Practical use of hepatitis C virus kinetics monitoring in the treatment of chronic hepatitis C

Summary.  Prevention of hepatitis C virus (HCV) infection complications can be achieved by antiviral therapy based on the use of a combination of pegylated interferon (IFN)-α and ribavirin. The steady-state kinetics of HCV infection represents the treatment target. The goal is cure, which is achieved when all infected cells have been cleared from the body. Because of their intrinsic properties, real-time polymerase chain reaction (PCR) methods are rapidly replacing other technologies for routine quantification of HCV-RNA during antiviral therapy. The virological response at week 12 of therapy is currently used to tailor treatment duration in HCV genotype 1 infection only. Recent reports suggest that the virological response at week 4 could be used to tailor treatment duration, whatever the HCV genotype.     

Serotyping of hepatitis C virus in chronic type C hepatitis in Taiwan: Correlation with genotypes

To evaluate the usefulness of a new serologic assay to group hepatitis C virus (HCV), genotypes identified by this serotyping method were compared to those identified by a polymerase chain reaction (PCR) assay with type-specific primers in 71 Taiwanese patients with chronic type C hepatitis. The group-specific antibodies against different HCV genotypes were detected by using an enzyme-linked immunosorbent assay (ELISA) based on group-specific recombinant peptides (C14-1 and C14-2) within the NS4 region. Among 71 patients positive for current second-generation HCV antibodies, HCV RNA was detected in 55 patients by PCR with primers from the 5′ untranslating region, and in 52 by genotype-specific PCR. In 49 (89%) of 55 viremic patients, the results of serotyping by ELISA showed complete agreement with those determined by PCR genotyping, and none of the patients showed a group opposite to that of HCV genotype. The positive rate of group-specific antibodies (69/71;97%) was even better than that of the PCR (55/71;78%). We conclude that this new serotyping assay is highly sensitive and specific for the determination of HCV genotypes, and will be useful in future epidemiologic studies, as well for clinical application.     

Genotypes and multiple infections with hepatitis C virus in patients with haemophilia A in Japan

SUMMARY. Hepatitis C virus (HCV) RNA was tested for, and HCV genotypes determined, in 96 patients with haemophilia A in Japan. Of 88 patients aged ≥ 10 years, 74 (84%) were positive for HCV RNA at a frequency higher than that in patients aged less than 10 years (one of eight, 13% P <0.001). Genotype I/1a was detected in 30 (40%), II/1b in 12 (16%), III/2a in eight (11%), IV/2b in five (7%) and V/3a in 12 (16%); mixed infection with HCV of two different genotypes was identified in the remaining nine (12%). This distribution was markedly different from that in 767 Japanese HCV carriers without haemophilia, in whom II/1b accounted for the majority (68.7%), I/1a was rare (0.5%), V/3a was absent, and mixed infection was observed rarely (1.3%). Mixed infection was transient in all of the seven haemophilic patients who were followed for 1 to 7 years. One of them was infected with genotype II/ 1b and an unclassifiable genotype, which showed nucleotide sequence similarity to genotype 4c from Zaire (82% homology in the El gene) and to 4a from Egypt (91% homology in a part of the NS5b region). In this patient, HCV of genotype II/1b disappeared while that of group 4 survived during a 4-year observation period. These results indicate different epidemiology of HCV genotypes in Japanese haemophiliacs, attributable to HCV contaminating factor VIII imported in the past, and an increased opportunity in haemophiliacs for mixed infection with HCV of different genotypes.     

Hepatitis C virus RNA genome in plasma of patients with non-A,non-B hepatitis

Recently, the assay system of anti-hepatitis C virus antibody (HCV-Ab) was developed. However, there is no clinically useful method to detect hepatitis C virus (HCV) itself. The authors recently developed a method to detect the HCV-RNA genome in plasma using polymerase chain reaction (PCR). In the present study, the specificity of this assay in detecting HCV infection was investigated. Freshly obtained 1 ml plasma specimens from 100 patients with various liver diseases and from 11 control subjects were studied. In patients with non-A, non-B (NANB) hepatitis-related liver diseases, HCV-RNA was detected in 2 out of 7 cases of acute hepatitis, in 29 out of 31 cases of chronic hepatitis, in 17 out of 21 cases of cirrhosis and in 2 out of 6 cases of hepatocellular carcinoma. On the other hand, no HCV-RNA was detected in 15 cases of various types of alcoholic liver diseases, in 12 cases of hepatitis B related liver diseases, and in 11 controls. HCV-RNA was detected in 2 of 6 drinkers with chronic hepatitis. The prevalence of HCV-RNA was not closely related to a history of blood transfusions. These results suggest that our method for HCV-RNA is specific for HCV infection and HCV infection is the likely etiology of most chronic NANB hepatitis cases. The clinical usefulness of our method is illustrated by the fact that we were able to study 100 patients and needed only 1 ml plasma per HCV-RNA assay.     

山西省HBV基因型及亚型分布调查

目的了解山西省乙型肝炎病毒(HBV)的基因型及亚型分布情况。方法对136例乙型肝炎表面抗原(HBsAg)阳性者血清采用PCR-PFLP结合基因型特异性引物-PCR法进行HBV基因型及亚型检测。结果 136例HBV感染者的血清标本经PCR-PFLP分析,B基因型18例,均为Ba亚型,占13.2%;C基因型113例,占83.1%,除4例未分型外均为Ce亚型;D基因型5例,占3.7%。经型特异性引物-PCR分析,18例HBV B基因型中有4例为B/C基因型混合感染;5例经PCR-PFLP确定的D基因型病毒经型特异性引物法分析有2例扩增出E基因型条带。C、B基因型病毒感染者的平均年龄分别为(39.5±13.1)岁和(30.5±14.1)岁,差异有统计学意义(P<0.05);与B基因型相比,C基因型病毒感染者的HBeAg阴转率减慢。结论山西省HBV基因型主要为C(Ce),有少量的B(Ba)和D基因型,且存在B/C混合基因型,且可能存在D和E基因型病毒的重组体;与B基因型比较,感染C基因型病毒更难被机体清除,疾病更易慢性化。     

Transmission of hepatitis C virus by needle-stick injury in community settings

BACKGROUND: Hepatitis C virus (HCV) is predominantly transmitted by blood-to-blood contact, typically by sharing of needles by injecting drug users. Discarded needles could act as a vector for transmission of this infection. METHODS: Two cases of HCV seroconversion following a needle-stick injury in a community setting were identified. The effects of specimen processing and storage conditions on detection of HCV RNA were assessed to provide information about the likelihood of discarded needles containing infectious HCV. RESULTS: Consistent with a role for discarded needles in viral transmission, in vitro studies demonstrated that viral load declined by less than one log following storage for 24 h. CONCLUSION: All needle-stick injuries should be promptly investigated by serology and HCV-PCR.     

High rate of mixed genotypes of hepatitis C virus in patients of an epidemic area in Japan

The subjects of this study were 151 patients (69 males and 82 females) who underwent examination and liver biopsy owing to liver dysfunction in an epidemic area with hepatitis C. Second generation hepatitis C virus antibody (HCV Ab) was positive in 116 (76.8%) of 151 cases. HCV-RNA was detected in 120 (79.5%) by polymerase chain reaction (PCR). In 7 (4.6%) cases, HCV Ab could not be found, but HCV-RNA was detected. Three (2.0%) cases were positive for HCV Ab but negative for HCV-RNA. On the basis of variation in nucleotide sequence within a restricted region in the putative core gene of HCV, HCV genotypes were classed into four types of I, II, III and IV by PCR. The genotypes were identified in 120 cases. Ninety-eight (81.7%) cases carried one of the four types. Type II was found in 76 (63.3%) cases and type III in 22 (18.3%). The other 22 (18.3%) carried simultaneously two different genotypes (mixed type): 21 (17.5%) cases with type II + III and one (0.8%) case with type II + IV. In comparison with the incidence of HCV mixed types in cases with hepatitits C in a non-epidemic area, carriers of mixed types were found at a significantly higher rate in the epidemic area. Liver biopsy of 120 cases with identified HCV genotypes in the epidemic area showed 93 cases of chronic active hepatitis, nine of chronic lobular hepatitis, 10 of chronic persistent hepatitis and eight of liver cirrhosis. No significant correlation could be detected between liver histology and HCV genotypes. Of the 120 cases, 63 (52.5%), 54 (45.0%) and 12 (10.0%) cases had past histories of folk remedies accompanying bleeding, operation and transfusion, respectively. The repetition of these medications may have caused a high ratio of carriers of the mixed genotypes of HCV.     

Hepatitis C virus genotypes in chronic hepatitis C of children

SUMMARY. Several hepatitis C virus (HCV) genotypes have been recently identified and genotype 1b has been correlated with severe liver disease and a poor response to interferon therapy. HCV infection in children is an interesting model for evaluation of the relationship between HCV genotypes and liver disease, because of its relatively short duration and the infrequent association with confounding cofactors. We have investigated HCV genotypes, using a dot-blot hybridization assay with genotype-specific probes, in 36 Italian children with chronic hepatitis C who were otherwise well and had no other underlying disease. Only four patients were symptomatic; liver histology, obtained in 3 3 patients, showed minimal hepatitis in 17 and mild chronic hepatitis in 16. Infection with HCV genotype Ib was found in 55.5% of patients, with a peak prevalence of 83% in children from southern Italy (P < 0.05 vs other regions). The remaining children were infected with HCV genotype la (16.6%), genotype 2 (11.1%). and mixed (10.9%) or undetermined (2.7%) genotypes. In one patient, HCV viraemia was never detected. There was no statistically significant correlation between genotype and age, sex, source of infection, alanine aminotransferase pattern and histological activity index. These results indicate that genotype 1b is widespread among Italian children with chronic hepatitis C, although with significant geographical variations. It is not associated with a more severe liver disease, therefore suggesting that the greater severity of liver disease recently reported in adults could reflect the cumulative effects of disease duration and of interfering cofactors.     

Myasthenia gravis and hepatitis C virus infection

SUMMARY. Chronic hepatitis C virus (HCV) infections are often associated with extrahepatic immunological manifestations, including various autoimmune disorders. The aims of this study were to determine the prevalence of HCV markers in patients with myasthenia gravis (MG) and to determine any relationship with HCV infection. Eighty-three patients with MG, 40 men aged 20–93 years and 43 women aged 13–87 years (mean age 54 years) were studied. The MG patients were positive for antibody to acetylcholine receptor, in addition, their sera was analysed for antibody to HCV (HCVAb) and HCV RNA. HCVAb was detected in two of the 83 patients (2.4%). Four patients were repeatedly HCV RNA positive. They were infected by HCV genotype 1 (one patient), HCV genotype 2a (two patients) and an undetermined HCV genotype in one patient. They received plasmapheresis or intravenous immunoglobulin treatment. Among the four patients, one was infected after the onset of MG without receiving a blood transfusion or using intravenous drugs. The other three had chronic hepatitis C which was discovered at the same time as MG and only one patient had been exposed to blood products. The prevalence of HCV markers in patients with MG (4.8%) was higher than that reported for the general French population, about 1%. This prevalence is similar to that occurring in patients exposed to plasmapheresis or intravenous immunoglobulin. In conclusion, HCV appears to play little, if any, role in causing MG. The higher prevalence of infection among MG patients may be related to transmission in the course of therapy.     

Rare occurrence of occult hepatitis C virus in apparently uninfected injecting drug users: a two‐centre,masked, case–control study

Occult hepatitis C virus (HCV) is a phenomenon where serum HCV RNA is not detected by sensitive commercial assays, but viral RNA is detected by ultrasensitive techniques. Occult HCV infection has not previously been studied in highly exposed, but apparently uninfected (EU) individuals. Two studies examining occult infection in EU subjects were undertaken – an initial two‐centre, masked, case–control study based on cross‐sectional samples (n = 35 subjects) and a single‐centre confirmatory study based on longitudinal samples (n = 32 subjects). Plasma and peripheral blood mononuclear cells were tested for HCV RNA using an ultrasensitive nested polymerase chain reaction assays. Two EU subjects in the first study (10%) and one in the second study (3%) were found to have consistently detectable HCV RNA. Occult HCV infection occurs in high‐risk, apparently uninfected subjects.     

Renal hepatitis C in the absence of detectable serum or hepatic virus

We report an apparently unique case where hepatitis C virus (HCV) RNA was identified in renal tissue from a patient with membranoproliferative glomerulonephritis and treated chronic hepatitis C, despite the absence of detectable virus in the serum or liver (COBAS Amplicor qualitative assay, lower limit of detection 50 IU/ml). The implications of this finding are discussed, with particular reference to current concepts regarding ‘occult’ hepatitis C infection.     

慢性丙型肝炎患者HCV基因型分析

目的研究慢性丙型肝炎患者HCV基因型概况。方法采用基因芯片法检测HCV基因分型;采用PCR法测定HCV RNA定量。结果在570例患者中,HCV RNA阳性552例（95%）,其中1b型400例（72.4%）,2a型63例（11.4%）,3a型20例（3.6%）,3b型20例（3.6%）,1b＋2a型12例（2.1%）,1a型2例（0.4%）,6型7例（1.26%）,1b＋3a型1例（0.18%）,2a＋1b型3例（0.5%）,未定型24例（4.3%）;不同HCV基因型感染者血清HCVRNA水平无统计学差异（P〉0.05）。结论本组患者HCV基因型以1b型为主,2a型次之,多种混合型的出现提示HCV基因型呈现多样化趋势。     

西安地区丙型肝炎病毒基因分型研究

目的　了解西安地区不同人群ＨＣＶ 基因型分布．方法　先用逆转录－ 套式多聚酶链反应检测抗－ ＨＣＶ 阳性血清的ＲＮＡ, 阳性者再用限制性片段长度多态性分析法进行ＨＣＶ 基因分型．结果　１１３ 份ＨＣＶＲＮＡ 阳性血清的基因型如下：输血后丙肝组,Ⅱ型、Ⅲ型及Ⅱ／ Ⅲ混合型分别为８０ ％ ,１６－７ ％ 和３－３ ％ ．散发性丙肝组,Ⅱ型、Ⅲ型分别为７８－９ ％ 和２１－１ ％ ,未检出Ⅱ／Ⅲ混合型． 肝硬变组, Ⅱ型、Ⅲ型及Ⅱ／ Ⅲ混合型分别为８７－９ ％ ,９－１ ％ 和３ ％ ． 原发性肝癌组,Ⅱ型、Ⅲ型分别为８７－１ ％和１２－９ ％ ,未检出Ⅱ／ Ⅲ混合型． 分型总结果为：ＨＣＶⅡ型、Ⅲ型及Ⅱ／ Ⅲ混合型分别为８４－１ ％ ,１４－２ ％ 和１－７ ％ ．结论　ＨＣＶⅡ型是西安地区优势株． 不同人群中ＨＣＶ 基因型总体分布差异无显著性．     

Case report: acute encephalitis immediately prior to acute onset of hepatitis C virus infection

A 34-year-old male presented with acute viral encephalitis of unknown aetiology with subsequent acute onset of hepatitis C virus (HCV) infection. Although the neurological syndrome improved after administration of acyclovir, jaundice appeared. Neurological complications of HCV infections have rarely been described. In this case report, we discuss the close relationship between neurological syndromes and HCV infection. In the future, we hope that further discussion of clinical cases will determine whether or not the HCV produces neurological manifestations.     

Lack of association between occult hepatitis B virus DNA viral load and aminotransferase levels in patients with hepatitis C virus-related chronic liver disease

BACKGROUND AND AIM: Occult hepatitis B virus (HBV) infection in hepatitis C virus (HCV)-infected patients might enhance the severity of chronic liver disease (CLD). To elucidate the correlation between occult HBV infection and the clinical course of HCV-related CLD, we evaluated whether the fluctuation of occult HBV-DNA directly affects the serum alanine aminotransferase (ALT) level. METHODS: Forty-one patients with HCV-related CLD who received regular outpatient treatment and 42 age-, sex-, and antibody to hepatitis B core antigen positivity-matched healthy volunteers were enrolled. Serum HBV-DNA was quantitatively detected using real-time detection polymerase chain reaction (RTD-PCR). Serial serum samples in three patients were measured for HBV-DNA, ALT and HCV core antigen. RESULTS: Hepatitis B virus DNA was amplified in eight of the HCV-related CLD patients (19.5%), which was significantly higher than that of healthy volunteers (2.4%). No significant difference between the genotype 1 HCV-related CLD group and the genotype 2 group was found. Based on the analyses using serial serum samples, the elevation of HBV-DNA did not occur before the ALT flares, but occurred at the same time or after the ALT flares. CONCLUSIONS: The prevalence of occult HBV infection of HCV-related CLD is significantly higher than that of control. Occult HBV infection has no influence on ALT flares among patients with HCV-related CLD.     

Comparison of real-time quantitative polymerase chain reaction with three other assays for quantitation of hepatitis C virus

BACKGROUND AND AIMS: Evaluation of serum levels of hepatitis C virus (HCV) is important for predicting the response to interferon treatment and monitoring its therapeutic efficacy. The aim of this study was to evaluate real-time quantitative polymerase chain reaction (PCR) as a method for the measurement of HCV-RNA. METHODS: The subjects were 50 patients with chronic hepatitis C: 36 with genotype 1b, eight with genotype 2a, and six with genotype 2b. Samples were tested for HCV-RNA by using real-time quantitative PCR with the ABI Prism 7700 sequence detection system, a branched DNA signal amplification assay, and an Amplicor monitor test; and for HCV core protein by using a fluorescent enzyme immunoassay. RESULTS: The detection range of the real-time quantitative PCR was between 10(1)-10(8) copies/mL of HCV-RNA. Hepatitis C virus RNA was detectable in all 50 samples by the use of real-time quantitative PCR, but was undetectable in 14 samples by the use of a branched DNA assay and in two samples by using the Amplicor monitor test; HCV core protein was undetectable in three samples. A significant correlation was found between the results of real-time quantitative PCR and those of the three other assays: branched DNA assay (r = 0.837, P < 0.0001), Amplicor monitor test (r = 0.853, P < 0.0001), and HCV core protein concentrations (r = 0.549, P < 0.0001). CONCLUSIONS: Our results showed that the real-time quantitative PCR was a highly sensitive assay for the measurement of HCV-RNA.     

The genotype analysis of the hepatitis C virus in Heilongjiang Province,China

Introduction: Hepatitis C virus (HCV) infection is a major public health issue. HCV genotype identification is clinically important to tailor the dosage and duration of treatment, and recombination in intra-patient populations of HCV may lead to the generation of escape mutants, as previously observed for other RNA viruses. Up to now, there is no study assessing HCV genotypes and subtypes in Heilongjiang Province, China.Methods: To determine genotype and phylogenetic analysis of HCV in Heilongjiang Province is crucial. In this study, we amplified 3 genome regions (5’UTR, E1, and NS5B) of 30 HCV patients in Heilongjiang Province, amplified products were analyzed by bioinformatics.Results: We found that 23 specimens had concordant subtypes in the 3 gene regions (2a and 1b), 7 HCV patients were considered the recombinants, the recombination pattern of the 7 HCV patients in the 5’UTR, E1, and NS5B region as followed: 1b/2a/1b, 2a/2a/1b, 1b/2a/2a, 1b/2a/1b, 1b/2a/1b, 1b/2a/1b, 2a/2a/1b.Conclusions: The findings in the present study showed that a higher recombination rate (23%) than other researches, and the recombination of 2a/1b in the 5’UTR, E1, and NS5B region was only found in the present study up to now.     

Higher elimination rate of hepatitis C virus among women

SUMMARY. The mortality rate from liver disease in H town of the Fukuoka prefecture in Japan is significantly higher in men than in women. To clarify the gender-related difference, we evaluated subjects with hepatitis B virus (HBV) and hepatitis C virus (HCV) infection in the S area of this town. A total of 824 adults participated in this study, 332 men and 492 women. The incidence of positivity for hepatitis B surface antigen (HBsAg) in serum did not differ significantly between the subjects in the S area (1.9%) and Japanese blood donors (1.5%); however, the incidence of positivity for serum antibody to hepatitis C virus (HCVAb) in the subjects (31.8%) greatly exceeded that in Japanese blood donors (1.3%). The rate of positivity for HCVAb did not differ significantly between men (28.3%) and women (34.1%), but the proportion of serum HCV RNA-positive to HCVAb-positive subjects was significantly higher in men (78.2% in men vs 67.3% in women). The incidence of elevated serum alanine aminotransferase (ALT) in subjects positive for serum HCV RNA was also significantly higher in men (77.0% in men vs 55.8% in women). These results suggest that a more frequent elimination of HCV from serum in women may explain the observed lower mortality from liver disease.     

Prevalence of hepatitis C virus infection in asymptomatic anti-HIV1 negative pregnant women and their children

The prevalence of hepatitis C virus (HCV) infection was studied prospectively in pregnant women in France and their children by detection of anti-HCV with second-generation ELISA (ELISA2). In ELISA2-positive women, anti-HCV was detected with second- and third-generation RIBA (RIBA2 and RIBA3) and serum HCV RNA was detected with PCR. Among 670 women, anti-HIV1-negative, 26 (3.9%) were positive with ELISA2. RIBA2 was positive in 13 and HCV RNA was found in 10. Ten ELISA2-positive women had a further evaluation with assessment of HCV infection in their children. Among the 10 children born to the index pregnancy, only one was positive with ELISA2 and RIBA2 but negative with RIBA3 and PCR; the nine other children were ELISA2, RIBA2, RIBA3, and PCR negative. All 26 siblings (2–16 years old), of whom 14 were born to PCR-positive mothers, were ELISA2 and RIBA2 negative. We conclude that among anti-HIV1-negative pregnant women with normal serum ALT levels, the prevalence of HCV infection is relatively high but the risk for mother-to-infant transmission of HCV seems to be low.