Key result
In Situ Capture RT-qPCR (ISC-RT-qPCR) demonstrated a 10- to 100-fold greater sensitivity than traditional RT-qPCR for detecting human noroviruses and significantly increased the detection rate of GI HuNoVs in retail oyster gills (33.3% vs 5.6%).
ISC-RT-qPCR is a more sensitive and simpler method than traditional RT-qPCR for detecting Human Noroviruses in clinical and oyster samples.
May enhance HuNoV detection in oysters; leaves open whether it improves food safety surveillance or reduces outbreaks.
capture RT-qPCR (ISC-RT-qPCR) to detect HuNoVs in oysters and compared with the traditional RT-qPCR method. Ten HuNoVs RT-PCR positive and 5 negative clinical samples from gastroenteritis patients were used to compare specificity and sensitivity of ISC-RT-qPCR against that of the RT-qPCR assay. ISC-RT-qPCR had at a one-log and a two-log increase in sensitivity over that of the RT-qPCR assay for genotype I (GI) and GII, respectively. Distributions of HuNoVs in oyster tissues were investigated in artificially inoculated oysters. GI HuNoVs could be detected in all tissues in inoculated oysters by both ISC-RT-qPCR and RT-qPCR. GII HuNoVs could only be detected in gills and digestive glands by both methods. The number of viral genomic copies (vgc) measured by ISC-RT-qPCR was comparable with RT-qPCR in the detection of GI and GII HuNoVs in inoculated oysters. Thirty-six oyster samples from local market were assayed for HuNoVs by both assays. More HuNoVs could be detected by ISC-RT-qPCR in retail oysters. The detection rates of GI HuNoVs in gills, digestive glands, and residual tissues were 33.3, 25.0, and 19.4% by ISC-RT-qPCR; and 5.6, 11.1, and 11.1% by RT-qPCR, respectively. The detection rates of GII HuNoVs in gills were 2.8% by ISC-RT-qPCR; no GII HuNoV was detected in these oysters by RT-qPCR. Overall, all results demonstrated that ISC-RT-qPCR is a promising method for detecting HuNoVs in oyster samples.
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Zhou et al. (2017) studied Human norovirus contamination. In Situ Capture RT-qPCR (ISC-RT-qPCR) vs. RT-qPCR was evaluated on Detection rate of GI HuNoVs in gills of retail oysters (p=<0.05). In Situ Capture RT-qPCR (ISC-RT-qPCR) demonstrated a 10- to 100-fold greater sensitivity than traditional RT-qPCR for detecting human noroviruses and significantly increased the detection rate of GI HuNoVs in retail oyster gills (33.3% vs 5.6%).
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