[The prevalence of antibodies against Hepatitis E Virus in the Czech Republic: serological survey]. / Prevalence protilátek proti viru hepatitidy E v Ceské republice - sérologický prehled.

AIM: To determine the prevalence of antibodies against hepatitis E virus in the general population of the Czech Republic of age 15 to 64, to analyse the age and sex distribution of these antibodies, and to evaluate the benefit of the immunoblot test for the confirmation of the specificity of the enzyme immunoassay (EIA) screening test. MATERIAL AND METHODS: Sera from the last available multipurpose serological survey conducted in 2001 were tested. Anti-HEV IgG was detected by the RecomWell HEV IgG EIA test (Mikrogen Diagnostik, Germany). The immunoblot assay RecomLine HEV IgG/IgM (Mikrogen Diagnostik, Germany) was used for confirmation. RESULTS: Using the RecomWell IgG EIA test, anti-HEV IgG reactivity was found in 115 (6.7%) of 1715 sera. No significant difference in the anti-HEV IgG reactivity was found between men 58 (6.9%) and women 57 (6.6%). The prevalence of anti-HEV IgG increased with age from 3.5% in the age group 15-24 years to 16.8% in 55-64-year-olds. CONCLUSIONS: The prevalence of hepatitis E IgG antibodies determined in the serological survey in the age group 15-64 years was 6.7%. Recalculated for the general population of the Czech Republic, the prevalence was 8.6%. The prevalence of anti-HEV antibodies increased with age, reaching a peak of 16.8% in the age group 55-64 years. The prevalence was not significantly different between men and women. Using the immunoblot RecomLine IgG test for the confirmation of the specificity of the screening test in the seroprevalence study was not of clear benefit.

[Enterovirus sequencing as a new approach to the laboratory diagnosis for clinical and epidemiological purposes]. / Sekvenace enteroviru jako nový prístup k laboratorní diagnostice pro klinické i epidemiologické úcely.

OBJECTIVE: Introducing enterovirus sequencing as an advanced approach to classify the viruses isolated according to the novel nomenclature and to characterize isolates in detail. MATERIAL AND METHODS: Seventy-five specimens collected from 64 patients in two hospitals, Liberec Regional Hospital, and Plzen University Hospital, were analyzed. The study patients' age ranged from four to 54 years, with a median of 15 years in males and 16 years in females. In most patients, the reasons for admission were intense headache, fever, vomiting, tiredness, meningeal symptoms, intestinal symptoms (in two patients), and skin symptoms (in one patient). The specimens collected were rectal and throat swabs, cerebrospinal fluid (CSF) and stool specimens. Molecular detection and typing were performed using the RT-PCR method. A segment of the 5´non-coding RNA was selected for typing. Specimens were amplified using single-step PCR with external primers and with the same primers extended to include M13 sequences (Generi-Biotech). The LASERGENE software (DIASTAR) was used in sequence editing, alignment, and quality check. The sequences obtained were checked against the central GenBank sequence database using the BLAST algorithm. RESULTS: The identification of the study isolates resulted in 61 ECHO viruses 30, three coxsackie viruses B1, one coxsackie virus B3, one coxsackie virus A9, one enterovirus 86, one enterovirus 71, Two ECHO viruses 13/coxsackie virus B5, one ECHO virus 7/30/coxsackie virus B4, one coxsackie virus B4/enterovirus B, one enterovirus 87/ECHO virus 30/enterovirus B, and one ECHO virus 3. All viruses isolated, except enterovirus 71 classified into group A, were of group B. CONCLUSION: The enteroviruses were identified unambigously, although the sequencing only targeted a short, conserved segment that showed considerable variability. The sequencing was an effective alternative to enterovirus identification by the neutralisation test and allowed for detailed characterization of the isolates. The predominance of ECHO 30 as the cause of aseptic meningitis is in accordance with the literature data.