Evaluation of the sensitivity and specificity of p

Authors: Cheonghoon Lee Sooryun Cheong HeeJung Lee Miye Kwon Ilnam Kang EunGyoung Oh HongSik Yu SoonBum Shin SangJong Kim

Publish Date: 2010/11/03

Volume: 48, Issue: 5, Pages: 586-593

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Abstract

Noroviruses NoV are the key cause of acute epidemic gastroenteritis and oysters harvested from NoVpolluted sea areas are considered as the significant vectors of viral transmission To improve NoV detection from oyster using nested reverse transcriptionpolymerase chain reaction RTPCR we evaluated the sensitivity and specificity of previously published primer pairs and the efficiency of different RNA extraction procedures Among the primer pairs used for RTPCR the sensitivity of GIF1/GIR1GIF2/GIR1 and GIIF1/GIIR1GIIF2/GIIR1 was higher than that of other primer pairs used in nested RTPCR for the detection of NoV genogroup I NoV GI and NoV GII from both NoVpositive stool suspension and NoVseeded oyster concentrates respectively the resulting products showed neither unspecific bands in the positive samples nor falsepositive bands in the negative controls The extraction of NoV RNA from oyster samples using a QIAamp® Viral RNA Mini kit with a QIAshredder™ Homogenizer pretreatment afforded more efficient recovery mean recovery for NoV GI and GII 64 and the procedure was less time consuming 30 min than most other RNA extraction procedures The results of RNA extraction procedure and primer pairs evaluated by nested RTPCR assay in this study can be useful for monitoring NoV contamination in oysters which is an indicator of possible public health risks

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