Infection with hepatitis C virus among intravenous-drug users: prevalence,genotypes and risk-factor-associated behaviour patterns in Thailand

Hepatitis C virus (HCV) infection, a major problem worldwide, is usually transmitted parenterally or by use of contaminated needles among intravenous-drug users (IVDU). In a cross-sectional study, demographic data were collected and behaviour patterns investigated in interviews with 453, consenting IVDU. Blood samples were collected from each interviewee and checked for anti-HCV antibodies and, in a PCR-based assay, for the RNA of HCV. Almost all (92.5%) of the IVDU investigated were found positive for anti-HCV and/or the viral RNA. Most (73.5%) of those positive for HCV RNA were found to be infected with genotype 3a alone, the rest being infected with 1b (17.9%), 6a (3.5%), 3b (1.4%), 1a (1.0%), or both 3a and 6a (2.1%) or having non-typable infections (0.6%). Curiously, 26.0% of those who appeared seronegative for anti-HCV were positive for HCV RNA. The longer an interviewee had been using intravenous drugs, the more likely he or she was to be infected with HCV. Among the IVDU, the sharing of needles, syringes and/or other drug-related paraphernalia appeared to be the behaviour carrying the highest risk of HCV infection, giving an adjusted odds ratio and (95% confidence interval) of 4.84 (1.88-12.43). Programmes of needle and syringe exchange should probably be implemented among IVDU in Thailand and elsewhere, to slow the transmission of HCV.     

Impact of the hepatitis B mass vaccination program in the southern part of Thailand.

By 1992, hepatitis B vaccine had been included in the Expanded Program of Immunization (EPI) on a nation-wide scale in Thailand. With the results now available from Songkhla Province in the south of Thailand, we are able to fully evaluate its impact on the prevalence of HBV infection and carrier rate. The population studied comprised 180 randomly selected children aged between 2 months and 15 years who had attended Hat Yai hospital due to any acute illness affecting neither the liver nor the immune system. Their sera were examined for the hepatitis markers HBsAg, anti-HBs and anti-HBc, respectively, using a commercially available test kit. We detected anti-HBs in 106 of the 180 children (58.9%) with its prevalence peaking within the age groups of 0-2 (94.4%) and 3-5 years (75.6%), respectively. Six children, five within the age groups of 6-10 and 11-15 years showed anti-HBc, one of them was diagnosed as a chronic carrier; the sixth one of the age group of 0-2 years most probably displayed passive maternal antibodies. The overall HBV carrier rate amounted to 0.55%. The hepatitis B mass vaccination program has proved highly efficient in protecting newborns from infection and heralds the promise of eventually eradicating hepatitis B virus in the not so far future.     

Immunogenicity and safety of a new inactivated hepatitis A vaccine in Thai young adults

In view of the increasing median age of hepatitis A virus (HAV) infection observed recently in Asia, and the resulting increased number of symptomatic cases occurring in adults, with the concomitant risk of outbreaks, immunization against this agent on a national scale might be considered. An open clinical trial was conducted in Thai adolescents and young adults in order to establish the immunogenicity and safety of a new inactivated hepatitis A vaccine. At 24-week intervals, two doses (primary dose and booster) of the hepatitis A vaccine (160 antigenic units per dose) were administered to 80 HAV-seronegative healthy volunteers, their ages ranging from 16 to 25 years. Local and systemic reactions were recorded within the first 7 days after each injection. Anti-hepatitis A virus antibody concentrations were measured by a modified radioimmunoassay before and one month after each injection. No serious adverse reactions were reported. Local reactions were confined to transient pain at the injection site, occurring within 24 hours after injection in 42.5% of the subjects after the first dose and 24.1% of the patients after the booster dose. Systemic reactions (particularly asthenia or myalgia) were observed in 35.0% and 8.9% of subjects after the first and the booster injection, respectively. Most of these reactions were transient. One month after the first dose, all 78 formerly seronegative subjects had attained satisfactory seroconversion levels of anti-HAV antibody concentrations (> or = 20 mIU/ml) which they maintained until the booster. The booster dose elicited a 21-fold increase of HAV antibody levels, with a geometric mean titer of 2,964 mIU/ml (95% CI, 2,467-3,560), indicative of long-term protection. This new inactivated hepatitis A vaccine appears to be safe and highly immunogenic upon administration of a primary dose followed by a booster dose after 24 weeks. In countries where socio-economic improvement has postponed hepatitis A infection from early childhood (mostly asymptomatic) towards adolescence and adulthood, with the symptoms increasing in severity, inclusion of inactivated hepatitis A vaccine in a preventive vaccination program might be of benefit.     

Hepatitis B virus genotypes and hepatocellular carcinoma in Thailand

AIM: The role of hepatitis B virus (HBV) genotypes on the clinical features and prognosis of patients with hepatocellular carcinoma (HCC) is currently unknown. The aim of the present study was to evaluate the distribution of HBV genotypes and their clinical relevance in Thai patients. METHODS: HBV genotypes were determined by PCR-RFLP in stored sera of 93 asymptomatic carriers, 103 patients with chronic hepatitis, 60 patients with cirrhosis and 76 patients with HCC. The clinical data were analyzed in relation to the HBV genotype. RESULTS: HBV genotypes C and B were predominant in Thailand, accounting for 73% and 21%, respectively. The distributions of genotypes B and C were similar in HCC patients compared to the other groups. Genotype C was significantly more common in HCC patients who were under 40 years old than genotype B (18% vs 0%, P= 0.03), but was significantly less common in patients older than 60 years (26% vs 56.5%, P= 0.01). The positive rate of hepatitis B e antigen (HBeAg) in patients with genotype C was significantly higher than that in patients with genotype B (71.6% vs 44.4%, P= 0.03 in chronic hepatitis; 56.8% vs 11.1%, P= 0.01 in cirrhosis). There were no differences between HCC patients with genotypes B and C regarding tumor staging by CLIP criteria and the overall median survival. Multivariate analyses showed that HBV genotype was not an independent prognostic factor of survival in HCC patients. CONCLUSION: Patients with genotype C had a higher positive rate of HBeAg and exhibited earlier progression of cirrhosis and HCC than those with genotype B. However, there were no differences in the risk of developing HCC and its prognosis between patients with these genotypes.     

Molecular characterization of hepatitis-A-virus infections,in the context of two outbreaks in southern Thailand

As hepatitis A virus (HAV) is usually transmitted through the faecal-oral route, hepatitis A is a communicable disease. In countries of intermediate to low endemicity, sudden outbreaks of human infection with the virus may occur. Between September 2001 and April 2002, there were two outbreaks of HAV infection in the Ruso and Yeengor districts of Narathiwas province, in southern Thailand. Isolates of HAV were recovered during these outbreaks, from 14 in-patients with acute hepatitis in Ruso (12 positive for anti-HAV IgM and all positive for HAV RNA), 16 children with asymptomatic infection in Yeengor (14 positive for anti-HAV IgM and nine for HAV RNA), and four isolated cases in Bangkok (all positive for anti-HAV IgM). Molecular characterization of the VP1-P2A region of each isolate was followed by phylogenetic analysis. All of the isolates from Narathiwas province were found to be of genotype 1a, to have the same VP1 nucleotide sequence, and to show a high level of sequence homology (>/= 99.5%) with the isolates from Bangkok and with previous Thai isolates. These results should facilitate further research into HAV transmission and genotype identification in community outbreaks.     

Molecular epidemiological study of hepatitis B virus in Thailand based on the analysis of pre-S and S genes.

Aims: This study was undertaken to determine the prevalence and characteristics of hepatitis B virus (HBV) genotypes, antigen subtypes, "a" determinant variants and pre-S gene mutations circulating on a large scale in Thailand. Methods: The sequences of the Pre-S1, Pre-S2 and S regions were determined in serum samples of 147 HBsAg and HBV DNA-positive subjects who had been enrolled from the nationwide seroepidemiological survey conducted on 6213 individuals in 2004. Results: The results showed that genotypes C, B and A accounted for 87.1%, 11.6% and 1.3%, respectively. The distribution of the HBV antigen subtypes was: adr (84.4%), adw (14.2%) and ayw (1.4%). Regarding the "a" determinant, 2/43 (4.65%) and 2/104 (1.92%) samples of vaccinated and non-vaccinated subjects, respectively, displayed mutations, all ofwhich were Thr126Asn. Sequencing analysis showed the pre-S mutations in 14 (9.5%) samples, with pre-S2 deletion as the most common mutant (4.1%) followed by pre-S2 start codon mutation (2.9%), both pre-S2 deletion and start codon mutation (2.0%), and pre-S1 deletion (0.7%). The pre-S mutations were associated with older age and higher mean serum HBsAg level. Conclusion: This study demonstrated that HBV genotype/subtype C/adr and B/adw were the predominant strains circulating in Thailand. The "a" determinant variants seemed to be uncommon, and might not be attributed to vaccine-induced mutation.     

Hepatitis viruses and chronic liver disease

We have investigated several groups of Thai patients diagnosed with chronic liver disease including chronic hepatitis, cirrhosis and hepatocellular carcinoma, as well as cholangiocarcinoma, for the prevalence of infection with either one of the hepatitis viruses B, C, G and the novel hepatitis virus TT (TTV). The 168 patients tested comprised 120 men and 48 women with their median age ranging from 42.3 to 62.3 years. Screening for antibodies to HBV and HCV was performed by a commercially available serological test kit, for the presence of HBV and TTV DNA by PCR, and of HCV and HGV RNA by RT-PCR, respectively. There was a clear two-fold higher prevalence of HBV (49%) over HCV (27%) infection and a four-fold higher frequency compared to HGV (13%) and TTV (11%) infection, respectively, in those individuals with chronic hepatitis, cirrhosis, and hepatocellular carcinoma, whereas all but one patient with cholangiocarcinoma the etiology of which has been ascribed to parasitic infestation, were free of all viral markers. In Thailand chronic HBV, and to a lesser extent, chronic HCV infection represent the two most common causes of hepatitis potentially proceeding to chronic liver disease, whereas the clinical significance pertinent to HGV and TTV remains to be elucidated.     

Prevalence of antibodies to parvovirus B 19 in Thailand

Infection with human parvovirus B 19, a single-stranded non-enveloped DNA virus of worldwide distribution, is rather common and displays a broad spectrum of clinical manifestations of varying severity, depending on the patient's immune response. As the target of infection are the erythroid precursor cells, patients can experience an aplastic crisis. Usually, at least in immunocompetent individuals, viremia ceases with the appearance of virus-specific antibodies in the patient's serum whereupon the patients retain lifelong immunity to reinfection. Since data as to the prevalence of this agent has not been established for Thailand, the purpose of the present study was to investigate its frequency among 3 distinct groups, comprising 30 healthy children. 64 children with acute unrelated illness, and 35 voluntary blood donors, respectively, by means of enzyme linked immunosorbent assay. Our results have shown that, as reported for other countries, anti-parvovirus IgG increases in an age-dependent manner and is established at an overall prevalence of 20.16%, inviting the conclusion that the local population is infected by this agent as frequently as those of other countries in the Far East. Further studies need to be undertaken in order to elucidate its prevalence among members of high-risk groups.     

Humoral immune response following hepatitis B vaccine booster dose in children with and without prior immunization.

One hundred and twenty-three children who had received no, incomplete and complete primary hepatitis B vaccination but had negative or very low anti-HBs titer were immunized with a single dose of recombinant hepatitis B vaccine. Blood tests for anti-HBs were obtained at 30 +/- 5 days after the booster immunization. Twelve of 18 (66.7%) children without prior immunization (group 1) seroconverted following the single dose Seroconversion rates in children who had undetectable anti-HBs with incomplete and complete primary immunization (group 2 and 3) were 83.34% and 94.5%, respectively. All children with complete 3- dose vaccination but who had low anti-HBs titer (group 4) also seroconverted. This study confirmed that immunological memory, allowing a protective anamnestic response, lasted at least 8 years in children who had received primary HB immunization with undetectable anti-HBs. Therefore, we conclude that the booster dose after complete vaccination is not necessary in healthy children.     

Transfusion transmissible virus TTV and its putative role in the etiology of liver disease

TTV, the transfusion transmissible hepatitis virus infects mainly patients at risk for parenteral exposure and hence, prone to develop chronic liver disease, as well as healthy populations worldwide. Most TTV infections appear to occur parenterally, with viremia detected frequently in blood donors and blood products. The substantial proportion of asymptomatic individuals never exposed to blood-borne agents, and its high prevalence among healthy subjects implicates the fecal-oral route as another potential for transmission. According to the TTV DNA levels detected in liver tissue, it apparently replicates in hepatocytes, and TTV DNA is present in sera of patients with posttransfusion hepatitis of unknown etiology closely correlated with ALT levels. However, TTV initiating the development of chronic liver disease or causing posttransfusion hepatitis could not be confirmed, as most patients positive for TTV DNA remain asymptomatic and those progressing towards chronic liver disease are invariably coinfected with either the hepatitis B or C virus. Also, TTV coinfection does not aggravate the symptoms associated with hepatitis B or C. Similarly, it does not cause posthepatitis aplastic anemia, and high-risk patients can immunologically clear the viral DNA. In conclusion, being widely distributed and apparently nonpathogenic, TTV might represent an opportunistic but innocent virus reminiscent of hepatitis G virus, with a negligible role in the etiology of chronic liver disease.