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J Gen Virol 71 (1990), 2141-2147; DOI 10.1099/0022-1317-71-9-2141
© 1990 Society for General Microbiology

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Detection and differentiation of picornaviruses in clinical samples following genomic amplification

D. Michael Olive1,2,{dagger}, Siham Al-Mufti2, Wahiba Al-Mulla1, M. A. Khan3, Alexander Pasca1, Glyn Stanway3 and Widad Al-Nakib1,2,

1 Department of Microbiology, Faculty of Medicine, Kuwait University, P.O. Box 24923, 13110 Kuwait
2 Public Health Virology Laboratory, 5540 Shaab, 13056 Kuwait
and3 Department of Biology, University of Essex, Colchester CO4 3SQ, U.K.

A polymerase chain reaction (PCR) assay was used to detect and differentiate picornaviruses (PVs), using primers homologous to the 5' non-coding and VP2 regions of the PV genome. The PCR resulted in a 530 bp PCR product for human rhinoviruses (HRVs) and a 650 bp product for polioviruses, coxsackieviruses (CV) or echoviruses. The PCR assay could detect as little as 1 p.f.u. of virus in either cerebrospinal fluid (CSF) or stool, using ethidium bromide-stained gels. Standard strains of poliovirus, CV, echovirus and HRV were detected, with the exception of echovirus type 22. In contrast, heterologous viruses, such as herpes simplex virus, human cytomegalovirus, adenovirus, influenza virus and rotavirus, as well as human and monkey cell DNA, were not amplified. In nasal swabs taken from patients with respiratory infections, the PCR detected 27 of 28 HRV isolation-positive specimens. All specimens from which viruses other than HRVs were isolated were negative by PCR. The PCR definitively identified poliovirus and CVs from the CSF or stool of patients with aseptic meningitis, as well as CV in the pericardial fluid of a patient who had suffered a myocardial infarction. Specimens taken from patients with similar pathologies, and from which heterologous viruses were isolated, were uniformly negative by PCR.

{dagger} Present address: GENE-TRAK Systems, 31 New York Avenue, Framingham, Massachusetts 01701, U.S.A.

Received 20 March 1990; accepted 4 June 1990.


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