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1.
Gut ; 2024 Apr 15.
Article in English | MEDLINE | ID: mdl-38621922

ABSTRACT

OBJECTIVES: In high-income countries hepatitis E virus (HEV) is an uncommonly diagnosed porcine-derived zoonoses. After identifying disproportionate chronic HEV infections in persons with cystic fibrosis (pwCF) postlung transplant, we sought to understand its epidemiology and potential drivers. DESIGN: All pwCF post-transplant attending our regional CF centre were screened for HEV. HEV prevalence was compared against non-transplanted pwCF and with all persons screened for suspected HEV infection from 2016 to 2022 in Alberta, Canada. Those with chronic HEV infection underwent genomic sequencing and phylogenetic analysis. Owing to their swine derivation, independently sourced pancreatic enzyme replacement therapy (PERT) capsules were screened for HEV. RESULTS: HEV seropositivity was similar between transplanted and non-transplanted pwCF (6/29 (21%) vs 16/83 (19%); p=0.89). Relative to all other Albertans investigated for HEV as a cause of hepatitis (n=115/1079, 10.7%), pwCF had a twofold higher seropositivity relative risk and this was four times higher than the Canadian average. Only three chronic HEV infection cases were identified in all of Alberta, all in CF lung transplant recipients (n=3/29, 10.3%). Phylogenetics confirmed cases were unrelated porcine-derived HEV genotype 3a. Ninety-one per cent of pwCF were taking PERT (median 8760 capsules/person/year). HEV RNA was detected by RT-qPCR in 44% (47/107) of PERT capsules, and sequences clustered with chronic HEV cases. CONCLUSION: PwCF had disproportionate rates of HEV seropositivity, regardless of transplant status. Chronic HEV infection was evident only in CF transplant recipients. HEV may represent a significant risk for pwCF, particularly post-transplant. Studies to assess HEV incidence and prevalence in pwCF, and potential role of PERT are required.

2.
Epidemiol Infect ; 152: e55, 2024 Mar 15.
Article in English | MEDLINE | ID: mdl-38487841

ABSTRACT

Autochthonous hepatitis E virus (HEV) infection is increasingly reported in industrialized countries and is mostly associated with zoonotic HEV genotype 3 (HEV-3). In this study, we examined the molecular epidemiology of 63 human clinical HEV-3 isolates in Canada between 2014 and 2022. Fifty-five samples were IgM positive, 45 samples were IgG positive and 44 were IgM and IgG positive. The majority of the isolates belong to the subtypes 3a, 3b, and 3j, with high sequence homology to Canadian swine and pork isolates. There were a few isolates that clustered with subtypes 3c, 3e, 3f, 3h, and 3g, and an isolate from chronic infection with a rabbit strain (3ra). Previous studies have demonstrated that the isolates from pork products and swine from Canada belong to subtypes 3a and 3b, therefore, domestic swine HEV is likely responsible for the majority of clinical HEV cases in Canada and further support the hypothesis that swine serve as the main reservoirs for HEV-3 infections. Understanding the associated risk of zoonotic HEV infection requires the establishment of sustainable surveillance strategies at the interface between humans, animals, and the environment within a One-Health framework.


Subject(s)
Hepatitis E virus , Hepatitis E , Swine Diseases , Swine , Animals , Humans , Rabbits , Hepatitis E virus/genetics , Molecular Epidemiology , Canada/epidemiology , Hepatitis E/epidemiology , Hepatitis E/veterinary , Swine Diseases/epidemiology , Genotype , Immunoglobulin G , Immunoglobulin M , Phylogeny , RNA, Viral/genetics
3.
PLoS One ; 18(11): e0291378, 2023.
Article in English | MEDLINE | ID: mdl-37963165

ABSTRACT

BACKGROUND: The rapid spread of HBV has resulted in the emergence of new variants. These viral genotypes and variants, in addition to carcinogenic risk, can be key predictors of therapy response and outcomes. As a result, a better knowledge of these emerging HBV traits will aid in the development of a treatment for HBV infection. However, many Sub-Saharan African nations, including Kenya, have insufficient molecular data on HBV strains circulating locally. This study conducted a population-genetics analysis to evaluate the genetic diversity of HBV among Kenyan blood donors. In addition, within the same cohort, the incidence and features of immune-associated escape mutations and stop-codons in Hepatitis B surface antigen (HBsAg) were determined. METHODS: In September 2015 to October 2016, 194 serum samples were obtained from HBsAg-positive blood donors residing in eleven different Kenyan counties: Kisumu, Machakos, Uasin Gishu, Nairobi, Nakuru, Embu, Garissa, Kisii, Mombasa, Nyeri, and Turkana. For the HBV surface (S) gene, HBV DNA was isolated, amplified, and sequenced. The sequences obtained were utilized to investigate the genetic and haplotype diversity within the S genes. RESULTS: Among the blood donors, 74.74% were male, and the overall mean age was 25.36 years. HBV genotype A1 (88.14%) was the most common, followed by genotype D (10.82%), genotype C (0.52%), and HBV genotype E (0.52%). The phylogenetic analysis revealed twelve major clades, with cluster III comprising solely of 68 blood donor isolates (68/194-35.05%). A high haplotype diversity (Hd = 0.94) and low nucleotide diversity (π = 0.02) were observed. Kisumu county had high number of haplotypes (22), but low haplotype (gene) diversity (Hd = 0.90). Generally, a total of 90 haplotypes with some consisting of more than one sequence were observed. The gene exhibited negative values for Tajima's D (-2.04, p<0.05) and Fu's Fs (-88.84). Several mutations were found in 139 isolates, either within or outside the Major Hydrophilic Area (MHR). There were 29 mutations found, with 37.9% of them situated inside the "a" determinant. The most common mutations in this research were T143M and K122R. Escape mutations linked to diagnostic failure, vaccination and immunoglobulin treatment evasion were also discovered. Also, one stop-codon, W163STP, inside the MHR, was found in one sample from genotype A. CONCLUSION: In Kenya, HBV/A1 is still the most common genotype. Despite limited genetic and nucleotide diversity, haplotype network analysis revealed haplotype variance among HBV genotypes from Kenyan blood donors. The virological properties of immune escape, which may be the source of viral replication endurance, were discovered in the viral strains studied and included immune-escape mutations and stop-codon. The discovery of HBsAg mutations in MHR in all isolates highlighted the need of monitoring MHR mutations in Kenya.


Subject(s)
Hepatitis B virus , Hepatitis B , Male , Humans , Adult , Female , Kenya/epidemiology , Hepatitis B Surface Antigens/genetics , Haplotypes , Blood Donors , Hepatitis B/epidemiology , Hepatitis B/genetics , Hepatitis B/diagnosis , Prevalence , Phylogeny , DNA, Viral/genetics , Mutation , Genotype , Nucleotides , Codon
4.
Emerg Infect Dis ; 29(9): 1890-1894, 2023 09.
Article in English | MEDLINE | ID: mdl-37610234

ABSTRACT

We tested liver samples from 372 Norway rats (Rattus norvegicus) from southern Ontario, Canada, during 2018-2021 to investigate presence of hepatitis E virus infection. Overall, 21 (5.6%) rats tested positive for the virus. Sequence analysis demonstrated all infections to be rat hepatitis E virus (Rocahepevirus ratti genotype C1).


Subject(s)
Hepatitis E virus , Hepatitis E , Animals , Rats , Ontario/epidemiology , Hepatitis E virus/genetics , Hepatitis E/epidemiology , Hepatitis E/veterinary , Genotype
5.
Viruses ; 14(1)2022 01 15.
Article in English | MEDLINE | ID: mdl-35062362

ABSTRACT

Hepatitis A virus (HAV) is an emerging public health concern and there is an urgent need for ways to rapidly identify cases so that outbreaks can be managed effectively. Conventional testing for HAV relies on anti-HAV IgM seropositivity. However, studies estimate that 10-30% of patients may not be diagnosed by serology. Molecular assays that can directly detect viral nucleic acids have the potential to improve diagnosis, which is key to prevent the spread of infections. In this study, we developed a real-time PCR (RT-PCR) assay to detect HAV RNA for the identification of acute HAV infection. Primers were designed to target the conserved 5'-untranslated region (5'-UTR) of HAV, and the assay was optimized on both the Qiagen Rotor-Gene and the BD MAX. We successfully detected HAV from patient serum and stool samples with moderate differences in sensitivity and specificity depending on the platform used. Our results highlight the clinical utility of using a molecular assay to detect HAV from various specimen types that can be implemented in hospitals to assist with diagnostics, treatment and prevention.


Subject(s)
Feces/virology , Hepatitis A virus/genetics , Hepatitis A/diagnosis , Molecular Diagnostic Techniques/methods , Molecular Diagnostic Techniques/standards , Serum/virology , DNA Primers , Disease Outbreaks , Genotype , Hepatitis A/virology , Humans , Limit of Detection , Phylogeny , RNA, Viral , Sensitivity and Specificity
6.
J Infect Dis ; 220(6): 951-955, 2019 08 09.
Article in English | MEDLINE | ID: mdl-30649379

ABSTRACT

Hepatitis E virus (HEV) is a major public health concern in developing countries where the primary transmission is via contaminated water. Zoonotic HEV cases have been increasingly described in Europe, Japan, and the United States, with pigs representing the main animal reservoir of infection. We report an unusual acute hepatitis infection in a previously healthy man caused by a rat HEV with a considerably divergent genomic sequence compared with other rat HEV strains. It is possible that rat HEV is an underrecognized cause of hepatitis infection, and further studies are necessary to elucidate its potential risk and mode of transmission.


Subject(s)
Hepatitis E virus/genetics , Hepatitis E/immunology , Hepatitis E/virology , Immunocompetence , Animals , Genome, Viral , Hepatitis Antibodies/blood , Hepatitis E/transmission , Hepatitis E/veterinary , Hepatitis E virus/classification , Hepatitis E virus/immunology , Hepatitis E virus/isolation & purification , Humans , Immunoglobulin G/blood , Liver/pathology , Male , Middle Aged , Phylogeny , Rats , Zoonoses
7.
J Gen Virol ; 89(Pt 12): 3009-3015, 2008 Dec.
Article in English | MEDLINE | ID: mdl-19008387

ABSTRACT

Hepatitis B virus (HBV) genotype G (HBV/G) is an unusual variant, and little is known about its epidemiology and natural history, particularly the requirement for a co-infecting HBV genotype and their relationship during infection. This study investigated the quasispecies nature of co-infecting genotypes in 39 samples collected over a 6 year period from 13 HBV/G-infected patients. HBV/G infections were found to occur predominantly in males (92 %) and were primarily associated with male homosexual sex (67 %). All patients were infected with HBV/G and HBV/A, or a recombinant HBV/A/G strain. Co-infecting genotypic prevalence was often observed to fluctuate over time, with periods of HBV/G monoinfection in some patients. The average sequence divergence among Canadian HBV/G strains was 1.57+/-0.62 %. Thus, all HBV/G infections in Canada occur in the context of co-infection or recombination with HBV/A, and strains display increased sequence divergence compared with all known HBV/G sequences described to date.


Subject(s)
Hepatitis B virus/classification , Hepatitis B virus/genetics , Hepatitis B/epidemiology , Adult , Canada/epidemiology , Female , Genotype , Hepatitis B/virology , Hepatitis B virus/isolation & purification , Hepatitis B, Chronic/epidemiology , Hepatitis B, Chronic/virology , Humans , Male , Middle Aged , Molecular Sequence Data , Prevalence , Recombination, Genetic , Sequence Analysis, DNA , Viral Load
8.
J Clin Microbiol ; 44(6): 1994-7, 2006 Jun.
Article in English | MEDLINE | ID: mdl-16757589

ABSTRACT

The nucleotide analog adefovir dipivoxil (ADV) is an effective antiviral treatment for chronic hepatitis B virus (HBV) infection, with resistance to ADV estimated to occur less frequently than resistance to lamivudine treatment. The detection of ADV resistance mutations is necessary during therapy to monitor and anticipate possible treatment failure. The INNO-LiPA HBV DR v2 (LiPA; Innogenetics, Ghent, Belgium) is a DNA hybridization line probe assay for the detection of HBV polymerase mutations associated with resistance to lamivudine and ADV. Evaluation of this assay to detect ADV resistance mutations was performed by analyzing 38 patients treated with ADV. Serial samples taken at 6-month intervals during treatment were available for most patients. A total of 124 samples were analyzed by both LiPA and sequencing. By LiPA analysis, 12 patients (31.5%) were found to have mutations associated with resistance to ADV (rtA181V/T and/or rtN236T). This contrasted with sequence analysis, which found nine patients (24%) with either or both mutations. Twice as many samples were rtN236T positive by LiPA (18 of 124) compared to sequence analysis (9 of 124). LiPA detected the rtN236T mutation at least 6 months earlier than its detection by sequencing in patients for whom consecutive serum samples were available. Although less sensitive, sequencing has the advantage of providing information on other polymerase mutations not represented on LiPA strips. The INNO-LiPA HBV DR v2 assay is a very sensitive and specific assay for the detection of the rtN236T mutation associated with resistance to ADV.


Subject(s)
Adenine/analogs & derivatives , Antiviral Agents/pharmacology , Drug Resistance, Viral/genetics , Hepatitis B virus/drug effects , Hepatitis B, Chronic/virology , Organophosphonates/pharmacology , Adenine/pharmacology , Adenine/therapeutic use , Antiviral Agents/therapeutic use , DNA, Viral/analysis , DNA, Viral/isolation & purification , Genotype , Hepatitis B virus/classification , Hepatitis B virus/enzymology , Hepatitis B virus/genetics , Hepatitis B, Chronic/drug therapy , Humans , Microbial Sensitivity Tests , Mutation , Nucleic Acid Hybridization/methods , Oligonucleotide Probes/genetics , Organophosphonates/therapeutic use , RNA-Directed DNA Polymerase/genetics , Reagent Kits, Diagnostic , Reverse Transcriptase Inhibitors/pharmacology , Reverse Transcriptase Inhibitors/therapeutic use , Sensitivity and Specificity , Sequence Analysis, DNA
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