We describe here the results of respiratory syncytial virus (RSV) detection by reverse transcription polymerase chain reaction

Respiratory syncytial virus (RSV) is well recognized as the single most important pathogen accounting for acute viral infections of the lower respiratory tract in infants and young children. That is why rapid detection of RSV is mandatory for early diagnosis, isolation measures, and antiviral therapy. The aim of our stydy was to implement several tests for diagnosis of RSV and determination the role of this pathogen in high risk infants in Bulgaria. During the period 2003-2005 the Laboratory of Influenza and Acute Respiratory Diseases responded the new requirements for the diagnosis of RSV using a complex of classic and contemporary methods. HEp-2 cell lines were used for the isolation of the viruses -total of 7 RSV strains were isolated and identified by classical CFT. Rapid enzyme immunoassays -Directigen were used for the detection of RSV viral antigens of 16 passaged clinical samples. 12 positive by IFA modification on chamber slides were identified. In 2005 RT-PCR using N specific primers was applied for detection of RSV genome in initial samples from patients before viral isolation. The obtained 4 positive results by this method helped to decode the aetiology of the outbreak in Pazardjic in January 2005.Our work shows that the laboratory Influenza and Acute Rerspiratory Diseases have the readiness for effective diagnosis of RSV using a complex of contemporary methods for detection of RSV in clinical samples.

I In nt tr ro od du uc ct ti io on n/ /P Pu ur rp po os se e: : Reverse transcriptasepolymerase chain reaction (RT-PCR) for detection, typing, and subtyping of the hemagglutinin and neuraminidase genes of influenza type A (H3N2 and H1N1) viruses in patients' specimens has been developed and

Influenza viruses A and B are world-wide major causative agents of human acute respiratory infections. Three subtypes of HA (H1 to H3) and two subtypes of NA (N1 and N2) are found among influenza A viruses that have caused epidemics and pandemics among humans. For the rapid isolation and identification of influenza viruses in clinical samples variety of methods are available but the use of nucleic acid amplification techniques like RT-PCR has made sensitive diagnosis of influenza virus infection feasible, with the possibility of type and subtype determination. During the season 2002/2003 the laboratory of Influenza and ARD detected 7 type A and 4 type B influenza viruses by RT-PCR in 15 simulated samples from the respiratory panel from SEQC. In the season 2003 /2004 -42 positive results for influenza viruses subtype А/Н3N2/ were obtained by RT-PCR using specific НA and NA primer pairs from 50 nasal and throat swabs from patients with influenza-like diseases. The RT-PCR provides a sensitive and specific method for detecting, typing and subtyping influenza viruses A and B in clinical samples and can be used in the routine diagnosis of the disease in Bulgaria together with a large complex of classical methods.

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