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SUMMARYHistorical enquiry into diseases with morbidity or mortality predilections for particular demographic groups can permit clarification of their emergence, endemicity, and epidemicity. During community-wide outbreaks of hepatitis A in the pre-vaccine era, clinical attack rates were higher among juveniles rather than adults. In community-wide hepatitis E outbreaks, past and present, mortality rates have been most pronounced among pregnant women. Examination for these characteristic predilections in reports of jaundice outbreaks in the USA traces the emergence of hepatitis A and also of hepatitis E to the closing three decades of the 19th century. Thereafter, outbreaks of hepatitis A burgeoned, whereas those of hepatitis E abated. There were, in addition, community-wide outbreaks that bore features of neither hepatitis A nor E; they occurred before the 1870s. The American Civil War antedated that period. If hepatitis A had yet to establish endemicity, then it would not underlie the jaundice epidemic that was widespread during the war. Such an assessment may be revised, however, with the discovery of more extant outbreak reports.

(Accepted 7 November 2008; first published online 23 December 2008)SUMMARYSurveillance reports and prevalence studies have indicated that injecting drug users (IDUs)contribute more to the hepatitis C epidemic in the United Kingdom than any other risk group.Information on both the prevalence and incidence of hepatitis C in IDUs is therefore essential tounderstanding the epidemiology of this infection. The prevalence of hepatitis C in specimens fromthe Unlinked Anonymous Prevalence Monitoring Programme collected in 1995, 1996, 1998, 1999,2000, and 2001 was determined using residual syphilis serology specimens from IDUs attending15 genitourinary medicine (GUM) clinics in and outside London. These specimens were tested forantibodies to hepatitis C virus (anti-HCV). Using this cross-sectional design, anti-HCV-negativespecimens were tested for HCV RNA to identify incident infections during the ‘window’ period ofinfection, and thus to estimate HCV incidence. Results of the multivariable analysis showed thatthere was marked variation in prevalence by clinic (P

Inspired by the Ancient Greeks, past students of in fectious diseases have often personified in that many headed monster, Hydra, the terrifying effects of scourges such as cholera, scarlet fever, diphtheria, yellow fever and malaria [1-4]. Hercules-like, these pioneers laboured to prod and probe the beast, in tending to identify its vulnerabilities and deliver the final, mortal blows [5]. They have largely been suc cessful, although cholera and malaria remain as some of the few heads to slay.

The prevalence of concurrent multitypic hepatitis C virus (HCV) infection is uncertain. A sensitive and specific approach to identifying minority HCV genotypes in blood is presented. Following serum extraction and reverse transcription PCR to amplify cDNA originating from the viral 5′ noncoding region, the amplified product mixture was treated with genotype-specific restriction endonuclease to digest the dominant genotype. Residual amplicons were subjected to PCR cloning and then to real-time DNA sequencing using a Pyrosequencer to identify the remaining genotypes. Dilution experiments showed that minority genotypes may be detected when they represent 1:10,000 of the total population and in serum specimens with viral loads as low as 1,000 IU/ml. Of 37 patients with bleeding disorders and 44 injecting drug users, infection by more than one HCV genotype was found in 7 (19%) and 4 (9%) patients, respectively. The low rate of detection in people at high risk of repeated HCV infection suggests that multitypic HCV carriage is uncommon.

Objective: To determine the prevalence and genetic diversity of hepatitis C virus in genitourinary medicine clinic attenders and to assess the extent of sexual transmission of the virus. Methods: A cross sectional, unlinked, anonymous survey in 14 genitourinary medicine clinics situated in England, Wales, and Northern Ireland. Serum specimens from genitourinary medicine clinic attenders, retained as part of the Unlinked Anonymous Prevalence Monitoring Programme (UAPMP) serum archive, were tested in small pools, for the presence of antibody to hepatitis C virus (anti-HCV). The main outcome measures were prevalence of antibodies to hepatitis C virus and identification of hepatitis C virus genotypes. Results: Testing of 17 586 specimens from 1995 showed an adjusted prevalence of anti-HCV in genitourinary medicine clinic attenders of 1.03% (95% CI: 0.89 to 1.16) overall and 0.65% (95% CI: 0.51 to 0.78) among those who did not report injecting drug use. Prevalence in injecting drug users attending genitourinary medicine clinics was 36.9% in both 1995 and 1996. Heterosexual injecting drug users had a higher prevalence of anti-HCV than homosexual/bisexual injectors. The most common hepatitis C genotypes were types 3a and 1a. There was a high degree of concordance between genotype and serotype. Conclusions: The low prevalence of anti-HCV in genitourinary medicine clinic attenders who deny injecting drugs suggests that the majority of hepatitis C infections have been acquired in adult life, mostly by injecting drug use, and that the hepatitis C virus is rarely transmitted sexually. The use of needle exchanges may explain the relatively low prevalence observed in the injecting drug users.

Denaturing gradient gel electrophoresis (DGGE) was used to study the diversity of hepatitis C virus (HCV) quasispecies. Optimized DGGE running conditions were applied to screen for variations in sequences cloned from amplicons originating from the nonstructural 5b (NS5b) gene of HCV in blood of hemophilia patients, intravenous drug users, and blood donors (five specimens from each study group, ca. 40 clones studied per specimen). Clones identified by DGGE as unique were sequenced. NS5b sequence entropy and mean genetic distance in hemophiliacs did not differ significantly from those in the other groups, pointing to a lack of correlation between HCV diversity and the multiplicity of past HCV exposures. DGGE was also applied to investigate variation in the HCV envelope 2/hypervariable region 1 (E2/HVR-1) in serum samples serially taken from two patients during the seroconversion phase of HCV infection. E2/HVR-1 sequence entropy changes were small and not correlated with rising anti-HCV antibody levels, reflecting mutational changes not mediated by antibody selection.

A novel TT virus (TTV)-like DNA sequence was detected in the serum of a patient (PM) with acute non-A–E hepatitis. The full-length genome sequence, referred to here as PM virus (PMV), was obtained and its relationship to other full or near full-length TTV sequences examined. Although it shares a common genomic arrangement and short conserved regions, the majority of the genome is extremely divergent, displaying an average genetic distance of 0·60 from all other TTV sequences. By comparing PMV with TTV genomes representing the most divergent types so far described, six major groups can be distinguished. The level of genetic diversity seen between these genomes is higher than would be expected within a single virus species. Indeed, PMV could be considered the prototype of an independent taxonomic group within the Circoviridae family. A genoprevalence study of sera from blood donors and patients with acute hepatitis suggests that PMV is rare.

A 210-bp DNA segment specific to the human herpesvirus 8 (HHV-8) genome was amplified by nested polymerase chain reaction from 10 of 14 archived oral biopsy samples of HIV-positive patients in London who had no evidence of oral Kaposi's sarcoma (KS). Various oral sites were represented. Oral tissues from 20 general dental patients not known to be HIV-infected were negative. When DNA sequences of these products were compared with sequences derived from 5 oral KS tissues of AIDS patients in London and 10 skin biopsies of Italian patients with Mediterranean KS (total number of positive tissues = 25), 11 were found to be unique. DNA and predicted peptide motifs of these sequences were also different from those in 28 of 36 HHV-8-positive lesions previously reported from American and African patients. HHV-8 is tropic for the oral mucosa of HIV-infected persons, and HHV-8 variants, though diverse, may be geographically restricted.

IgG and IgA antibodies with specificities to a latent and a lytic antigen of human herpes virus 8 (HHV-8) were detectable in the saliva and serum of eight patients whose Kaposi's sarcoma had regressed, seven of whom were HIV-1 infected. The measurement of antibody-specific activity and secretion rate, and the detection of secretory IgA all indicate anti-HHV-8 antibody activity in saliva. The specific humoral responses possibly influence mucosal replication of HHV-8, and in turn, that of HIV.

SUMMARYThe aim of this study was to estimate the amount of childhood hepatitis B virus transmission in children born in the UK, a very low-prevalence country, that is preventable only by universal hepatitis B immunization of infants. Oral fluid specimens were collected from schoolchildren aged 7–11 years in four inner city multi-ethnic areas and tested for the presence of antibody to hepatitis B core antigen (anti-HBc). Those found positive or indeterminate were followed up with testing on serum to confirm their hepatitis B status. The overall prevalence of anti-HBc in children was low [0·26%, 95% confidence interval (CI) 0·14–0·44]. The estimated average annual incidence of hepatitis B was estimated to be 29·26/100 000 children (95% CI 16·00–49·08). The total incidence that is preventable only by a universal infant immunization programme in the UK was estimated to be between 5·00 and 12·49/100 000. The study demonstrates that the extent of horizontal childhood hepatitis B virus transmission is low in children born in the UK and suggests that schools in the UK are an uncommon setting for the transmission of the virus. Targeted hepatitis B testing and immunization of migrants from intermediate- and high-prevalence countries is likely to be a more effective measure to reduce childhood transmission than a universal infant immunization programme.

Hepatitis B virus (HBV) DNA from regions coding for surface and core antigens were amplified and radiolabeled from hepatitis B surface antigen (HBsAg)-positive serum samples by polymerase chain reaction, heat-denatured, and analyzed for conformation-dependent polymorphisms by gel electrophoresis under nondenaturation conditions. Analysis of serum samples representative of diverse and identical HBsAg subtypes showed a wide range of autoradiographic banding patterns, each unique to the specimen. Serial samples of a long-term carrier showed relative stability of banding patterns over time. Epidemiologic analyses using this procedure showed banding patterns of case subjects to be identical to those of persons implicated as the source. This facile and discriminatory approach to the differentiation of viral strains should be useful in the study of HBV transmission.

AIM: To determine if a scheme for validating enzyme immunoassay (EIA) results could be devised that did not require costly and methodically elaborate supplemental assays. METHODS: Samples (n = 525) from patients with haemophilia A, leukaemia, and chronic liver disease and at increased risk of hepatitis C virus infection were tested by EIA-1 (Ortho Diagnostics), an assay which uses recombinant HCV fusion proteins as antigens, and by EIA-2 (United Biomedical), an assay based on synthetic HCV oligopeptide antigens. RESULTS: Samples (n = 193) were repeatedly reactive in both EIAs. Of these, 190 (98%) yielded reactivities in both of two supplemental assays used, one an immunoblot assay (RIBA) using recombinant HCV polypeptides similar to EIA-1 antigens, and the other a neutralisation EIA (EIA-2N) based on antigenic competition with HCV peptides similar to EIA-2 antigens. The three samples not reactive in supplemental tests exhibited low EIA optical density (OD) values (signal/cutoff ratios of less than 3). Hence, all specimens reactive and yielding high OD values in both EIAs were also reactive in supplemental assays. Twenty four samples were reactive in EIA-1 only and nine (38%) of these were reactive in RIBA. Fourteen of the 15 (93%) specimens reactive in EIA-1 but not RIBA were derived from patients with chronic liver dysfunction. Two samples were reactive in EIA-2 only, of which one was reactive in EIA-2N and none in RIBA. CONCLUSIONS: Compared with EIA-2, EIA-1 yielded more validated reactive samples and resulted in more non-validated reactivities. It is therefore suggested that for clinical diagnosis: (i) EIA-1 be used for anti-HCV testing and EIA-2 for validation of EIA-1 reactivities; (ii) samples concordantly reactive in EIA-1 and EIA-2 and displaying high OD readings be considered HCV antibody positive without supplemental testing; (iii) supplemental testing by RIBA be limited to samples reactive in EIA-1 but equivocal or unreactive in EIA-2 and those concordantly reactive but exhibiting low absorbance readings.

Polymorphisms along the hepatitis B virus (HBV) genome have an impact on disease outcome, sensitivity to antiviral treatment, escape from vaccination, and laboratory diagnosis. We have designed a diagnostic tool based on duplex amplification of the whole HBV genome and a high-density DNA chip designed to detect 245 mutations, 20 deletions, and 2 insertions at 151 positions and to determine the genotype of the virus in serum. Assay performances were evaluated with 170 samples, characterized by determination of viral load and sequencing of the Pol, S, and precore genes and the basal core promoter. One hundred fifty-three samples (90%) could be amplified and analyzed by the chip. Only two samples with more than 103genome copies/ml could not be analyzed. Genotype had no impact on analytical sensitivity. Reproducibility studies showed no difference between repeats for codon and genotype determination. Genotype determination by sequencing and the chip were concordant in 148 of 151 samples. Twelve thousand one hundred sixty-one codons were analyzed by both techniques. Only 89.4% could be determined by sequencing, and among the remaining 11,335 codons, 92.8% were identical by sequencing and the chip. Failures to identify an amino acid by the chip were mainly due to reduced hybridization efficiency attributed to unexpected polymorphisms. Optimization of the chip-based reagent for the analysis of the HBV genome is ongoing. This first evaluation showed that DNA chip technology can provide important information in relation to the clinical management of chronic hepatitis B.

Human herpesvirus 8 (HHV-8) open reading frame K1 sequences amplified from the urine of 5 of 78 (6.4%) infected people in Malawi were monotypic. In two people, urinary and oral sequences were genotypically different. Comprehensive evaluation of HHV-8 transmission may require characterization of HHV-8 shed both in urine and orally.

AIM--To evaluate the performance of three newly introduced enzyme immunoassays (EIAs) for hepatitis C virus (HCV) antibodies, based on synthetic oligopeptides as antigens. METHODS--Referred serum samples (n = 173) from people representing groups at high risk of HCV infection were studied. An EIA based on second generation recombinant polypeptide antigens was used for comparison. EIA reactivities were validated by testing repeatedly reactive samples in two recombinant antigen based immunoblot assays. RESULTS--In samples from patients with liver dysfunction and those with bleeding disorders sensitivity of the three peptide based EIAs, manufactured by Innogenetics NV, Biokit SA, and United Biomedical Inc., were all 93%; specificity and efficiency were all greater than 95%. In samples from blood donors (previously tested as positive by the Ortho and Abbott Second Generation EIA) specificity, sensitivity, and efficiency were 95% or greater in all three peptide assays. Sensitivity, specificity, and efficiency of the recombinant antigen based Ortho Second Generation EIA were 100%, 89%, and 93%, respectively, in sera of patients with liver disease and those with bleeding disorders; and 100%, 43%, and 83%, respectively, in prescreened samples from blood donors. CONCLUSION--The peptide EIAs are more specific but less sensitive than the Ortho EIA. Peptide based EIAs should be useful in validating the specificity of Ortho EIA reactivities.

A retrospective case-control study was conducted to determine why some infants born full-term without obstetric intervention to hepatitis B e antigen (HBeAg)-seropositive mothers become infected by hepatitis B virus (HBV) despite having received passive-active immunoprophylaxis. Cases and controls comprised 12 hepatitis B surface antigen (HBsAg)-seropositive infants and 22 HBsAg-seronegative infants, respectively. Infants infected by putative vaccine-escape mutants were excluded. Risk factors, after adjustment for the level of maternal viremia, were the following allelic base changes in maternal HBV:C158, A328, G365, and A479 (P = .017, .005, .003, and .005, respectively). High-level maternal viremia (i.e., > or = 10(8) genome equivalents/mL) was a significant factor only after adjustment for G365 (P = .027). HBV DNA sequences recovered from one of the cases, the case's mother, and three infected contacts all had the high-risk mutations. Specific allelic mutations in maternal HBV and level of maternal viremia are potential predictors of vertical breakthrough infection.

Hepatitis B virus (HBV) infection occurred despite full passive-active immunoprophylaxis in 20 of 321 infants born to mothers seropositive for hepatitis B e antigen. In 2 (12%) of 17 infected infants, mother-infant DNA sequence mismatches were found in a segment of the HBV S gene coding for antigenic determinants of the HBV surface antigen (HBsAg) amplified from sera by polymerase chain reaction (PCR). Point substitutions occurred in codons 120, 134, and 144 of the HBsAg polypeptide in the variant sequence of 1 infant and in codon 126 in the other; all were missense mutations. Mutant sequences could not be recovered from maternal sera by PCR cloning but were selectively generated using an amplification refractory mutation system. The frequency of potential vaccine escape mutants is therefore low, and these preexist maternally as minor variants.

At least five viruses are recognised as causes of acute hepatitis which, in some instances, can develop into chronic disease. The viruses of hepatitis A (HAV) and hepatitis E (HEV) are spread predominantly by the faecal-oral route, whereas hepatitis B, C and D viruses (HBV, HCV and HDV) are spread by blood and other body fluids. The incubation period ranges from about four weeks for HAV to three months for HBV. The diagnosis mainly depends on the detection of various serological markers. The control of infection in the United Kingdom has been facilitated by the introduction of vaccines for HAV and HBV and screening blood for HBV and HCV. Good hygiene and sanitation, the availability of sterile equipment, and measures to modify the behaviour of high-risk groups such as injecting drug users, are also important in the prevention and control of viral hepatitis.

We can confirm the finding of McCrudden et al ( (2000) Gut 46:722–723. [OpenUrl][1][CrossRef][2][PubMed][3][Web of Science][4] ) , that acute infection with hepatitis E virus (HEV) can be acquired in the UK. A 61 year old man presented in March 1999 after a two week illness with fever, malaise, and nausea, followed by cholestatic jaundice and a palpable spleen. Results of tests included alkaline phosphatase 277 IU/l, alanine aminotransferase 2118 IU/l, bilirubin 244 umol/l and INR 1.7. He had no risk factors for liver disease and did not swim in the sea. He had not travelled outside the UK for four years and had never been … [1]: {openurl}?query=rft.jtitle%253DNew%2BEngland%2BJournal%2Bof%2BMedicine%26rft.stitle%253DNEJM%26rft.issn%253D0028-4793%26rft.aulast%253DHutin%26rft.auinit1%253DY.%2BJ.F.%26rft.volume%253D340%26rft.issue%253D8%26rft.spage%253D595%26rft.epage%253D602%26rft.atitle%253DA%2BMultistate%252C%2BFoodborne%2BOutbreak%2Bof%2BHepatitis%2BA%26rft_id%253Dinfo%253Adoi%252F10.1056%252FNEJM199902253400802%26rft_id%253Dinfo%253Apmid%252F10029643%26rft.genre%253Darticle%26rft_val_fmt%253Dinfo%253Aofi%252Ffmt%253Akev%253Amtx%253Ajournal%26ctx_ver%253DZ39.88-2004%26url_ver%253DZ39.88-2004%26url_ctx_fmt%253Dinfo%253Aofi%252Ffmt%253Akev%253Amtx%253Actx [2]: /lookup/external-ref?access_num=10.1056/NEJM199902253400802&link_type=DOI [3]: /lookup/external-ref?access_num=10029643&link_type=MED&atom=%2Fgutjnl%2F47%2F5%2F740.1.atom [4]: /lookup/external-ref?access_num=000078755700002&link_type=ISI

Biotin-labeled Epstein-Barr virus (EBV)-specific DNA probes have been used to detect viral genomes by in situ hybridization. Immunocytochemically amplified signals produced by the hybridized probe allow visualization of viral DNA even in cells previously reported to contain only one or two EBV genomes. In EBV producer lymphoid cell lines, such as B95-8, P3HR-1, or Daudi, activation of latent EBV DNA could be observed in mitotic cells; in non-virus-producing cells of these same lines, EBV was found to be present in low copy numbers. Noninducible cell lines such as IB4, AW-Ramos, and Namalwa exhibited low but clearly positive hybridization. Unexpectedly, significant variations in the amounts of EBV DNA per cell were observed between individual cells of these lines. The EBV DNA in the cloned IB4 cell line was localized to chromosome 4 in metaphase cells, but in the noncloned converted line AW-Ramos, the location of integrated viral DNA was essentially random.

In an eosinophilic population of 47 boys of the same age, a large proportion (92%) were helminth infested or atopic, or both, compared with 36% of 36 controls. The methods used to detect these conditions were not costly or elaborate, except for the radioallergosorbent test, which was used to measure concentrations of circulating IgE antibodies to atopic allergens. It is suggested that an economical approach to detect helminthiasis and atopy in cases of eosinophilia is adopted using the methods employed here, with skin prick tests replacing the radioallergosorbent test.

Salivary gland biopsies from 12 patients with primary Sjögren's syndrome and 10 controls were examined for Epstein-Barr virus (EBV) DNA by in-situ hybridization and for EBV proteins by immunofluorescence and peroxidase techniques. Viral DNA was found in biopsies from two out of 12 patients with primary Sjögren's syndrome and six out of the 10 controls. The DNA and early antigen were in epithelial cells lining the ducts and acini, early antigen expression being limited to the luminal side of the epithelium. In eight biopsies studied with other antibodies, membrane antigen was identified in both acini and ducts but viral capsid antigen and Epstein-Barr nuclear antigen were not detected. EBV was found in biopsies from five of the controls without inflammation or Class II expression. This suggests that, in health, persistence and replication occur without inducing an immune response, possibly due to the restricted expression of early antigen on the luminal of the epithelium, away from immune surveillance. The inflammation in Sjögren's syndrome could be due to a breakdown of this unusual mechanism for viral persistence leading to a vigorous immune response to the virus. However our study provides no evidence to suggest that EBV infection load is increased in this disease.