NONISOTOPIC AUTOMATABLE MOLECULAR PROCEDURES FOR THE DETECTION OF ENTEROVIRUSES

Five microwell non isotopic hybridization assays, based on colorimetri c immunoenzymatic reading, were developed and evaluated for the rapid and automatable detection of enteroviruses. Virus nucleic acids and/or capture probes were covalently bound to microtiter wells, and digoxig enin-11-dUTP was used as label for the detection of hybridized materia l. Among these procedures, a reverse transcriptase polymerase chain re action (RT-PCR) hybridization assay was the most sensitive, enabling t he detection of 10 MPNCU of poliovirus, and offering detection specifi city for other enteroviruses, such as coxsackieviruses and echoviruses . The second most sensitive method was a complementary hybridization a ssay, simultaneously using three detection probes, one from the 5' end and two from the 3' end of poliovirus genome, offering a sensitivity for poliovirus detection of 5 x 10(3) MPNCU. (C) 1996 Academic Press L imited