multiplex sybr green real-time pcr assay for detection of respiratory viruses

conclusions multiplex sybr green real-time pcr is a rapid and relatively inexpensive method for detection of respiratory viruses. results application of the multiplex sybr green real-time pcr in clinical samples from 172 patients in a one-year study resulted in detection of 19 (11.04%) piv3, 9 (5.23%) piv4, and 1 (0.58%) coronavirus nl63. all the positive samples were detected during december to march (2011 - 2012). materials and methods randomized 172 influenza-negative respiratory specimens of all age groups of hospitalized patients were collected. after rna extraction, cdna was synthesized. three sybr green multiplex real-time pcr assays were developed for simultaneous detection of 12 respiratory rna viruses. each set of multiplex methods detected four viruses, the first set: respiratory syncytial virus, human metapneumovirus, rhinovirus, enterovirus; the second set: parainfluenza viruses 1 - 4 (piv1-4); the third set: coronaviruses nl63, 229e, severe acute respiratory syndrome (sars), and oc43. objectives the objective of this study was recognizing the panel of respiratory rna viruses by multiplex sybr green real-time polymerase chain reaction (pcr). background it is often difficult for a physician to distinguish between viral and bacterial causes of respiratory infections and this may result in overuse of antibiotics. in many cases of community-acquired respiratory infections, clinicians treat patients empirically. the development of molecular methods for direct detection of viruses has been progressed recently.