[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.

[Hepatitidis E virus]. / Virus hepatitidy E.

UNLABELLED: The number of detected cases of hepatitis E (HE) shows an increasing trend in the Czech Republic, probably due the introduction of new diagnostic methods and greater awareness of HE. In most cases, the source of infection has remained unconfirmed or entirely unclear. The causative agent, the hepatitis E virus (HEV), belongs to the Hepeviridae family. Recently, many isolates from mammals, birds, and fish have been identified. HEV is widespread among farm pigs and wild boar populations all over the word. Both species serve as HEV reservoirs. Consumption of undercooked meat or offal from the reservoir animals is considered to be unsafe due to zoonotic transmission of HEV. The aim of the present article is to summarize recent findings regarding HEV. KEY WORDS: hepatitis E - zoonosis - food-borne infection.

Comparative hepatitis A seroepidemiology in 10 European countries.

The WHO recommends hepatitis A virus (HAV) immunization according to level of transmission and disease burden. We aimed to identify susceptible age groups by standardized serosurveys to inform HAV vaccination policy in participating countries: Belgium, Czech Republic, England, Finland, Germany, Italy, Lithuania, Malta, Romania, and Slovakia. Each country tested national serum banks (n = 1854-6748), collected during 1996-2004, for anti-HAV antibodies. Local laboratory results were standardized to common units. Forty-one per cent of those aged <30 years and 6% of those aged ≥30 years were susceptible to HAV in Romania; compared to 70-94% and 26-71%, respectively, elsewhere. Romania reported high HAV incidence in children and young adults. Other countries reported HAV disease primarily in older risk groups. The results suggest low level of HAV transmission in most of Europe. Romania, however, appeared as an area with intermediate transmission. Vaccination of risk groups in countries with high susceptibility of young and middle-aged adults needs to be continued.

[Genotypic heterogeneity of hepatitis C virus (HCV) from blood donors in the Czech Republic]. / Genotypová heterogenita viru hepatitidy C (HCV) u dárcu krve v CR.

STUDY OBJECTIVES: Analysis of the genotypic distribution of HCV from blood donors screened in the Czech Republic in 1999-2007 and sequence heterogeneity of HCV isolates in the NS5 region of the HCV genome. MATERIAL AND METHODS: A total of 402 archived serum samples from blood donors collected in 1999-2007 and confirmed positive for anti-HCV antibodies were tested for HCV RNA. In 220 HCV RNA positive sera, PCR was used to amplify a 401-nt fragment of the NS5 region of the HCV genome. The PCR product was sequenced and phylogenetic analysis of the obtained nucleotide sequences was carried out. Genotyping was performed based on the comparison with the sequence data available in the Genbank database. Regression analysis and non-parametric Pearson's chi-square test were used for statistical analysis of the distribution of genotypes by age, sex, sampling place and sampling year. RESULTS: In a representative set of 188 blood donors from the Czech Republic, the following HCV genotypes were determined: 1b (66%), 1a (13.3%), 3 (19.7%), 2a (0.5%) and 2b (0.5%). In 1999-2007, genotype 1b showed a significantly decreasing trend while genotype 3a was becoming significantly more frequent and the incidence of genotype 1a remained unchanged. It was found that the proportion of genotype 1b rises with increasing age of blood donors in contrast to genotypes 1a a 3a. Genotype 1b was detected significantly more frequently in females and genotype 3a significantly predominated in males. Any significant difference was not found in the geographical distribution of HCV genotypes. The mean HCV viral load was 9.6 x 105 I.U./ml. CONCLUSIONS: The predominant HCV genotype among blood donors in the Czech Republic is 1b, showing a significant downward trend in 1999-2007. The increasing prevalence of genotype 3a can be associated with changes in the route of HCV transmission, more precisely with the considerably increased incidence of HCV in injecting drug users over the last 15 years. Surprisingly, the incidence of genotype la remained unchanged over the study period.

[Current situation and trends in the hepatitis C virus genotype distribution among injecting drug users in the Czech Republic]. / Situace a trendy v zastoupení genotypu viru hepatitidy C v populaci injekcních uzivatelu drog.

STUDY OBJECTIVES: To map the current distribution of hepatitis C virus genotypes among injecting drug users (IDUs) in the Czech Republic and to compare the results with the previously reported data. MATERIAL AND METHODS: Of 766 injecting drug users (IDUs) tested in the Remedis hepatology centre, Prague, in 2005 - 2007, 459 (60%) were anti-HCV positive, with viral replication detected by PCR in 326 (71%) of seropositives. PCR products from a randomly selected representative sample of 222 patients were analyzed by sequencing the NS5B region to determine HCV genotypes. Demographic and epidemiological data of the probands were collected by means of an interviewer-assisted questionnare survey. RESULTS: Of 222 study subjects, mostly long-term IDUs, including 154 (69%) males, mean age of 27 years, 131 (61%) reported opiates and 74 (35%) metamphetamine as their favourite drugs. Genotypic analysis found genotype 1, the most common one, in 168 (75.5%) subjects, with slight predominance of subtype 1a, detected in 90 (40.5%) subjects, over subtype 1b, identified in 78 (35%) subjects, while genotype 3 was revealed in 52 (23.5%) IDUs and showed a significant increase in 2007 compared to 2006. The comparison with the control data obtained 5 years earlier showed a substantial rise in the prevalence of genotype 3, previously rare in Czech IDUs, and a significantly increased proportion of subtype 1a among genotype 1 strains. Other genotypes reported from other European countries remain rare in the Czech Republic. CONCLUSIONS: Significant changes were found in the distribution and dynamics of HCV genotypes in the Czech Republic over the last years that are consistent with the changing route of HCV transmission in which injecting drug use currently plays the major role.

The European Sero-Epidemiology Network 2 (ESEN2): standardization of assay results for hepatitis A virus (HAV) to enable comparisons of seroprevalence data across 15 countries.

The European Sero-Epidemiology Network 2 (ESEN2) aimed to compare serological results of vaccine-preventable diseases across Europe. To ensure direct inter-country comparability of hepatitis A virus antibody (anti-HAV) measurements, a standardization panel of 150 sera was developed by a designated reference laboratory and tested by participating national laboratories using assays of choice; each country's results were subsequently regressed against those of the reference laboratory. Quantitatively, the assays were generally highly correlated (R2>0.90). Nevertheless, qualitative comparisons indicated that results obtained with different assays may differ despite the usage of well-established international and local standards. To a great extent standardization successfully alleviated such differences. The generated standardization equations will be used to convert national serological results into common units to enable direct international comparisons of HAV seroprevalence data. The results of this study are expected to contribute to the evaluation and potential improvement of the currently employed immunization strategies for hepatitis in Europe.

The European Sero-Epidemiology Network 2: standardization of assay results for hepatitis B virus.

The aim of the European Sero-Epidemiology Network 2 was to coordinate and standardize the serological surveillance of vaccine-preventable diseases in Europe. In this study, the standardization of hepatitis B virus (HBV) results is described. The 15 participating national laboratories tested a unique panel of 172 sera established by the Greek reference centre for HBV surface antigen (HBsAg), antibodies to HBsAg (anti-HBs) and/or to the HBV core antigen (anti-HBc) by assay methods of their choice. Country-specific quantitative measurements for anti-HBs and anti-HBc were transformed into common units using standardization equations derived by regressing each country's panel results against the reference centre's results, thus adjusting for interassay and interlaboratory variability. For HBsAg, a qualitative analysis (positive/negative) showed at least 99% agreement with the reference laboratory for all countries. By combining these standardized and qualitative results for the markers mentioned earlier, it was possible to achieve comparable estimates of the proportion of the population susceptible to HBV, vaccinated against HBV, with a past HBV infection, and with a current infection or chronic carrier state. Standardization is a very important tool that allows for international serological comparisons to assess the current vaccination policies and the progress of HBV control in Europe.

[Diagnosis and treatment of chronic hepatitis B. Recommendations of the Czech Hepatology Society of the J. E. Purkinje Medical Society and the Society of Infectious Medicine of the J.E. Purkinje Medical Society]. / Diagnostika a lécba chronické hepatitidy B.

Chronic hepatitis B is one of the world's most common infectious diseases. In the Czech Republic it has a prevalence of 0.56%. Antiviral therapy for chronic hepatitis B demonstrably increases quality of life and where indication criteria are met and standard therapeutic procedures are followed, it is clearly cheaper than treatment for the complications of advanced cirrhosis of the liver or hepatocellular carcinoma. At the time of issuing of this recommendation, 4 medicines were classified for the treatment of chronic hepatitis B in the Czech Republic--pegylated interferon (IFN) alpha-2a, conventional IFN alpha, lamivudine (LAM) and adefovir dipivoxil (ADV). In a number of other developed states, entecavir (ETV) and telbivudine (LdT) have also been approved for treatment. The most effective treatment available at present is pegylated IFN alpha-2a, which should be the medication of first choice for initial treatment of hepatitis B, HBeAg positive and negative forms, provided that there are no contraindications for IFN alpha treatment. Conventional (standard, classical) IFN alpha can also be used, though clinical studies have shown it to be less effective than pegylated IFN alpha-2a. The main advantage of interferon compared to other commercially available medications is its relatively shorter and more clearly defined treatment period, the high probability of permanent suppression of virus replication and seroconversion of HBeAg/anti-HBe (in HBeAg positive forms of the illness) and the non-creation of mutant strains of HBV resistant to IFN in the course of treatment. If there are contraindications for IFN alpha (pegylated or conventional) or it is ineffective or poorly tolerated, ADV, ETV, LAM or LdT can be used. LAM and LdT treatments are often accompanied by the appearance of mutant strains of HBV, that are resistant to lamivudine or LdT and therefore they are not preferred.

Long-term protective effect of post-exposure Havrix administration during viral hepatitis Type A outbreaks.

Administration of human normal immunoglobulin (HNIG) post-exposure has been routinely used in Slovakia for outbreak control of hepatitis A, but requires deep intramuscular injection, provides only short-lived protection and is a human blood product. The protective effect of post-exposure administration of an inactivated hepatitis A vaccine was evaluated during 10 outbreaks in Slovakia. Direct contacts of confirmed hepatitis A cases received either: a single dose of hepatitis A vaccine (n = 2171) or immunoglobulin (HNIG, n = 3837). In the HNIG group the number of hepatitis A confirmed cases dropped within the first 7 weeks, however the decrease was not as rapid or as marked as that observed in the vaccinated group where the number of hepatitis A cases dropped within the first 4 weeks after vaccination. Among contacts, 67 cases of hepatitis A were detected during the maximum incubation period of 45 days: 16 cases (0.7%) in the vaccine group and 51 cases (1.3%) in the HNIG group (p < 0.05). After two and three years respectively, 50 and 39 volunteers who had previously received one dose of hepatitis A vaccine received a booster dose and anti-HAV antibodies were measured. Differences in anti-HAV antibody GMCs before and after the booster were statistically significant. The longer time interval (3 years instead of 2) between primary vaccination and booster administration did not seem to impact the magnitude of the booster response. The results of this study show that active post-exposure immunisation with only one dose of inactivated vaccine confers high and long-term protection and effectively controls viral hepatitis A outbreaks.

[Prevalence of hepatitis G virus infection (HGV) in intravenous immunoglobulin recipients in the Czech Republic]. / Výskyt infekce virem hepatitidy G (HGV) u pacientu lécených intravenózními imunoglobuliny v Ceské republice.

The prevalence of hepatitis G virus (HGV) in the serum of intravenous immunoglobulin (IVIG) recipients was studied and risk related to HGV positivity was considered. Although its pathogenicity is unclear, HGV is likely to cause liver disease or lymphoproliferation. Twenty (23%) of 86 tested MG patients were HGV RNA positive. Of the HGV positive patients, three (15%) showed mild elevation of liver enzymes and one (5%) was diagnosed with chronic lymphatic leukaemia prior to the institution of MG replacement. It can be concluded that the HGV prevalence among IVIG recipients is high but is not associated with signs of either liver disease or lymphoproliferation.

[HCV genotyping by combination of the Cobas Amplicor HCV 2.0 test and the reverse hybridization Versant HCV Genotype Assay]. / Vysetrení genotypu HCV pomocí kombinace testu Cobas Amplicor HCV 2.0 a reverzní hybridizace Versant HCV Genotype Assay.

Genotyping of hepatitis C virus (HCV) is of relevance to scheduling the treatment of patients with chronic hepatitis C (VHC), making their prognosis and monitoring the treatment efficacy. A set of 62 sera testing HCV RNA positive in Cobas Amplicor HCV 2.0 test (CA) were genotyped using Versant HCV Genotype Assay (LiPA) Bayer, i.e. the reverse hybridization method, with the CA amplified product being directly used in the assay. Fifty-six out of 57 samples reactive in reverse hybridization (92%) were genotyped. One sample showed a profile differing from any genotype, five samples were not reactive and one sample was not tested within this study design. Two out of five non-reactive sera and one non-tested serum could be genotyped by nested PCR based reverse hybridization. It can be concluded that the CA product resulting from one-step HCV RNA amplification is suitable for use in genotyping by reverse hybridization. The CA product based genotyping procedure is easier to perform, less time-consuming and less costly. The nested PCR based procedure could be used for typing of sera with lower HCV concentrations nontypeable with the combination of CA and Versant HCV Genotype Assay. Forty-eight selected samples were typed not only by reverse hybridization but also by a serological kit Murex HCV Serotyping 1-6 Assay (Abbot Murex). Thirty-seven (77%) of these sera, including all of three sera negative in reverse hybridization, appeared typeable by this kit. Although less sensitive, serotyping may be of relevance to typing of sera with low HCV levels or not containing detectable viral NA which are nontypeable by reverse hybridization. Thirty-three sera appeared genotypeable by both of the methods tested with the results being in good agreement. In two cases only the serotyping method revealed one more type of virus (mixed genotype) compared to the reverse hybridization.

Hepatitis D.

Hepatitis D virus (HDV) is a small, RNA-containing virus that requires the concomitant presence of Hepatitis B virus (HBV) in an obligate manner for its survival and pathogenicity. HDV infection is very uncommon in Czech Republic. The results of antiviral therapy of hepatitis D patients are not satisfactory. Alpha-interferon (alpha-IFN) in high doses (9-10 MU three times a week for 12 months) is usually recommended.

[Hepatitis B virus (HBV) genotypes in the Czech Republic]. / Genotypy viru hepatitidy B (HBV) v Ceské republice.

Hepatitis B virus (HBV) was genotyped based on sequencing and analysis of nucleotide sequences of HBV S gene in sera samples from 176 patients with viral hepatitis B from all over the Czech Republic. The most frequent genotype A was detected in 118 (67.1%) of the patients, followed by genotype D found in 50 (28.4%) of the patients. Genotypes B and C were identified in 6 (3.4%) and 2 (1.1%) of the patients, respectively. The incidence of genotype A statistically significantly increases with age. There is no statistically significant correlation between genotypes A and D and gender or locality (region). Genotypes B and C, found mainly in South East Asia, were first detected in the Czech Republic. Detection of these genotypes in both foreigners and Czech population is suggestive not only of import but also of transmission of these genotypes in the Czech Republic. Fylogenetic analysis documents dissimilarity levels between HBV isolates. Based on the amino sequence derived, the percentages of HBV subtype determinants d (68.6%) and y (31.4%) were established. The gene S sequencing analysis proved suitable for HBV genotyping.

The 2001 serological survey in the Czech Republic--viral hepatitis.

Within serological surveys 2001, prevalence of markers of hepatitis viruses A (anti-HAV), B (anti-HBc, HBsAg, anti-HBs) and for the first time also C (anti-HCV) was investigated. Sera were collected in 2001 and tested by respective kits AxSYM, Abbott. HAV: 2,623 sera were tested for the presence of anti-HAV antibodies. Comparison with serological surveys of 1984 and 1996 revealed again shifts of the age prevalence curve for anti-HAV antibodies towards higher age groups corresponding to time intervals between epidemiological surveys. High prevalence rates of anti-HAV antibodies (more than 20%) were only found for the population age groups who lived in the period of high incidence of VHA, i.e. up to 1965. The prevalence of anti-HAV antibodies increased by about 5-10% in the population under 20 years of age, the increase being significant and assumingly attributable to vaccination against VHA, and remained the same as in 1996 in the age group 20-29 years. HBV: 2,568 sera were tested for the presence of anti-HBc antibodies and 76 reactive specimens were further tested for the presence of HBsAg and anti-HBs antibodies. The prevalence of anti-HBc antibodies continuously increases with age. The total prevalence of anti-HBc antibodies calculated for the Czech population is 5.59% compared to 6.95% recorded in 1996. The calculated prevalence rate of HBsAg is 0.56% and that of anti-HBs antibodies is 3.99% for the non-vaccinee population. HCV: The prevalence rate of anti-HCV antibodies was 0.2% with 6 out of 2,950 sera testing positive. Age dependence could not be assessed because of the small number of positive persons. HCV infection is known to afflict high-risk groups, likely to escape a general serological survey, rather than the normal population.

[Why is there a poor response to combined therapy with interferon alfa 2b and ribavirin in patients with chronic hepatitis C?]. / Proc je odpoved na kombinovanou lécbu interferonem alfa2b a ribavirinem u pacientu s chronickou hepatitidou C tak slabá?

BACKGROUND: Patients with chronic hepatitis C are now treated with a combination of interferon alpha (IFN) and ribavirin. Therapy was used in initial treatment, for those who were resistant to interferon and in relapsed patients. The aim of the study was to evaluate our results and to compare them with the already published data. METHODS AND RESULTS: 27 adult patients of the average age 46 years were cured from 1997 through 1999 for the histologically verified chronic hepatitis C with IFN-alpha 2 and ribavirin. 16 patients suffered from the chronic hepatitis with low, medium or high activity, 11 of them had cirrhosis. Serological testing was done using ELISA-3 test HCV RNA in serum was estimated by the polymerase chain reaction and serotype HCV was identified by the Murex kit. Treatment lasted 6 to 12 months. Incidences of side effects to both remedies, complications in therapy, therapeutical responses and possible failures of the combination therapy were analysed. In 20/27 patients the HCV serotype could be identified and always serotype 1 was found. Average score of histological activity was in patients with serious chronic active hepatitis and cirrhosis 10.1, in other patients 5.6. Sustained biochemical and virological response was found in 5 out of 27 patients (18.5%). Their average age was 39 years. 9 patients (33%) of average age 49 years had no biochemical and virological response. Sustained biochemical response without the accompanying virological response was found at the end of therapy in 10 patients (37%). After the end of therapy, 33% of patients had a relapse. The most frequently seen accompanying sign was the flu-like syndrome, in two female patients we found impairment of thyroid gland function, and in another two the breakthrough phenomenon. Treatment with ribavirin was in seven cases (26%) complicated with rather serious haemolytic anaemia and the dose had to be reduced. Toxic-allergic exanthema in 5 patients (18.5%) represented the second serious complication causing the termination of therapy. CONCLUSIONS: Sustained biochemical and virological response to the combination therapy was found only in patients with mild form of chronic hepatitis C. No such response was found in 8 patients with cirrhosis and in one female patients with medium activity of the chronic hepatitis. The age of patients with sustained response was significantly lower than in patients without the response (39 years versus 49 years). The most frequent serotype was HCV 1. The combination therapy was more frequently accompanied with clinically serious complications. To the adverse effects to the combination therapy belonged in our population the liver cirrhosis, age about 50 or more years, and the genotype HCV 1. It is also the answer to the question why the combination therapy of patients with chronic HCV infection has so poor results.

[GBV-C/HGV, a new virus as one possible cause of hepatitis of unknown etiology]. / Nový virus GBV-C/HGV jako jeden z mozných puvodcu etiologicky nevyjasnených hepatitid.

For etiologically obscure (some 4%) viral hepatitis agents are sought and tested to make elucidation of their cause possible. One of the candidates is since 1995 the newly discovered virus GBV-C/HGV. Despite intense research its relationship to viral hepatitis of obscure origin (VHN) has not been elucidated so far. In the submitted paper the authors attempted to contribute to the elucidation of etiological associations of GBV-C/HGV infection and VHN by comparing the dynamics of markers of the infection in a group of 59 patients with VHN, two control groups exposed to a high risk of parenteral operations and a third comparative group. The first control group comprised 64 patients in a long-term haemodialyzation programme (HD), the second group was formed by 82 patients with haematooncological disease (BD). The third comparative group comprised 22 patients coinfected (CI) with virus of hepatitis C (VHC), or possibly hepatitis B (VHB). The patients with VHN were HBsAg, anti HCV and anti HEV negative. In the majority in the first blood sample transaminases were elevated which was one of the main reasons for examination of GBV-C/HGV RNA. Prevalence of GBV-C/HGV infection, proved by the presence of at least one of the two markers of current or past infection (GBV-C/HGV RNA, antiGBV-C/HGV) was in the compared VHN, HD and BD groups as follows: 88.1%, 59.4% and 43.9%. The frequency of GBV-C/HGV positivity was highest in VHN-76.3%. In control groups HD and BD GBV-C/HGV RNA positivity was substantially lower, 18.8% and 25.6% resp. Long-term continuous viraemia was recorded in patients with VHN in 18.6%. In groups HD and BD it was half that value: 9.3% and 9.18%. In patients with VHN surprisingly after 6.5 months a marked rise of negative findings occurred (5.6x) without the expected increase of antibodies. A similar finding was recorded also in the other groups (HD and BD), incl. CI patients. Disappearance of viraemia was observed most frequently in VHN (55.9%). In groups HD and BD GBV-C/HGV RNA disappeared only in 7.8% and 12.1% resp. In treated patients of the CI group viral RNA was present in 45.5% and it disappeared in 36.4%. On the other hand, seroconversion to antibodies was comparable in VHN, HD and BD (11.9%, 9.4%, 8.5%), only in group CI it was higher (18.2%), obviously in conjunction with treatment of concurrent HCV or HBV infection. Disappearance of viraemia without subsequent seroconversion occurs in GBV-C/HGV infection frequently, the highest rate was observed by the authors in patients with VHN. Disappearance of viraemia does not necessarily imply clearance of GBV-C/HGV but may be due to a change of GBV-C(HGV into a state of persistence without positive laboratory markers of the infection. Persistence of the virus could also be the reason of the assumed conditioned pathogenicity of the virus, and the effect of frequent disappearance of both markers could explain some controversial epidemiological observations when in studies only static data without dynamic associations were used.