Development of a real-time RT-PCR and Reverse Line probe Hybridisation assay for the routine detection and genotyping of Noroviruses in Ireland.
BACKGROUND
Noroviruses are the most common cause of non-bacterial gastroenteritis. Improved detection methods have seen a large increase in the number of human NoV genotypes in the last ten years. The objective of this study was to develop a fast method to detect, quantify and genotype positive NoV samples from Irish hospitals.


RESULTS
A real-time RT-PCR assay and a Reverse Line Blot Hybridisation assay were developed based on the ORF1-ORF2 region. The sensitivity and reactivity of the two assays used was validated using a reference stool panel containing 14 NoV genotypes. The assays were then used to investigate two outbreaks of gastroenteritis in two Irish hospitals. 56 samples were screened for NoV using a real-time RT-PCR assay and 26 samples were found to be positive. Genotyping of these positive samples found that all positives belonged to the GII/4 variant of NoV.


CONCLUSIONS
The combination of the Real-time assay and the reverse line blot hybridisation assay provided a fast and accurate method to investigate a NoV associated outbreak. It was concluded that the predominant genotype circulating in these Irish hospitals was GII/4 which has been associated with the majority of NoV outbreaks worldwide. The assays developed in this study are useful tools for investigating NoV infection.
Menton
John F
JF
Lab 439, Food Science Building, Department of Microbiology, University College Cork, Cork, Republic of Ireland. j.menton@ucc.ie
Kearney
Karen
K
Morgan
John G
JG
D017250:Caliciviridae Infections / Q000453:epidemiology / Q000821:virology*; D004196:Disease Outbreaks; D005759:Gastroenteritis / Q000453:epidemiology / Q000821:virology*; D005838:Genotype; D006801:Humans; D007494:Ireland / Q000453:epidemiology; D029322:Norovirus / Q000145:classification* / Q000235:genetics / Q000302:isolation & purification*; D009693:Nucleic Acid Hybridization / Q000379:methods*; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods*; D012680:Sensitivity and Specificity