title: Direct Identification of Enteroviruses in Cerebrospinal Fluid of Patients with Suspected Meningitis by Nested PCR Amplification.
abstract: Enteroviruses, the most common human viral pathogens worldwide, have been associated with serous meningitis, encephalitis, syndrome of acute flaccid paralysis, myocarditis and the onset of diabetes type 1. In the future, the rapid identification of the etiological agent would allow to adjust the therapy promptly and thereby improve the course of the disease and prognosis. We developed RT-nested PCR amplification of the genomic region coding viral structural protein VP1 for direct identification of enteroviruses in clinical specimens and compared it with the existing analogs. One-hundred-fifty-nine cerebrospinal fluids (CSF) from patients with suspected meningitis were studied. The amplification of VP1 genomic region using the new method was achieved for 86 (54.1%) patients compared with 75 (47.2%), 53 (33.3%) and 31 (19.5%) achieved with previously published methods. We identified 11 serotypes of the Enterovirus species B in 2012, including relatively rare echovirus 14 (E-14), E-15 and E-32, and eight serotypes of species B and 5 enteroviruses A71 (EV-A71) in 2013. The developed method can be useful for direct identification of enteroviruses in clinical material with the low virus loads such as CSF.
authors.affiliation: Belozersky Institute of Physico-Chemical Biology, Lomonosov Moscow State University, Moscow 119899, Russia. krasota@belozersky.msu.ru.|Chumakov Institute of Poliomyelitis and Viral Encephalitides, Moscow 142782, Russia. krasota@belozersky.msu.ru.
authors.forename: Alexandr
authors.initials: A
authors.lastname: Krasota
authors.affiliation: Hygienic and Epidemiological Center in the Omsk Region, Omsk 644116, Russia. polioom@mail.ru.
authors.forename: Natalia
authors.initials: N
authors.lastname: Loginovskih
authors.affiliation: Chumakov Institute of Poliomyelitis and Viral Encephalitides, Moscow 142782, Russia. poliom@aha.ru.
authors.forename: Olga
authors.initials: O
authors.lastname: Ivanova
authors.affiliation: Belozersky Institute of Physico-Chemical Biology, Lomonosov Moscow State University, Moscow 119899, Russia. galina.lipskaya@gmail.com.
authors.forename: Galina
authors.initials: G
authors.lastname: Lipskaya
mesh_terms: D000293:Adolescent; D000328:Adult; D002648:Child; D002675:Child, Preschool; D004770:Enterovirus / Q000145:classification / Q000235:genetics / Q000302:isolation & purification*; D004769:Enterovirus Infections / Q000134:cerebrospinal fluid / Q000175:diagnosis / Q000821:virology*; D005260:Female; D005838:Genotype; D006801:Humans; D008297:Male; D008581:Meningitis / Q000134:cerebrospinal fluid / Q000175:diagnosis / Q000821:virology*; D016133:Polymerase Chain Reaction / Q000379:methods*; D012189:Retrospective Studies; D014764:Viral Proteins / Q000235:genetics; D055815:Young Adult
chemical_list: D014764:Viral Proteins
keywords: Enterovirus; cerebrospinal fluids (CSF); detection; direct genotyping; identification; nested PCR; sequencing