Andrea Maloney presents her defense on Monday, November 28th at 3:00pm

(Under the direction of Dr. Jan Vinji)

Noroviruses (NoVs) are now recognized as the leading acute of outbreaks and cases of nonbacterial, acute gastroenteritis in humans. With its low infectious dose, NoVs have been implicated in numerous outbreaks caused by exposure to contaminated water, which raises the need for rapid, sensitive and accurate methods to monitor for NoV in environmental water samples. In this study, we compared the detection limit of 13 (7 GI, 6 GII) different primer combinations targeting the 5′-end of ORF2 (region C) by conventional RT-PCR followed by hemi-nested PCR. A panel of 24 norovirus strains four different GI genotypes (GI.1, GI.2, GI.3, GI.4) and seven different GII genotypes (GII.1 through GII.7) was used for these evaluations. The primer combinations COG1F/G1SKR and G1SKF/G1SKR for GI and COG2F/G2SKR and G2SKF/G2SKR for GII NoV showed the lowest detection limit across the range of genotypes tested. We compared these hemi-nested PCR assays with a recently developed TaqMan.-based real-time RT-PCR assay on naturally contaminated water samples collected between Sept 2003 -Sept 2004 at drinking water and waste water utilities in Georgia. None of the drinking water samples and 63% of the waste water samples tested positive for NoVs. While hemi-nested PCR is as sensitive as real-time PCR detection and is capable to compare clinical and environmental strains by sequence analysis of the PCR products, TaqMan. real-time RT-PCR is a superior method for rapid and simple monitoring of environmental waters for NoVs.

Committee Members:
Dr. Jan Vinji
Dr. Louise Ball
Dr. Mark Sobsey

For further information please contact Rebecca Riggsbee Lloyd by email at Rebecca_Lloyd@unc.edu