NESTED PCR FOR SPECIFIC DETECTION AND RAPID IDENTIFICATION OF HUMAN PICORNAVIRUSES

A nested PCR for the detection and rapid identification of human picor naviruses is described. Enteroviruses and rhinoviruses were amplified with the same set of four primers from the 5'-noncoding region. The ne sted primers allowed the detection of far less than 1 PFU in diluted v irus stocks without Southern blot hybridization. In patients with neur ological disorders (mainly aseptic meningitis), 43% of 37 specimens (1 1 of 21 cerebrospinal fluid specimens, 2 of 10 serum specimens, and 3 of 6 stool specimens) were positive by PCR. A total of 21% (10 of 47 s pecimens) of heart biopsy specimens from patients with dilative cardio myopathy were PCR positive, whereas 3% (2 of 70 specimens) of control biopsy specimens from patients with coronary artery disease were PCR p ositive. PCR-amplified fragments from 27 of 29 clinical isolates and 1 4 of 28 patient samples were successfully serotyped by restriction enz yme digestion. Two specimens were further investigated by direct seque ncing of PCR products, leading to the identification of a poliovirus t ype 3 isolate with a sequence that was highly divergent from previousl y published sequences.