Development of a sensitive real-time PCR for simultaneous detection and subtyping of influenza A and B viruses


Abstract


A new real-time PCR assay, using melting curve analysis, was developed for the rapid and reliable detection and sub-typing of influenza A and B.

In order to evaluate it’s specificity, cell culture surnatants positive for Respiratory Syncytial Virus, Parainfluenza Viruses 1, 2 and 3, Measles Virus, Influenza A (to evaluate Influenza B primer) and B (to evaluate Influenza A primer) were tested and all of the results were negative.

A series of Influenza A and B cell culture-grown viruses were diluted in virus transport medium, titrated and tested to determine the analytical sensibility which equated to 0.64, 0.026, 0.64, 0.62 PFU for A/H1N1, A/H3N2, Victoria-like and Yamagata-like B viruses, respectively. Twenty-five specimens, collected during the 2001/02 and 2002/03 seasons, which were positive for A/H1N1 (n = 7), A/H3N2 (n = 10), B Victoria-lineage (n = 5) and B Yamagata-lineage (n = 3), were tested in order to evaluate the assay’s clinical sensitivity, all of the results were positive.

The new real-time PCR appears to be a suitable tool for virological surveillance and the diagnosis of respiratory infections.


Keywords


Influenza viruses; real-time; virological surveillance

Full Text:

PDF


DOI: https://doi.org/10.2427/5997

NBN: http://nbn.depositolegale.it/urn%3Anbn%3Ait%3Aprex-8757

References



Refbacks

  • There are currently no refbacks.
We use cookies to ensure that we give you the best experience on our website. If you continue to use this site we will assume that you are happy with it (Read more).
Ok