ABSTRACT
With the changing hepatitis A epidemiology in India, focal viral outbreaks are being reported from different parts of the country. This study presents Hepatitis A Virus (HAV) strain characterization (period 2009-2020) from 18 states of India. For that, blood and stool samples (n â= â280) were screened for HAV RNA and sequences for 5'non-coding and VP3 regions were generated from positive samples (n â= â68). Presence of a single IIIA genotype in all samples indicated IIIA being the only HAV genotype currently circulating in India. Interestingly, it was evident that these strains form two distinct groups suggesting independent evolution of these two clusters.
Subject(s)
Hepatitis A Virus, Human , Hepatitis A , Hepatitis A Virus, Human/classification , Hepatitis A Virus, Human/genetics , Hepatitis A Virus, Human/isolation & purification , India/epidemiology , Genotype , Phylogeny , Feces/chemistry , Feces/virology , Hepatitis A/blood , Hepatitis A/epidemiology , Hepatitis A/virology , Humans , RNA, Viral/analysisABSTRACT
This study determined the prevalence of total hepatitis A antibody (anti-HAV) among 5-7 years old children and their mothers in the whole Cambodia, using a nationwide study, and examined the differences between the two cohorts. A total of 4535 dried blood spot-driven (DBS) samples (2021 mothers and their 2514 children of 5-7 years old) and the concomitant 922 whole blood samples (subset of the whole participants) were collected using a multistage random sampling strategy throughout Cambodia in 2017. Total anti-HAV was detected using the chemiluminescence enzyme immunoassay method. Compared to gold standard whole blood samples, the sensitivity and specificity of DBS mediated anti-HAV detection were 94.8% and 98%, respectively. Total anti-HAV prevalence among mothers was 91.2% (95%CI: 90.0-92.5%), and that of their children was 31.5% (95%CI: 29.7-33.3%). In our study, the low prevalence of total anti-HAV among children indicates the country's improvement of safe water and food supply, hygiene and sanitation. If the hygiene and sanitation are consistently improved in Cambodia, the prevalence might be no longer increased when the children become adults.
Subject(s)
Hepatitis A Antibodies/blood , Hepatitis A Virus, Human/isolation & purification , Hepatitis A/blood , Cambodia/epidemiology , Child , Child, Preschool , Female , Hepatitis A/epidemiology , Hepatitis A/immunology , Hepatitis A Antibodies/immunology , Hepatitis A Virus, Human/immunology , Humans , Male , Prevalence , Seroepidemiologic StudiesABSTRACT
To explore the epidemiology and clinical course of hepatitis A virus (HAV) infections at the Vienna General Hospital. We retrospectively identified patients who were tested positive for HAV-IgM at the Vienna General Hospital form Q1/2008 to Q3/2018. Our definition of severe HAV infection was AST and/or ALT > 5 × above the upper limit of normal (ULN); and liver dysfunction as (i) hepatic encephalopathy or ammonia > 100 µmol/L, (ii) coagulopathy with INR > 1.5, or (iii) jaundice with bilirubin > 5 mg/dL. A total of 578 HAV-IgM (+) were identified, including 31 (5.4%) and 38 (6.6%) without and with liver dysfunction, respectively. A proportional increase in severe HAV cases with and without liver dysfunction occurred in 2016/2017 with (21.5% (vs. 8.0% in the years before; p < 0.001). Thirty-seven (53.6%) patients with severe HAV were hospitalized, 6 (9%) required ICU support, and one patient received liver transplantation within 30 days. Patients with severe HAV and liver dysfunction were more often male (60.5 vs. 43.1%, p = 0.055) and younger (31.5 vs. 63 years, p < 0.001) as compared with other HAV-IgM (+) cases. The observed increase of severe HAV infections in Vienna in 2017 among young males, coincided with a multinational HAV outbreak among MSM. Our data suggests a higher likelihood of severe courses of hepatitis A in MSM. Vaccination against HAV should be recommended for risk groups.
Subject(s)
Disease Outbreaks , Hepatitis A/epidemiology , Adult , Austria/epidemiology , Female , Hepatitis A Virus, Human/isolation & purification , Hospitals, General , Hospitals, Urban , Humans , Male , Middle Aged , Retrospective Studies , Risk Factors , Sexual and Gender MinoritiesABSTRACT
BACKGROUND: Hepatitis A virus (HAV) infection is one of the major causes of acute viral hepatitis. HAV genotypes and its genetic diversity is rarely investigated in our region as well as worldwide. AIMS: The aims of the present study were to determine the HAV genotypes and its risk factors and to investigate the genetic diversity of the HAV isolates in the West Bank, Palestine. STUDY DESIGN: A cohort of 161 clinically and laboratory-confirmed HAV (IgM-positive) cases and 170 apparently healthy controls from all the districts of the West Bank, Palestine during the period of 2014 to 2016 were tested for HAV infection using IgM antibodies, RT-PCR and sequence analysis of the VP3/VP1 junction region of the HAV genome. Phylogenetic analysis, genetic diversity and haplotypes analysis were used to characterize the VP3/VP1 sequences. RESULTS: All the 34 sequences of the HAV were found to be of HAV-IB sub-genotype. The phylogenetic analysis showed four main clusters with cluster III exclusively consisting of 18 Palestinian isolates (18/23-78%), but with weak bootstrap values. A high haplotype diversity (Hd) and low nucleotide diversity (π) were observed. Cluster III showed high number of haplotypes (h = 8), but low haplotype (gene) diversity (Hd = 0.69). A total of 28 active haplotypes with some consisting of more than one sequence were observed using haplotype network analysis. The Palestinian haplotypes are characterized by closely related viral haplotypes with one SNV away from each other which ran parallel to cluster III in the phylogenetic tree. A smaller Palestinian haplotype (4 isolates) was three SNVs away from the major haplotype cluster (n = 10) and closer to others haplotypes from Iran, Spain, and South Africa. Young age, low level of parent's education, infrequent hand washing before meals, and drinking of un-treated water were considered the major HAV risk factors in the present study. CONCLUSION: Haplotype network analysis revealed haplotype variation among the HAV Palestinian sequences despite low genetic variation and nucleotide diversity. In addition, this study reconfirmed that age and parent's level of education as HAV risk factors, while hand washing and treating drinking water as protective factors.
Subject(s)
Hepatitis A Virus, Human/genetics , Hepatitis A/epidemiology , Hepatitis A/virology , Adolescent , Adult , Age Factors , Amino Acid Substitution , Antibodies, Viral/blood , Antibodies, Viral/isolation & purification , Case-Control Studies , Child , Child, Preschool , Cross-Sectional Studies , Educational Status , Female , Genome, Viral/genetics , Haplotypes , Hepatitis A/blood , Hepatitis A/diagnosis , Hepatitis A Virus, Human/isolation & purification , Humans , Immunoglobulin M/blood , Immunoglobulin M/immunology , Male , Middle East/epidemiology , Molecular Epidemiology , Phylogeny , Polymorphism, Single Nucleotide , RNA, Viral/genetics , RNA, Viral/isolation & purification , Risk Factors , Sequence Analysis, DNA , Young AdultSubject(s)
Hepatitis A Virus, Human/physiology , Hepatitis A , Hepatitis E virus/physiology , Hepatitis E , Disease Transmission, Infectious , Global Health , Hepatitis A/epidemiology , Hepatitis A/physiopathology , Hepatitis A/therapy , Hepatitis A/transmission , Hepatitis A Virus, Human/isolation & purification , Hepatitis E/epidemiology , Hepatitis E/physiopathology , Hepatitis E/therapy , Hepatitis E/transmission , Hepatitis E virus/isolation & purification , Humans , Preventive Health Services/methodsSubject(s)
Gastric Dilatation/etiology , Gastroparesis/etiology , Hepatitis A Virus, Human/isolation & purification , Hepatitis A/diagnosis , Jaundice/etiology , Adult , Antibodies, Viral/isolation & purification , Endoscopy, Digestive System , Female , Hepatitis A/complications , Hepatitis A/virology , Hepatitis A Virus, Human/immunology , Humans , Stomach/diagnostic imaging , Tomography, X-Ray ComputedABSTRACT
Acute acalculous cholecystitis (AAC) is the inflammatory disease of the gallbladder in the absence of gallstones. AAC is estimated to represent at least 50% to 70% of all cases of acute cholecystitis during childhood. Although this pathology was originally described in critically ill or post-surgical patients, most pediatric cases have been observed during several infectious diseases. In addition to cases caused by bacterial and parasitic infections, most pediatric reports after 2000 described children developing AAC during viral illnesses (such as Epstein-Barr virus and hepatitis A virus infections). Moreover, some pediatric cases have been associated with several underlying chronic diseases and, in particular, with immune-mediated disorders. Here, we review the epidemiological aspects of pediatric AAC, and we discuss etiology, pathophysiology and clinical management, according to the cases reported in the medical literature.
Subject(s)
Acalculous Cholecystitis/epidemiology , Cholecystitis, Acute/epidemiology , Epstein-Barr Virus Infections/complications , Hepatitis A/complications , Acalculous Cholecystitis/diagnosis , Acalculous Cholecystitis/etiology , Acalculous Cholecystitis/therapy , Anti-Bacterial Agents/therapeutic use , Child , Cholecystectomy , Cholecystitis, Acute/therapy , Cholecystitis, Acute/virology , Epstein-Barr Virus Infections/immunology , Epstein-Barr Virus Infections/virology , Gallbladder/immunology , Gallbladder/surgery , Gallbladder/virology , Hepatitis A/immunology , Hepatitis A/virology , Hepatitis A Virus, Human/isolation & purification , Herpesvirus 4, Human/isolation & purification , Humans , Incidence , Risk Factors , Treatment OutcomeABSTRACT
Since 2015, outbreaks of hepatitis A among men who have sex with men (MSM) have been reported worldwide. To examine the impact of these MSM outbreaks in the Netherlands, we combined notification and epidemiological data with sequence analysis. Our results show the hazards of outbreaks within risk-groups spilling over into the largely susceptible general population. One third of the outbreak-related hepatitis A virus genotypes were detected in non-MSM cases.
Subject(s)
Disease Outbreaks/statistics & numerical data , Hepatitis A Virus, Human/genetics , Hepatitis A/epidemiology , Homosexuality, Male/statistics & numerical data , Adolescent , Adult , Aged , Child , Child, Preschool , DNA, Viral/genetics , Disease Notification/statistics & numerical data , Female , HIV Infections/epidemiology , Hepatitis A/diagnosis , Hepatitis A/transmission , Hepatitis A Virus, Human/classification , Hepatitis A Virus, Human/isolation & purification , Humans , Incidence , Infant , Male , Middle Aged , Netherlands/epidemiology , Sequence Analysis, DNA , Seroepidemiologic Studies , Young AdultABSTRACT
BACKGROUND & AIMS: Acute hepatitis A is transmitted mainly via the faecal-oral route and/or contaminated aliment. Furthermore, several outbreaks in the men who have sex with men (MSM) population classified hepatitis A as a sexually transmitted disease (STD). We aimed to clarify an ongoing hepatitis A outbreak in Barcelona with respect to patients' characteristics and viral phylogenetic analysis. METHODS: We prospectively analyzed 46 cases of hepatitis A infection that were registered in our hospital between January and June 2017. We evaluated demographics data, risk factors, presenting symptoms, sexual orientation, comorbidities and further STD infections. The phylogenetic correlation of the current circulating viruses among them and other hepatitis A strains was assessed by sequencing of the VP1/P2A region. RESULTS: Most patients were male (44, 96%) with median age 33.5 years (range 28-50). Thirty-one (67%) were MSM and 18 (39%) required hospitalization. Molecular phylogenetic analyses revealed that all patients were infected by hepatitis A subgenotype IA strains. Moreover, current strains comprised 3 distinct clusters, previously reported in ongoing outbreaks in the United Kingdom, Berlin and the Netherlands. However, these strains were phylogenetically diverse to those previously reported in Barcelona metropolitan region. CONCLUSIONS: Ongoing hepatitis A outbreak in Barcelona affects primarily the MSM community and is phylogenetically linked to current hepatitis A outbreaks described in other European countries. As a result of the high admission rate, these outbreaks may impact the admission pattern of referral liver units. Control measures, for example vaccinations programs tailored to the MSM community, must be taken to control further spreading.
Subject(s)
Disease Outbreaks/statistics & numerical data , Hepatitis A/epidemiology , Homosexuality, Male , Acute Disease , Adult , Female , Hepatitis A Virus, Human/genetics , Hepatitis A Virus, Human/isolation & purification , Hospitalization/statistics & numerical data , Humans , Male , Middle Aged , Phylogeny , Risk Factors , Spain/epidemiologyABSTRACT
AIM: To investigate the association of chronic hepatitis B and nonalcoholic steatohepatitis with physical fitness in a Taiwanese military male cohort. METHODS: We made a cross-sectional examination of this association using 3669 young adult military males according to cardiorespiratory fitness and hospitalization events recorded in the Taiwan Armed Forces study. Cases of chronic hepatitis B (n = 121) were defined by personal history and positive detection of hepatitis B surface antigen. Cases of nonalcoholic steatohepatitis (n = 129) were defined by alanine transaminase level > 60 U/L, liver ultrasound finding of steatosis, and absence of viral hepatitis A, B or C infection. All other study participants were defined as unaffected (n = 3419). Physical fitness was evaluated by performance in 3000-m run, 2-min sit-ups, and 2-min push-ups exercises, with all the procedures standardized by a computerized scoring system. Multiple linear regression analysis was used to determine the relationship. RESULTS: Chronic hepatitis B negatively correlated with 2-min push-up numbers (ß = -2.49, P = 0.019) after adjusting for age, service specialty, body mass index, systolic and diastolic blood pressures, current cigarette smoking, alcohol intake status, serum hemoglobin, and average weekly exercise times. Nonalcoholic steatohepatitis was borderline positively correlated with 3000-m running time (ß = 11.96, P = 0.084) and negatively correlated with 2-min sit-up numbers (ß = -1.47, P = 0.040). CONCLUSION: Chronic hepatitis B viral infection and nonalcoholic steatohepatitis affects different physical performances in young adult military males, and future study should determine the underlying mechanism.
Subject(s)
Hepatitis B virus/immunology , Hepatitis B, Chronic/physiopathology , Military Personnel/statistics & numerical data , Non-alcoholic Fatty Liver Disease/physiopathology , Physical Fitness/physiology , Adult , Alanine Transaminase/blood , Body Mass Index , Cross-Sectional Studies , Exercise , Hepacivirus/isolation & purification , Hepatitis A Virus, Human/isolation & purification , Hepatitis B Surface Antigens/isolation & purification , Hepatitis B, Chronic/epidemiology , Hepatitis B, Chronic/virology , Humans , Liver/pathology , Male , Non-alcoholic Fatty Liver Disease/blood , Non-alcoholic Fatty Liver Disease/diagnostic imaging , Non-alcoholic Fatty Liver Disease/epidemiology , Prevalence , Retrospective Studies , Taiwan/epidemiology , Ultrasonography , Young AdultABSTRACT
An increased number of hepatitis A cases among refugees, asylum seekers and migrants residing in hosting facilities in Greece were recorded between April and December 2016. In total, 177 laboratory-confirmed symptomatic cases were reported; of these, 149 (84%) occurred in hosting camps mostly among Syrian children under 15 years. All cases reported symptom onset after their entry into the country. Public health interventions focused on hygiene measures and vaccination.
Subject(s)
Disease Notification/statistics & numerical data , Disease Outbreaks/statistics & numerical data , Emigrants and Immigrants/statistics & numerical data , Hepatitis A Virus, Human/isolation & purification , Hepatitis A/epidemiology , Refugees/statistics & numerical data , Adolescent , Adult , Child , Child, Preschool , Communicable Disease Control/statistics & numerical data , Disease Outbreaks/prevention & control , Female , Genotype , Greece/epidemiology , Hepatitis A/diagnosis , Hepatitis A/prevention & control , Hepatitis A Virus, Human/genetics , Humans , Immunization Programs , Infant , Infant, Newborn , Male , Mandatory Reporting , Public Health , Young AdultABSTRACT
We describe a case of acute liver failure (ALF) without hepatic encephalopathy with marked elevation of aminotransferase due to hepatitis A, according to the revised Japanese criteria of ALF. This liver biopsy of the patient showed compatible to acute viral hepatitis and she immediately recovered without intensive care. She had no comorbid disorders. Of interest, phylogenetic tree analysis using almost complete genomes of hepatitis A virus (HAV) demonstrated that the HAV isolate from her belonged to the HAV subgenotype IA strain and was similar to the HAJFF-Kan12 strain (99% nucleotide identity) or FH1 strain (98% nucleotide identity), which is associated with severe or fulminant hepatitis A. Careful interpretation of the association between HAV genome variations and severity of hepatitis A is needed and the mechanism of the severe hepatitis should be explored.
Subject(s)
Aspartate Aminotransferases/blood , Hepatitis A Virus, Human/genetics , Hepatitis A/virology , Liver Failure, Acute/virology , Adult , Biomarkers/blood , Biopsy , Clinical Enzyme Tests , Female , Hepatitis A/diagnostic imaging , Hepatitis A/enzymology , Hepatitis A/pathology , Hepatitis A Virus, Human/classification , Hepatitis A Virus, Human/isolation & purification , Humans , Liver/pathology , Liver Failure, Acute/diagnostic imaging , Liver Failure, Acute/enzymology , Liver Failure, Acute/pathology , Phylogeny , Tomography, X-Ray ComputedABSTRACT
AIM: The aim of this work was to evaluate the efficacy of domestic cooking in inactivating Manila clams experimentally infected with human hepatitis A virus (HAV). METHODS AND RESULTS: Electronic temperature probes were positioned to measure the internal temperature of Manila clams during domestic cooking. Two batches were infected with 10(7) and 10(5) TCID50 ml(-1) of HAV. The infected whole-in-shell clams were divided into three replicates and cooked on a conventional stove both singularly and in group and removed from the pan at fixed intervals. Pools of three digestive glands were examined by virus isolation for three blind passages and cell culture supernatant tested with real-time PCR. CONCLUSION: Results showed that 2-min cooking by a traditional domestic method at a temperature close to 100°C, after the opening up of the valves of all the clams, can completely devitalize the HAV in high viral load-infected clams. SIGNIFICANCE AND IMPACT OF THE STUDY: This is the first study on inactivation of HAV in experimentally infected Manila clams subjected to domestic cooking. At present, labelling all lagoon products as 'requiring cooking before consumption' is highly recommended, but no specifications are given on how long and at what temperature they should be cooked. Considering the high commercial value of Manila clams, our results can provide both the producers and the consumer with useful indications on how to cook clams to prevent the risk of HAV foodborne illness.
Subject(s)
Bivalvia/virology , Cooking/methods , Hepatitis A Virus, Human/growth & development , Shellfish/virology , Animals , Bivalvia/chemistry , Cooking/instrumentation , Hepatitis A Virus, Human/chemistry , Hepatitis A Virus, Human/isolation & purification , Hot Temperature , Humans , Shellfish/analysis , Virus InactivationABSTRACT
BACKGROUND: In 2000, an outbreak of acute hepatitis A was reported in a province adjacent to Bangkok, Thailand. AIMS: To investigate the cause of the 2000 hepatitis A outbreaks in Thailand using molecular epidemiological analysis. METHODS: Serum and stool specimens were collected from patients who were clinically diagnosed with acute viral hepatitis. Water samples from drinking water and deep-drilled wells were also collected. These specimens were subjected to polymerase chain reaction (PCR) amplification and sequencing of the VP1/2A region of the hepatitis A virus (HAV) genome. The entire genome sequence of one of the fecal specimens was determined and phylogenetically analyzed with those of known HAV sequences. RESULTS AND CONCLUSIONS: Eleven of 24 fecal specimens collected from acute viral hepatitis patients were positive as determined by semi- nested reverse transcription PCR targeting the VP1/2A region of HAV. The nucleotide sequence of these samples had an identical genotype IB sequence, suggesting that the same causative agent was present. The complete nucleotide sequence derived from one of the samples indicated that the Thai genotype IB strain should be classified in a unique phylogenetic cluster. The analysis using an adjusted odds ratio showed that the consumption of groundwater was the most likely risk factor associated with the disease.
Subject(s)
Acute Disease/epidemiology , Disease Outbreaks , Feces/virology , Hepatitis A Virus, Human/genetics , Hepatitis A/blood , Hepatitis A/epidemiology , Water Supply , Drinking Water/microbiology , Female , Genome, Viral , Genotype , Hepatitis A/etiology , Hepatitis A/virology , Hepatitis A Virus, Human/isolation & purification , Humans , Male , Odds Ratio , Phylogeny , Polymerase Chain Reaction , RNA, Viral/genetics , Thailand/epidemiologyABSTRACT
BACKGROUND: Hepatitis A virus (HAV) epidemiology in Tunisia has changed from high to intermediate endemicity in the last decades. However, several outbreaks continue to occur. The last reported sequences from Tunisian HAV strains date back to 2006. In order to provide an updated overview of the strains currently circulating in Tunisia, a large-scale molecular analysis of samples from hepatitis A cases was performed, the first in Tunisia. RESULTS: Biological samples were collected from patients with laboratory confirmed hepatitis A: 145 sera samples in Tunis, Monastir, Sousse and Kairouan from 2008 to 2013 and 45 stool samples in Mahdia in 2009. HAV isolates were characterised by nested RT-PCR (VP1/2A region) and sequencing. The sequences finally obtained from 81 samples showed 78 genotype IA and 3 genotype IB isolates. A Tunisian genotype IA sequence dataset, including both the 78 newly obtained IA sequences and 51 sequences retrieved from GenBank, was used for phylogenetic investigation, including analysis of migration pattern among six towns. Virus gene flow from Sfax and Monastir was directed to all other towns; in contrast, the gene flows from Sousse, Tunis, Mahdia and Kairouan were directed to three, two, one and no towns, respectively. CONCLUSIONS: Several different HAV strains co-circulate in Tunisia, but the predominant genotype still continues to be IA (78/81, 96% isolates). A complex gene flow (migration) of HAV genotype IA was observed, with Sfax and Monastir showing gene flows to all other investigated towns. This approach coupled to a wider sampling can prove useful to investigate the factors underlying the spread of HAV in Tunisia and, thus, to implement appropriate preventing measures.
Subject(s)
Genotype , Hepatitis A Virus, Human/classification , Hepatitis A Virus, Human/isolation & purification , Hepatitis A/epidemiology , Hepatitis A/virology , RNA, Viral/genetics , Adolescent , Adult , Aged , Child , Child, Preschool , Cluster Analysis , Feces/virology , Female , Gene Flow , Hepatitis A Virus, Human/genetics , Humans , Male , Middle Aged , Molecular Epidemiology , Molecular Sequence Data , Phylogeny , Polymerase Chain Reaction , Sequence Analysis, DNA , Sequence Homology , Serum/virology , Tunisia/epidemiology , Young AdultABSTRACT
Of the 5 types of viral hepatitis (HAV-HEV), HBV and HCV are by far the most common causes of chronic hepatitis in both pregnant and nonpregnant populations, causing more than 50% of cirrhosis cases and 78% of cases of primary liver cancer. Infection during pregnancy can have adverse effects on both the mother and her fetus. For all 5 viral hepatitis syndromes, early identification allows appropriate measures to be taken to optimize pregnancy outcomes and minimize the risk of perinatal transmission. This article reviews the prevention and management of all 5 viral hepatitis syndromes during pregnancy.
Subject(s)
Hepatitis, Viral, Human/prevention & control , Infectious Disease Transmission, Vertical/prevention & control , Pregnancy Complications, Infectious/prevention & control , Adult , Female , Hepacivirus/isolation & purification , Hepatitis A Virus, Human/isolation & purification , Hepatitis B virus/isolation & purification , Hepatitis Delta Virus/isolation & purification , Hepatitis E virus/isolation & purification , Hepatitis, Viral, Human/diagnosis , Hepatitis, Viral, Human/mortality , Hepatitis, Viral, Human/transmission , Humans , Infant, Newborn , Pregnancy , Pregnancy Complications, Infectious/diagnosis , Pregnancy Complications, Infectious/mortality , Prevalence , Prognosis , Risk Factors , Severity of Illness Index , Viral LoadABSTRACT
BACKGROUND: The hepatitis A virus (HAV) is the most frequent cause of viral hepatitis worldwide and is recognized as one of the most widespread foodborne pathogens. HAV genotypes and subtypes differ in their geographic distribution and the incidence of HAV infection varies considerably among countries, and is particularly high in areas with poor sanitation and hygiene. Phylogenetic analyses are traditionally used in clinical microbiology for tracing the geographic origin of HAV strains. In food microbiology, this approach is complicated by the low contamination levels of food samples. To date, real-time reverse-transcription PCR has been one of the most promising detection methods due to its sensitivity, specificity and ability to deliver quantitative data in food samples, but it does not provide HAV subtyping information. RESULTS: Six subtype-specific RT-qPCR assays were developed for human HAV. The limit of detection of HAV was 50 genome copies/assay for subtype IIB, 500 genome copies assay for IA, IB, IIA and IIIB and 5000 genome copies/assay for IIIA. The specificity of the assays was evaluated by testing reference isolates and in vitro HAV RNA transcripts. No significant cross reactivity was observed. Subtyping results concordant with sequencing analysis were obtained from 34/35 clinical samples. Co-infection with a minor strain of a different subtype was suggested in 5 cases and a recombinant event in one case. CONCLUSIONS: These RT-qPCR assays may be particularly useful for accurately tracing HAV in low-level contaminated samples such as food matrices but also to allow co-infection identification in human samples.
Subject(s)
Genotyping Techniques/methods , Hepatitis A Virus, Human/classification , Hepatitis A Virus, Human/isolation & purification , Real-Time Polymerase Chain Reaction/methods , Reverse Transcriptase Polymerase Chain Reaction/methods , Adolescent , Adult , Aged , Child , Child, Preschool , Female , Food Microbiology , Hepatitis A/virology , Hepatitis A Virus, Human/genetics , Humans , Infant , Male , Middle Aged , Molecular Epidemiology/methods , Sensitivity and Specificity , Young AdultABSTRACT
We tested 19 sera from Austrian patients with acute hepatitis A. A serum from a 48-year-old female patient yielded HAV-nucleic acid that showed 99.7% homology to the HAV-sequence obtained from samples taken during the current outbreak in several European countries, which is associated with consumption of frozen berries. So far, Austria was considered not to be affected by this hepatitis A outbreak.