Development and validation of the real-time PCR assay for the identification of viral agents causing acute respiratory infections in humans

• Published in: Molekulârnaâ genetika, mikrobiologiâ i virusologiâ no. 4; p. 32
• Main Authors: Sergeeva, E I, Ternovoĭ, V A, Demina, O K, Demina, A V, Korneev, A N, Shikov, A N, Berillo, S A, Agafonov, A P, Sergeev, A N
• Format: Journal Article
• Language: Russian
• Published: Russia (Federation) 2013
• Subjects: Animals; Birds - virology; Humans; Influenza A Virus, H1N1 Subtype - genetics; Influenza A Virus, H1N1 Subtype - isolation & purification; Influenza A Virus, H3N2 Subtype - genetics; Influenza A Virus, H3N2 Subtype - isolation & purification; Influenza A Virus, H5N1 Subtype - genetics; Influenza A Virus, H5N1 Subtype - isolation & purification; Influenza B virus - genetics; Influenza B virus - isolation & purification; Influenza in Birds - diagnosis; Influenza in Birds - virology; Influenza, Human - diagnosis; Influenza, Human - virology; Real-Time Polymerase Chain Reaction - methods; Respiratory Tract Infections - diagnosis; Respiratory Tract Infections - genetics; Respiratory Tract Infections - virology
• ISSN: 0208-0613

The real time PCR assay targeting influenza A and B virus, 5 subtypes of influenza A virus (seasonal H1N1, pandemic H1N1 (2009), seasonal H3N2, pathogenic for human subtypes of avian influenza H5 and H7), respiratory syncytial virus, and adenovirus was developed. The analytical sensitivity of the developed assay was 1 x 10(3) genome equivalents per ml. The diagnostic sensitivity of the method was 1 x l0(3)-10(4) viral particles per ml. Experiments with human DNA/cDNA and viral cDNA showed a markedly high diagnostic specificity of the developed PCR assay. In the assay of the developed PCR test, 50 nasopharyngeal swab specimens were tested. The etiology was identified in 33 samples.