Multiplex SYBR Green Real-Time PCR Assay for Detection of Respiratory Viruses

Sultani, Mozhdeh; Mokhtari Azad, Talat; Eshragian, Mohammadreza; Shadab, Azadeh; Naseri, Maryam; Eilami, Owrang; Yavarian, Jila

In some particular situations, the emergence of respiratory viruses has an important impact on health systems worldwide, as happened with the emergence of severe acute respiratory syndrome (SARS)-coronavirus in 2003 which needed emergency diagnosis (8) . The development of molecular methods for direct detection of viruses has recently been progressed (9) . Polymerase chain reaction (PCR) has become an accepted tool in research since 1984 by Kary Mullis et al. (10) and is being introduced into laboratories (11). Singleplex PCR assays amplify one target in different reactions; they are resource intensive and expensive. Multiplex PCR uses a mixture of several distinctive primer pairs in one amplification reaction at the same time (12) . Nowadays, various multiplex reverse transcription assays have been developed (13) , but until now, there has not been enough study on SYBR Green multiplex real-time PCR for detection of respiratory viruses, a technique shown to be fast and cost effective.