A new Multiplex reverse transcription polymerase chain reaction assay for detection of influenza and respiratory syncytial virus

Between January 2002 and March 2004, oro-pharyngeal swabs taken from 164 subjects with acute respiratory symptoms were analysed. Specimens were examined using the Multiplex reverse transcription-polymerase chain reaction (RT-PCR) and tested with the specific primers of the following respiratory viruses: RSVNP-5B1, RSVNP-3B1, RSVNP-5E1, RSVNP-3E1, NP-5B1, NP-3B1, HA1-5E1, HA1-3E1, HA3-5A1, HA3-3A1, for respiratory syncytial virus and influenza viruses, respectively. The sampling periods considered were distributed as follows: January 2002/April 2002 (50 samples), December 2002/April 2003 (64 samples), February/March 2004 (50 samples). Of these samples, all of which taken from subjects who at the time of sampling presented fever (38.8°C), coughing and arthralgia, 11 resulted positive. Of these, 1 resulted positive for type B influenza virus, 8 for type A/H3N2 virus and 2 for RSV. Multiplex RT-PCR, compared favourably with the classic single-template polymerase chain reaction (PCR simplex), thus confirming this technique to be a valid, rapid and efficient tool, since it co-amplifies different target RNAs, at the same time identifying different types of virus.