Application Guide - QIAxcel Advanced

66 QIAxcel Advanced Application Guide 10/2016 A Novel Approach for Identification of 16 Respiratory Viral Targets Xuejun Ma, Chen Zhang Key Laboratory for Medical Virology, Ministry of Health, National Institute for Viral Disease Control and Prevention, Chinese Center for Disease Control and Prevention, No. 155, Changbai Road, Changping District, Beijing 102206, China Introduction Viral respiratory tract infections are associated with various types of virus, including adenovirus, coronavirus, human rhinovirus, influenza A, influenza B, metapneumovirus, parainfluenza and respiratory syncytial virus. To efficiently identify all these viruses requires the establishment of an accurate and reliable method that is rapid, sensitive and affordable. Current non-molecular methods, such as viral culture or immunofluorescence (direct fluorescence antibody, DFA), are insufficiently sensitive, time consuming and labor intensive (1,2). Consequently, molecular methods are used and a well-accepted approach is an automated electrophoresis system to identify differences in amplicon size. Recently, multiplex RT-PCR assays have been developed specifically to detect respiratory viruses, but the limited availability of required equipment renders them difficult to standardize. The method described herein uses a two-tube multiplex reverse-transcription PCR assay followed by amplicon-size separation using the QIAxcel® Advanced System (3). It is fast, highly automated, and based on reliable, affordable and readily available equipment and reagents. As such, this method may make an important contribution to routine virus identification. Materials and Methods Samples were collected and total RNA/DNA was extracted using the QIAamp® Viral RNA Mini Kit. A two-tube multiplex reverse-transcription PCR assay (two-tube assay) was used to detect 16 respiratory viruses based on their amplicon size differences.

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