HCV genotype shift occurred over the 15 years in PWIDs in the Czech Republic.

OBJECTIVES: To follow on the epidemiology of HCV, especially genotypes spreading among people who inject drugs (PWID) in Prague and surrounding Central Bohemia, Czech Republic. METHODS: 546 patients who reported past and/or recent injecting of drugs were recruited in the years 2010-2012. They were initially tested for anti-HCV. Real-time PCR was used for quantification and genotyping of hepatitis C virus. Obtained data from the years 2010-2012 were compared with historical controls from periods of 1998-2000 and 2005-2007. RESULTS: Of 546 initially recruited and tested patients were 393 (72%) anti-HCV seropositive and of them 269 (68.4%) had detectable HCV PCR RNA. The most prevalent subtype was 3a in 97 patients (36.1%), 1a was detected in 85 patients (31.6%) and 1b in 57 patients (21.2%). These three genotypes were responsible for nearly 89% of infections. CONCLUSION: Significant increase in both genotypes 1a and 3a over the 15 years was apparent and significant, followed by the decrease in genotype 1b. In the genotype 1b and genotype 3a the significance has risen with the years of data collection. Described genotypic shift reflects the evolution of HCV epidemics and corresponds with the mode of transmission.

A high prevalence of viral hepatitis C in a socially excluded Roma community in Brno.

OBJECTIVES: To conduct a pilot study to assess the prevalence of blood-borne and sexually transmitted diseases (STDs) in a social-ly excluded Roma community engaging in risky behaviours in Brno, Czech Republic. METHODS: Fifty subjects engaging in risky behaviours, of whom 35 self-reported to belong to the Roma ethnicity, were recruited while receiving treatment in a newly established addiction centre in Brno between March and December 2017. All subjects were tested for blood-borne diseases and STDs. Epidemiological and demographic data were collected by means of assisted interview at the time of the first contact. RESULTS: Forty-three (86%) of 50 participants were anti-HCV positive. Of 35 subjects from the Roma subgroup, 32 (91.5%) tested anti-HCV positive. Among the 43 anti-HCV positive subjects, 35 (81.4%) also showed HCV RNA positivity. Of the 32 anti-HCV positive in the Roma subgroup, 26 (81.3%) were HCV RNA PCR positive. Only HCV (hepatitis C virus) genotypes 1 (a,b) and 3 were detected in the study group. Nine Roma subjects (25.9%)were newly diagnosed with syphilis of which none of them was aware. All study patients were negative for anti-HIV 1,2. CONCLUSION: As a proof of concept, this pilot study showed the importance of targeting epidemiological research and preventive care at excluded communities engaging in risky behaviours. The high anti-HCV seroprevalence in the Roma population in Brno who self-admitted intravenous drug use as well as the nine newly diagnosed cases of syphilis illustrate not only a high prevalence of risky behaviours in this excluded community but also the absence of systematic health care coverage in this population.  A positive point is that when an appropriate model of care is used, even the Roma clients are willing and able to comply with the therapy. This is true of both viral hepatitis C and syphilis: thanks to close cooperation with addictology services and opiate substitution treatment, all nine patients successfully completed 2 weeks of anti-treponemal antibiotic treatment. More systematic work with socially excluded communities including specific models of care tailored to the needs of poorly compliant patients is an essential prerequisite for controlling HCV epidemics in the Czech Republic. An additional effect in the surveillance of other infectious diseases linked to risky behaviours can be considered as an added value.

[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.

Structure and functions of hepatitis C virus proteins: 15 years after.

Since its discovery in 1988, the hepatitis C virus (HCV) has become a hot topic of research by many groups around the world. This globally spread infectious agent is responsible for a large proportion of chronic viral hepatitides. The clue to halting the hepatitis C pandemic may be the detailed understanding of the virus structure, its replication mechanism, and the exact functions of the various proteins. Such understanding could enable the development of new antivirals targeted against hepatitis C virus and possibly an effective vaccine. This review recaps the current knowledge about the HCV genome 15 years after its discovery. The structure and function of particular viral structural (core, E1, E2) and nonstructural (NS2, NS3, NS4, NS5) proteins and noncoding regions known to date are described. With respect to frequent conflicting reports from different research groups, results reproducibly demonstrated by independent investigators are emphasized. Owing to many obstacles and limitations inherent in doing research on this noteworthy virus, the current knowledge is incomplete and the answers to many important questions are to be expected in the future.

[Distribution of HBV genotypes in the Czech population--pilot study]. / Distribuce genotypu HBV v ceské populaci--pilotní studie.

A compact structure of the hepatitis B virus genome does not provide much space for development of mutations capable of further replication. In spite of that, 7 genotypes have been described so far. Discovery of the hepatitis B virus (HBV) genome diversity led to a range of further questions. Individual genotypes are subjects of investigation these days as for the pattern of disease transmission, course of disease, and consequences of an infection. The aim of this pilot study was to describe genotype distribution in the infected population of the Czech Republic. Moreover, epidemiological and clinical characteristics were monitored in our sample. We studied a group of 65 patients with positive HBsAg from which 48 samples of patients were genotyped. PCR (polymerase chain reaction) method was used in order to amplify S region of the HBV genome. Direct sequencing of PCR products was used, this is the most accurate method for genotype identification. A relatively uniform genotypic distribution was identified within the study population--only 2 genotypes, A and D, were found. Genotype A was identified in 35 (73%) patients and genotype D in 13 (27%) patients. No statistically significant association between the particular genotype and epidemiological and clinical parameters was proved in this pilot study. This study brings basic information on genotype distribution. A possible proof of correlation with clinical and epidemiological characteristics will require examination of a larger sample of patients.

Genotypic distribution of TT virus (TTV) in a Czech population: evidence for sexual transmission of the virus.

BACKGROUND: TTV is a new DNA virus distinguished by its high degree of strain heterogeneity. The geographic clustering of viral genotypes suggests frequent community transmission. While no specific human disease has yet been linked to it, a transmission mechanism that facilitates strain diversity may eventually select for a strain that will become pathogenic. OBJECTIVE: This study was performed to examine the prevalence, genotypic distribution, and mode of transmission of TTV in detail. STUDY DESIGN: Three groups of study subjects were recruited between October 1998 and January 2000 in Prague, Czech Republic. Group 1 included 152 injection drug users with liver disease; group 2 included 102 persons with liver disease who denied ever using injection drugs; group 3 included 111 prospective blood donors. TTV DNA was detected from blood by a semi-nested PCR assay, and a selected set of PCR products was genotyped by direct sequencing. Factors associated with TTV prevalence in groups 1 and 2 subjects were compared. RESULTS: TTV was detected in 15.8, 13.7, and 13.5% of Groups 1, 2, and 3 subjects, respectively (P>0.05). The most common genotype was 2 (54%), followed by 1 (13%). The prevalence of TTV viremia was nearly three times higher in persons with a present or past history of hepatitis B compared to those without (P<0.05). TTV prevalence increased proportionately with the number of lifetime sex partners in both groups (P<0.05); it was highest (32%) among non-users of injection drugs who had five or more lifetime sex partners. CONCLUSION: TTV prevalence in the Czech population is similar among blood donors, persons with liver disease, as well as in a high-risk population of injection drug users. TTV appears to be sexually transmitted.

Genotypic and epidemiologic characteristics of hepatitis C virus infections among recent injection drug user and nonuser populations.

The recent surge in injection drug use (IDU) in Prague, Czech Republic, provided an opportunity to compare the distribution of hepatitis C virus (HCV) genotypes among recent injection drug users and nonusers. We provide evidence that the most common HCV subtype, 1b, has been circulating in Prague longer than the other subtypes but that IDU practice has magnified its recent prevalence, and that the primary drug of choice among IDU subjects appears to influence the prevalence of a particular HCV subtype.

Nested restriction site-specific PCR to detect and type hepatitis C virus (HCV): a rapid method to distinguish HCV subtype 1b from other genotypes.

Genotypic differentiation of hepatitis C virus (HCV) has become an integral part of clinical management and epidemiologic studies of hepatitis C infections. Thus, it is extremely important in areas such as the Czech Republic, where current instrumentation and kits for assessing HCV infection are too costly for widespread use. We describe a new and relatively inexpensive method called nested restriction site-specific PCR (RSS-PCR) that generates a "fingerprint" pattern to represent an HCV genotype without the use of restriction endonucleases and that specifically differentiates HCV genotype 1b from the other HCV genotypes. The RSS-PCR method was applied directly to serum samples from patients with hepatitis C from the Czech Republic and from patients with known HCV genotypes from the United States. The method was validated by comparison of the subtype determined by RSS-PCR to the subtype determined from analysis of the 5' noncoding region (NC) or the nonstructural protein gene (NS5b) nucleotide sequence of HCV in these clinical samples. From 75 Czech samples containing HCV RNA, three distinct RSS-PCR patterns were observed; 54 were predicted to contain subtype 1b, 19 were predicted to contain subtype 1a, and 2 were predicted to contain subtype 3a. Among 54 samples predicted to contain HCV genotype 1b, all were confirmed by their 5' NC or NS5b sequences to be subtype 1b. Thus, both the sensitivity and specificity of the RSS-PCR test for the differentiation of HCV subtype 1b from the others were 100%. While the assay described here was designed to specifically differentiate HCV subtype 1b from the other HCV genotypes, the RSS-PCR method can be modified to differentiate any HCV genotype or subtype of interest. Its simplicity and speed may provide new opportunities to study the epidemiology of HCV infections and the relationship between HCV genotypes and clinical outcome by more laboratories throughout the world.