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

During the September 2012 through September 2013, 3276 throat swab specimens were collected from hospitalized patients of all ages with symptoms of ILI. Initially, all the specimens were tested for influenza viruses by superscript III platinum one-step quantitative real-time PCR assay (Invitrogen, USA), selective for the matrix gene; 2514 samples were negative for influenza viruses. Of which 172 samples were randomly selected for this study. Of 172 specimens, the male:female proportion was 96:76. The mean age was 33.63 years and the median was 31 years. Of 172 samples, in the first set we did not detect any RSV, RV, EV or HMPV. In the second set, there was no PIV1 and PIV2, but 19 (11.04%) PIV3 and 9 (5.23%) PIV4 were detected. In the set of coronaviruses NL63, 229E, SARS, OC43, the only detected virus was 1 (0.58%) NL63. This was the first time that human corona virus NL63 was detected in Iran. All the PIV3-and PIV4-positive samples were detected during December to March ( Figure 1 ) and one corona virus NL63 belonged to a 28-day-old newborn in February. Among positive samples, the male:female proportion was 20:9, indicating that males were more affected than females. The mean age was 33.27 years and the median was 32 years. The standard curve and melt curves for positive samples are shown in Figures 2 and 3.