title: Methods for molecular surveillance of influenza used in Macedonia.
abstract: OBJECTIVE
  To present and compare different Nucleic Acid Testing assays used for laboratory diagnosis of influenza virus infection in our country.
  
  
  METHODS
  Respiratory samples used were nose and throat swabs. The RNA extraction was performed with a QIAamp viral RNA kit. During the season 2009–2010 the first 25 samples were tested with: conventional gel-based RT-PCR and CDC rtRT-PCR using published specific matrix and HA gene primers and probes for influenza virus typing and subtyping.
  
  
  RESULTS
  Of 25 samples tested with conventional RT-PCR 7(28%) were positive for influenza A, but negative for A/H1seasonal and A/H3. Retested with rtRT-PCR 9(36%) were positive for influenza A, 8(32%) were positive for А/H1pdm and 1(4%) was А/H3. Two samples positive with rtRT-PCR for influenza A were negative with RT-PCR. The sensitivity of the RT-PCR in comparison with rtRT-PCR is 100% and the specificity is 88.89%. Positive predictive value for RT-PCR is 77.78%, and negative predictive value is 100%. RT-PCR is a four-step and rtRT-PCR a one-step procedure. The turn-around time of RT-PCR is 6 hours and for rtRT-PCR it is 2 hours.
  
  
  CONCLUSIONS
  For surveillance purposes nose and throat swabs are the more easy and practical to collect. It was proved that RT-PCR is too laborious, multi-step and time-consuming. The sensitivity of both assays is equal. The specificity of rtRT-PCR is higher. NAT assays for detection of influenza viruses have become an integral component of the surveillance programme in our country. They provide a fast, accurate and sensitive detection of influenza.
authors.affiliation: Institute of Public Health of the Republic of Macedonia, Skopje, R. Macedonia.
authors.forename: Golubinka
authors.initials: G
authors.lastname: Bosevska
authors.forename: Elizabeta
authors.initials: E
authors.lastname: Janceska
authors.forename: Gordana
authors.initials: G
authors.lastname: Kuzmanovska
authors.forename: Vladimir
authors.initials: V
authors.lastname: Mikik
authors.forename: Nikola
authors.initials: N
authors.lastname: Panovski
mesh_terms: D006801:Humans; D007251:Influenza, Human / Q000175:diagnosis*; D021141:Nucleic Acid Amplification Techniques; D011237:Predictive Value of Tests; D012367:RNA, Viral / Q000032:analysis; D060888:Real-Time Polymerase Chain Reaction / Q000379:methods*; D018843:Republic of North Macedonia; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods*; D012680:Sensitivity and Specificity; D013997:Time Factors
chemical_list: D012367:RNA, Viral