Detection of enteroviruses using subgenomic probes of coxsackie virus B4 by hybridization

The objective of this research was to develop group- and type-specific probes for the detection of enteroviruses. Coxsackie virus B4 RNA was cloned, and a series of subgenomic clones were generated. Six of these clones, containing sequences from the 3′ end or the 5′ end of the genome, were tested for their ability to detect these viruses in a small number of infected cells employing nucleic acid hybridization technique and total cytoplasmic RNA from a panel of 11 serotypes of enteroviruses. The RNA from cells infected with Coxsackie B viruses gave characteristic and positive hybridization signals. Coxsackie B-specific probes and a control Echo 9 probe detected Coxsackie A9 and Echo 3 weakly. As little as 0.5 μg of the RNA—which contained 10–20 ng of poly(A)-containing, virus-specific, hybridizable RNA—was sufficient to successfully conduct the assay, suggesting high sensitivity of these probes. Probes that are 3′ end-specific appear to be group specific, while those that are 5′ end-specific appear to be type specific among the serotypes tested.