Figure 3

Scheme for the detection, identification, and genotyping of enteric viruses. A pattern of positive zipcode signals are used to detect and identify viruses as well as determine the genotypes of noroviruses and rotaviruses. Representative patterns of zipcode signals from 6 samples and their interpretation are shown. The normalized signal to background fluorescent signal (S/B) for each zipcode target for a given sample is indicated. An S/B value >3 fold higher than the median signal of negative controls for a given zipcode target was defined as a positive signal. The assay has built-in redundancy with at least 3 zipcode addresses assigned to identify each virus. Only 2 (for calicviruses) or 3 (for rotaviruses) positive signals are required for the identification of a virus or genotype. For example, sample 3 was identified as norovirus GGII based on 2 positive zipcode signals (9 and 12) out of the 4 designated zipcode addresses for norovirus (9, 10, 11 and 12). Similarly, samples 5 and 6 were determined as rotavirus P[6] based on a unique zipcode signature of 2 out of 3 zipcode addresses assigned (See Supplementary Fig. 1 for details of genotype determination of rotavirus). ^†Designated zipcodes did not provide positive signals as indicated in red; however, identification was achieved successfully since the criteria of 2 out of 4 positive zipcodes was met.