FIG. 1.

Principle and steps of the microarray procedure for genotyping of HRV. Sets of capture oligonucleotides (oligos) specific for the G and P types of HRV are covalently immobilized on glass microscope slides (A). With these oligonucleotides as probes, the RT-PCR products containing type-specific regions of the VP7 and VP4 genes are captured on the microarrays by hybridization under low-stringency conditions (B). Oligonucleotides with 3′ ends that are matched to the sequences of the captured templates are extended with a mixture of deoxynucleoside triphosphates containing cyanine 5 (Cy5)-labeled dUTP by using a thermostable DNA polymerase (C). The fluorescence incorporated in the sequence-specific primer extension reaction is measured in a microarray scanner (D). The results are interpreted by visual inspection of the arrays or by calculation of the relative fluorescence intensity of the individual spots.