title: Specific detection of H5N1 avian influenza A virus in field specimens by a one-step RT-PCR assay.
abstract: BACKGROUND
  Continuous outbreaks of the highly pathogenic H5N1 avian influenza A in Asia has resulted in an urgent effort to improve current diagnostics to aid containment of the virus and lower the threat of a influenza pandemic. We report here the development of a PCR-based assay that is highly specific for the H5N1 avian influenza A virus.
  
  
  METHODS
  A one-step reverse-transcription PCR assay was developed to detect the H5N1 avian influenza A virus. The specificity of the assay was shown by testing sub-types of influenza A virus and other viral and bacterial pathogens; and on field samples.
  
  
  RESULTS
  Validation on 145 field specimens from Vietnam and Malaysia showed that the assay was specific without cross reactivity to a number of other infuenza strains as well as human respiratory related pathogens. Detection was 100% from allantoic fluid in H5N1 positive samples, suggesting it to be a reliable sampling source for accurate detection.
  
  
  CONCLUSIONS
  The assay developed from this study indicates that the primers are specific for the H5N1 influenza virus. As shown by the field tested results, this assay would be highly useful as a diagnostic tool to help identify and control influenza epidemics.
authors.affiliation: Genome Institute of Singapore, 60 Biopolis Street, Genome, #02-01, 138672, Singapore. ngfp@gis.a-star.edu.sg
authors.forename: Lisa Fp
authors.initials: LF
authors.lastname: Ng
authors.forename: Ian
authors.initials: I
authors.lastname: Barr
authors.forename: Tung
authors.initials: T
authors.lastname: Nguyen
authors.forename: Suriani Mohd
authors.initials: SM
authors.lastname: Noor
authors.forename: Rosemary Sok-Pin
authors.initials: RS
authors.lastname: Tan
authors.forename: Lora V
authors.initials: LV
authors.lastname: Agathe
authors.forename: Sanjay
authors.initials: S
authors.lastname: Gupta
authors.forename: Hassuzana
authors.initials: H
authors.lastname: Khalil
authors.forename: Thanh Long
authors.initials: TL
authors.lastname: To
authors.forename: Sharifah Syed
authors.initials: SS
authors.lastname: Hassan
authors.forename: Ee-Chee
authors.initials: EC
authors.lastname: Ren
mesh_terms: D003429:Cross Reactions; D006801:Humans; D053124:Influenza A Virus, H5N1 Subtype / Q000235:genetics* / Q000302:isolation & purification*; D015203:Reproducibility of Results; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods*; D012680:Sensitivity and Specificity