Prospective identification of enteroviruses involved in meningitis in 2006 through direct genotyping in cerebrospinal fluid

CHU Clermont-Ferrand, Laboratoire de Virologie, Centre de Biologie, F-63003 Clermont-Ferrand, France; Université d'Auvergne, Laboratoire de Virologie, EA3843, Faculté de Médecine, F-63001 Clermont-Ferrand, France

^* To whom correspondence should be addressed. Email: amirand{at}chu-clermontferrand.fr.

	Abstract

Enterovirus infections were investigated with special emphasis on performing rapid molecular identification of enterovirus serotypes responsible for aseptic meningitis directly in cerebrospinal fluid (CSF). Enterovirus genotyping was carried out directly with specimens tested for the diagnostic procedure, using two semi-nested PCR assays designed for amplifying the complete and partial gene sequences encoding the VP1 and VP4/VP2 capsid proteins, respectively. The method was used for identifying the enterovirus serotypes involved in meningitis in 45 patients admitted in 2005. Enterovirus genotyping was achieved in 98% of the patients studied and evidenced 10 of the most frequent serotypes identified earlier by genotyping of virus isolates. The method was applied for the prospective investigation of 54 patients with meningitis admitted consecutively in 2006. The enterovirus serotypes involved were identified with the CSF of 52 patients (96%) and comprised 13 serotypes within the HEV-B species and 1 within the HEV-A species. The three most frequent serotypes were echovirus 13 (E13; 24%), E6 (23%) and coxsackievirus B5 (11.5%), a pattern different from that observed in 2005. Genotyping of virus isolates was also performed in 35 patients in 2006 (meningitis, n=31; other diseases, n=4). By comparison, direct genotyping in CSF yielded a more complete pattern of enterovirus serotypes, thereby allowing the detection of rare serotypes: three less common serotypes (CB2, E21 and E27) were not detected by indirect genotyping alone. The study shows the feasibility of prospective enterovirus genotyping within one week in a laboratory setting.