©The Author(s) 2015. Published by Baishideng Publishing Group Inc. All rights reserved.
World J Virology. Nov 12, 2015; 4(4): 365-371
Published online Nov 12, 2015. doi: 10.5501/wjv.v4.i4.365
Published online Nov 12, 2015. doi: 10.5501/wjv.v4.i4.365
Rapid genotyping of human rotavirus using SYBR green real-time reverse transcription-polymerase chain reaction with melting curve analysis
Yupin Tong, Department of Medicine, University of Alberta, Edmonton, Alberta T6G 2J2, Canada
Bonita E Lee, Department of Pediatrics, University of Alberta, Edmonton, Alberta T6G 2J2, Canada
Xiaoli L Pang, Department of Laboratory Medicine and Pathology, University of Alberta, Edmonton, Alberta T6G 2J2, Canada
Xiaoli L Pang, Provincial Laboratory for Public Health, University of Alberta Hospital, Edmonton, Alberta T6G 2J2, Canada
Author contributions: Tong Y performed the experiments, analyzed the data and wrote the manuscript; Lee BE contributed data and sample selections for the study and revised the manuscript; Pang XL designed the research and revised the manuscript.
Institutional review board statement: The study was reviewed and approved by the Provincial Laboratory for Public Health Edmonton Alberta (Provlab) Institutional Review Board.
Conflict-of-interest statement: None.
Data sharing statement: Technical appendix, clinical dataset is available from the corresponding author at xiao-li.pang@albertahealthservices.ca.
Correspondence to: Xiaoli L Pang, PhD, Provincial Laboratory for Public Health, University of Alberta Hospital, WMC 2B4.58, 8440 112 Street, Edmonton, Alberta T6G 2J2, Canada. xiao-li.pang@albertahealthservices.ca
Telephone: +1-780-4073483 Fax: +1-780-4078984
Received: May 29, 2015
Peer-review started: June 2, 2015
First decision: June 18, 2015
Revised: August 1, 2015
Accepted: September 29, 2015
Article in press: September 30, 2015
Published online: November 12, 2015
Processing time: 167 Days and 13.7 Hours
Peer-review started: June 2, 2015
First decision: June 18, 2015
Revised: August 1, 2015
Accepted: September 29, 2015
Article in press: September 30, 2015
Published online: November 12, 2015
Processing time: 167 Days and 13.7 Hours
Core Tip
Core tip: Genotyping rotavirus is essential for monitoring strain shifts in rotavirus surveillance and vaccine evaluation. Current conventional semi-nested real-time reverse transcription-polymerase chain reaction (RT-PCR), the most commonly used rotavirus genotyping assay is a labor-intensive, complex multi-step procedure and has long turn around-time. The newly developed SYBR Green real time RT-PCR assay is simple, fast and has comparable sensitivity and specificity as conventional semi-nested RT-PCR. This new assay was used to genotype clinical samples which tested positive for rotavirus from January 2012 to June 2013 and new emerging G12 strains were identified in Alberta, Canada.