MULTICENTER EVALUATION OF A COMMERCIALLY AVAILABLE PCR ASSAY FOR DIAGNOSING ENTEROVIRUS INFECTION IN A PANEL OF CEREBROSPINAL-FLUID SPECIMENS

Thirteen laboratories participated in blind tests of a panel of 20 cod ed cerebrospinal fluid specimens (7 uninfected samples, 3 samples infe cted with 1 50% tissue culture infective dose [TCID50]/0.1 ml [nonente rovirus strains], and 10 samples infected with 10, 1, or 0.1 TCID50/0. 1 ml [three different enterovirus serotypes]) on the Amplicor enterovi rus PCR assay (Roche Diagnostic Systems). The panel was also evaluated by in-house PCR (two nested-PCR and three one-step PCR assays) or tis sue culture (eight laboratories). The viral load was shown to influenc e greatly the sensitivity of the assay. The average sensitivity of the Amplicor test ranged from 67 to 98% for viral titers of 1 to 10 TCID5 0/0.1 ml, respectively; titers of 0.1 TCID50/0.1 ml resulted in a sens itivity of only 16%. The overall specificity of the Amplicor test was 98%. The Amplicor assay compared favorably to the five in-house PCR te sts (no significant difference in either sensitivity or specificity) a nd was much more sensitive than tissue culture (P < 0.001), even for h igh viral loads. It was easy to perform, rapid (about 6 h), well-stand ardized and appeared to be suitable far the diagnosis of enterovirus m eningitis on a routine basis in laboratories trained in molecular biol ogy techniques.