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 this study, we described a multiplex SYBR Green real-time PCR assay for detection of 12 respiratory RNA viruses, belonging to three families of viruses: Picornaviridae (RV and EV), Paramyxoviridae (RSV, HMPV, hPIV 1-4) and Coronaviridae (coronaviruses 229E, SARS, NL63, OC43). Enteroviruses and RVs can be detected in 25 -30% of respiratory tract infections. Nearly 80% of all children have experienced RV infections by the age of two years. Human EV type 68 has been identified in respiratory tract specimens and has been associated with respiratory diseases (14) . RSV is the most important cause of severe LRI in infants and young children worldwide. In temperate climates, RSV has been significantly documented as a winter epidemics agent of acute LRI (bronchiolitis and pneumonia) (2) . Some studies have reported difficulties in detection of RSV with swabs. RSV lability was suggested as a reason of decreasing the sensitivity of detection (15) . HMPV is the cause of 5 -7% of viral respiratory tract infections in hospitalized children (16) . HMPV outbreaks usually occur at the same time with the RSV season (17, 18) . According to the phylogenetic analysis, HMPV is the closest human virus to RSV (19) . (20) . Human respiratory coronaviruses infect all age groups. Nearly 10% of all URI and LRI in children are caused by these viruses (21) . Corona viruses are often co-detected with other respiratory viruses, especially RSV (22) , but in this study, we did not have co-infections. Respiratory disease can be caused by one of the known respiratory viruses. Nowadays, there are different methods for detection of viruses which may lead to different results. The use of some diagnostic methods such as cell culture might have long delays before the final results. Sensitive and rapid detection of viral infections is essential for reducing the nosocomial transmission and limiting the overuse of antibiotics (15) ; molecular detection methods have been widely used.