Detection of enteroviral RNA by a new single-step PCR

Abstract

In this study we present a modified diagnostic routine PCR (polymerase chain reaction) technique for the detection of specific enteroviral nucleic acid sequences in human body fluids, the use of which would reduce the number of false-positive results, the costs and the concentration of reagents required in PCR. Cerebrospinal samples, pharyngeal swabs and faeces were tested. To this end, general primers, selected in the highly conserved 5′ non-coding region were used, with a closed-tube RT semi-nested PCR protocol, and the PCR product was analysed using a hybridization in solid phase. A total of 32 patients with suspected enteroviral infection, 17 with suspected viral meningitis and 15 with a possible acute enteroviral infection different from meningitis were analysed, and 80% of the patients with possible acute enteroviral infection were PCR-positive. A broad range of enteroviruses can be detected and false-positive results can be reduced. The availability of results in less than 12 h and the lack of need for radiolabelled probes also increase the convenience of this protocol.

Key Words:

• enterovirus
• PCR