A fatal case of enterovirus 71 infection with pulmonary edema and rhombencephalitis occurred in Brest, France, in April 2007. The virus was identified as subgenogroup C2. This highly neurotropic enterovirus merits specific surveillance outside the Asia-Pacific region.

NS3 protease is essential for hepatitis C Virus (HCV) replication, and is one of the most promising targets for specific anti-HCV therapy. Its natural polymorphism has not been studied at the quasispecies level. In the present work, the genetic heterogeneity of the NS3 protease gene was analyzed in 17 HCV genotype 1 (5 subtypes 1a and 12 subtypes 1b) samples collected from infected patients before anti-viral therapy. A total of 294 clones were sequenced. Although the protease NS3 is considered to be one of the less variable genes in the HCV genome, variability of both nucleotide and amino acid sequences was found. In variants belonging to 1a and 1b subtypes, 224 and 267 of 543 positions showed one or more nucleotide substitutions, respectively. Forty and 74 of the 181 NS3 amino acid positions showed at least one mutation in HCV-1a and HCV-1b isolates, respectively. Most substitutions were conservative. This substantial polymorphism of the NS3 protease produced by HCV-1a and HCV-1b suggests that, despite the numerous functional and structural constraints, the enzyme is sufficiently flexible to tolerate substitutions.

Thirteen laboratories participated in blind tests of a panel of 20 coded cerebrospinal fluid specimens (7 uninfected samples, 3 samples infected with 1 50% tissue culture infective dose [TCID 50 ]/0.1 ml [nonenterovirus strains], and 10 samples infected with 10, 1, or 0.1 TCID 50 /0.1 ml [three different enterovirus serotypes]) on the Amplicor enterovirus PCR assay (Roche Diagnostic Systems). The panel was also evaluated by in-house PCR (two nested-PCR and three one-step PCR assays) or tissue culture (eight laboratories). The viral load was shown to influence 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 TCID 50 /0.1 ml, respectively; titers of 0.1 TCID 50 /0.1 ml resulted in a sensitivity of only 16%. The overall specificity of the Amplicor test was 98%. The Amplicor assay compared favorably to the five in-house PCR tests (no significant difference in either sensitivity or specificity) and was much more sensitive than tissue culture (P < 0.001), even for high viral loads. It was easy to perform, rapid (about 6 h), well-standardized, and appeared to be suitable for the diagnosis of enterovirus meningitis on a routine basis in laboratories trained in molecular biology techniques.

Four consecutive epidemics of keratoconjunctivitis caused by adenovirus 8 (Ad8) occurred over a 5-year period in Brest, France. A selection of 30 strains isolated during this period was studied by DNA restriction enzyme analysis using nine restriction enzymes. BglI and SacI were the most discriminative enzymes and allowed the recognition of four DNA variants, all different from the prototype strain Trim. Within each of the epidemics, the strains tested could not be distinguished in this analysis. Between strains from different epidemics differences in DNA structure could be detected however. Thus, the Ad8 epidemics of 1983/1984, 1984, 1987, and 1988 appear to have been due to DNA variants Ad8/D7, D8, D9, and D10, respectively. These results demonstrate that the DNA of Ad8 seems to display a considerable variability, comparable to that observed with Ad7 and Ad21. As has been described for Ad7, Ad21 and Ad41, successive DNA variants of Ad8 prevail during one or more years, and are then replaced by other, newly emerging variants sometimes associated with epidemics.

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