821:virology; D021141:Nucleic Acid Amplification Techniques; D009976:Orthomyxoviridae Infections / Q000175:diagnosis / Q000662:veterinary*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012680:Sensitivity and Specificity; D013552:Swine; D013553:Swine Diseases / Q000175:diagnosis
chemical_list: D017931:DNA Primers