Analytical performance of norovirus real-time RT-PCR detection protocols in Canadian laboratories

作者：Mattison Kirsten^*; Grudeski Elsie; Auk Brian; Brassard Julie; Charest Hugues; Dust Kerry; Gubbay Jonathan; Hatchette Todd F; Houde Alain; Jean Julie; Jones Tineke; Lee Bonita E; Mamiya Hiroshi; McDonald Ryan; Mykytczuk Oksana; Pang Xiaoli; Petrich Astrid; Plante Daniel; Ritchie Gordon; Wong Julie; Booth Tim F

来源：Journal of Clinical Virology, 2011, 50(2): 109-113.

DOI：10.1016/j.jcv.2010.10.008

• Norovirus
• Real-time reverse transcription PCR
• Method comparison

Background: Noroviruses (NoVs) are the leading cause of infectious gastroenteritis worldwide. Real-time reverse transcription PCR (real-time RT-PCR) is the preferred method of NoV detection for the majority of testing laboratories. Although the accepted target region for molecular detection assays is the conserved ORF1/ORF2 junction, multiple variations have been published with differences in primers, probes, reagents, multiplexing, etc.
Objectives: We assessed the detection limit for GII.4 NoV real-time RT-PCR assays as well as the ability to detect the non-GII.4 NoV genotypes in each participating laboratory.
Study design: A panel of 25 RNA samples was circulated to 18 testing laboratories for comparison of their real-time RT-PCR procedures for NoV detection.
Results: Multiple protocols with slight differences in reagents or conditions successfully detected 10 genome equivalents or fewer of NoV per reaction. Multiplex procedures were significantly associated (p = 0.04) with false negative results, particularly for a GI.2 strain. Sensitive detection was associated with false positive results (p = 0.03).
Conclusions: Overall, the data indicate that comparable results are produced under slightly different assay conditions.

• 出版日期2011-2
• 单位Saskatchewan