Genotype IIIA Research Articles

Genotypes of hepatitis a virus in Turkey: first report and clinical profile of children infected with sub-genotypes IA and IIIA

BackgroundHepatitis A virus (HAV) is a food and water-borne virus causing clinical (mainly hepatitis) and subclinical disease in humans. It is important to characterize circulating strains of HAV in order to prevent HAV infections using efficacious vaccines. The aim of this study was the detection and characterization of the circulating strains of HAV in Turkey by performing serology, RT-PCR, sequencing and phylogenetic analysis.MethodsIn this study, 355 HAV suspected cases were analysed by ELISA for the presence of antibodies to HAV. RNA was extracted from 54 HAV IgM positive human sera. None of the suspect cases were vaccinated against HAV and they never received blood transfusions. Samples found positive by RT-PCR using primers targeting the VP1/VP2A junction and VP1/VP3 capsid region of HAV, were subjected to sequencing and phylogenetic analyses.ResultsIgM type antibodies to HAV were detected in 54 patients. Twenty one of them were students. The age of IgM positive cases was between 3 and 60 years. IgM positivity differed in age groups and was higher in the age group 3 to 10 years. Phylogenetic analysis showed that the majority of HAV strains detected in this study belong to the “HAV 1B” cluster. In addition, the HAV sub-genotypes IA (KT874461.1) and IIIA (KT222963.1) were found in 2 children. These sub-genotypes were not previously reported in Turkey. The child who carried sub-genotype IIIA travelled to Afghanistan and presented with abdominal pain, icterus and vomitus. He was positive for anti-HAV IgM and IgG but negative for hepatitis B and C. Liver enzymes like aspartate aminotransferase, alanine aminotransferase, alkaline phosphatase, gamma-glutamyl transferase and lactate dehydrogenase were severely elevated. Bilirubin levels were also increased. White blood cells, neutrophils and hemoglobin were decreased while lymphocytes and monocytes were increased. Similar clinical signs and laboratory findings were reported for the child infected with sub-genotype IA but aspartate aminotransferase and alanine aminotransferase were not severely elevated.ConclusionsThe results indicate that molecular studies determining the HAV genotype variation in Turkey are timely and warranted. The majority of IgM positive cases in 3–10 year old patients indicate that childhood vaccination is important. Sub-genotype IB is the most prevalant genotype in Turkey. Surprisingly, sub-genotype IA and IIIA are also present in Turkey; future diagnostic efforts need to include diagnostic methods which can identify this emerging HAV genotypes. Our results also show that one important risk factor for contracting hepatitis A virus is international travel since genotype IIIA was detected in a child who had travelled to Afghanistan.

Prototheca zopfii isolated from bovine mastitis induced oxidative stress and apoptosis in bovine mammary epithelial cells.

Bovine protothecal mastitis results in considerable economic losses worldwide. However, Prototheca zopfii induced morphological alterations and oxidative stress in bovine mammary epithelial cells (bMECs) is not comprehensively studied yet. Therefore, the aim of this current study was to investigate the P. zopfii induced pathomorphological changes, oxidative stress and apoptosis in bMECs. Oxidative stress was assessed by evaluating catalase (CAT), superoxide dismutase (SOD), glutathione peroxidase (GPx), malondialdehyde (MDA) contents and lactate dehydrogenase (LDH) activity, while ROS generation and apoptosis was measured by confocal laser scanning microscopy. The results revealed that infection of P. zopfii genotype II (GTII) significantly changed bMECs morphology, increased apoptotic rate and MDA contents at 12 h (p < 0.05) and 24 h (p < 0.01) in comparison with control group, in time-dependent manner. LDH activity and ROS generation was also increased (p < 0.01) at 12 h and 24 h. However, SOD and CAT contents in bMECs infected with GTII were decreased (p < 0.05) at 12 h, while GPx (p < 0.01), SOD (p < 0.05) and CAT (p < 0.01) levels were reduced at 24 h. In case of GTI, only CAT and GPx activities were significantly decreased when the duration prolonged to 24 h but lesser than GTII. This suggested that GTII has more devastating pathogenic effects in bMECs, and the findings of this study concluded that GTII induced apoptosis and oxidative stress in bMECs via the imbalance of oxidant and antioxidant defenses as well as the production of intracellular ROS.

The sirtuin inhibitor sirtinol inhibits hepatitis A virus (HAV) replication by inhibiting HAV internal ribosomal entry site activity

Epigenetics plays a role in the regulation of gene expression. Epigenetic changes control gene expression at the transcriptional level. Our previous study suggested that the La protein, which is mainly localized in the nucleus, was associated with hepatitis A virus (HAV) internal ribosomal entry site (IRES)-mediated translation and HAV replication. The aim of this study was to investigate whether epigenetic compounds have effects on HAV IRES-mediated translation and HAV replication. Sirtinol, a sirtuin inhibitor, inhibited HAV IRES-mediated translation in COS7-HAV-IRES cells. Treatment with 10 μM sirtinol resulted in a significant reduction in the intracellular RNA levels of HAV HA11-1299 genotype IIIA in Huh7 cells. Epigenetic treatment with a sirtuin inhibitor may represent a new treatment option for HAV infection. In conclusion, epigenetic control was involved in HAV IRES-dependent translation and HAV replication. Special attention should also be paid to underlying viral diseases in the clinical use of epigenetic treatments for malignancies.

Molecular epidemiology of hepatitis A virus infection in Northeast India.

The present study was undertaken to screen the molecular epidemiology of Hepatitis A virus (HAV) in Northeast India (NEI) who are ethnically distinct, tribal dominated and of lower socio-economic status with almost no information available from NEI on these aspects. Briefly, 3 ml blood was collected from 324 random liver disease cases with jaundice, receiving care at Central Hospital, N.F. Railway, Guwahati, Assam with informed consent. The patients detected with HAV-IgM positive status were included and were stratified as acute viral hepatitis (AVH) and fulminant hepatitis (FHF) based on clinical profile. Viral RNA was isolated and HAV-RNA was detected by Real-time PCR using primers for the VP3-VP1 region. HAV genotyping was studied by PCR-direct sequencing-phylogenetic analysis approach using the VP1/2A region of HAV isolates. Statistical analysis was performed using SPSS13.0 software. A total of 69 cases were HAV infected with two HBV co-infected cases (n = 69 + 2 = 71), 62 cases and two co-infected cases were AVH and others were FHF cases. HAV infection was predominant in especially in the young and adult age group. HAV-RNA was detected in 28 cases, out of which 19 cases could be genotyped (12 AVH, 7 FHF); which showed the prevalence of genotype IIIA or IA only. Although HAV genotype IIIA was the major genotype in both the AVH (10/12, 83.33%) and FHF (5/7, 71.43%) group, but the difference in distribution of genotypes in AVH and FHF cases was statistically non-significant (P = 0.550). HAV genotype IIIA is associated with the majority of HAV infected cases and severity in NEI.

Hepatitis A virus genotype distribution during a decade of universal vaccination of preadolescents.

A universal vaccination program among preadolescents was implemented in Catalonia, Spain, during the period of 1999–2013 and its effectiveness has been clearly demonstrated by an overall significant attack rate reduction. However, reductions were not constant over time, and increases were again observed in 2002–2009 due to the occurrence of huge outbreaks. In the following years, in the absence of large outbreaks, the attack rate decreased again to very low levels. However, an increase of symptomatic cases in the <5 age group has recently been observed. This is an unexpected observation since children younger than 6 are mostly asymptomatic. Such a long vaccination campaign offers the opportunity to analyze not only the effectiveness of vaccination, but also the influence of the circulating genotypes on the incidence of hepatitis A among the different age groups. This study has revealed the emergence of genotype IC during a foodborne outbreak, the short-lived circulation of vaccine-escape variants isolated during an outbreak among the men-having-sex-with-men group, and the association of genotype IIIA with the increase of symptomatic cases among the very young. From a public health perspective, two conclusions may be drawn: vaccination is better at an early age, and the vaccination schedule must be complete and include all recommended vaccine doses.

The JAK2 inhibitor AZD1480 inhibits hepatitis A virus replication in Huh7 cells

The JAK2 inhibitor AZD1480 has been reported to inhibit La protein expression. We previously demonstrated that the inhibition of La expression could inhibit hepatitis A virus (HAV) internal ribosomal entry-site (IRES)-mediated translation and HAV replication in vitro. In this study, we analyzed the effects of AZD1480 on HAV IRES-mediated translation and replication. HAV IRES-mediated translation in COS7-HAV-IRES cells was inhibited by 0.1–1 μM AZD1480, a dosage that did not affect cell viability. Results showed a significant reduction in intracellular HAV HA11-1299 genotype IIIA RNA levels in Huh7 cells treated with AZD1480. Furthermore, AZD1480 inhibited the expression of phosphorylated-(Tyr-705)-signal transducer and activator of transcription 3 (STAT3) and La in Huh7 cells. Therefore, we propose that AZD1480 can inhibit HAV IRES activity and HAV replication through the inhibition of the La protein.

Outbreak of hepatitis A in an orphanage in Pune city, India

Aims: An outbreak of hepatitis was reported in an orphanage, located in the Pune city, India. Investigations were carried out to identify the etiological agent, source of infection and subclinical infections Study design: Descriptive study. Place and Duration of Study: An orphanage, located in the Pune city, India. Study was carried out in August - September 2014. Methodology: The children in the orphanage were investigated for hepatitis. Clinical, epidemiological findings were recorded and sanitary survey was done. Sera collected from children were tested for anti HAV IgM antibodies, anti HEV IgM antibodies, HBsAg, and serum alanine amino transferase levels. Stools were processed for HAV RNA detection, sequencing and phylogenetic analysis. Results: Fourteen suspected jaundice cases were line listed (boys:7, girls:7). Overall clinical attack rate was 15.1% (14/93). Among all 93 resident children, blood samples 83.9% (78/93) and stool samples 67.7% (78/93) could be investigated. Presence of anti HAV IgM antibodies were detected in 20 children (14 symptomatic and 6 asymptomatic). Presence of HAV RNA detected in 16 stool samples and all HAV RNA sequences clustered with genotype IIIA. Potential source of the infection was probably food served to children. Conclusion: An outbreak of HAV in children was confirmed by serology and virus detection by PCR in stool samples. It would be advisable to vaccinate all children above one year of age in an orphanage set ups.

Co-infection with hepatitis C virus (HCV) in Estonian intravenous drug users HIV epidemic

Estonian concentrated HIV epidemic started in August 2000 when rare HIV-1 CRF06_cpx was introduced into the population of intravenous drug users (IDUs). The majority of these HIV-positive subjects are likely co-infected with HCV but the prevalence of HCV and its genotypes during HIV concentrated epidemic in Estonia is largely unknown. The Estonian HIV database collects clinical and laboratory data (including HCV) of HIV-positive patients on medical care. Of 4500 patients on medical care 3500 are entered into Estonian HIV database (in total 8664 diagnosed). Aim: to describe the prevalence of HCV infection and the distribution of the HCV genotypes among HIV-positive subjects infected during HIV concentrated epidemic in Estonia. Data for present analyses was extracted from Estonian HIV database on 2nd of January 2014 and it comprised subjects diagnosed HIV-positive from 2000. In total 2,420 of 3,476 (70%) HIV-positive subjects were HCV antibody positive and 1,184 (64%) were HCV RNA positive. More than half of HCV-positives were men (66%) and the median age was 32 years (inter quartile range 30-36 y). The prevalence of HCV was higher in subjects verified to be HIV-positive between 2002 and 2010 as compared to between 2011 and 2013 (90% – 60% in 2002 – 2010 vs 50% – 45% in 2011 – 2013; p <0.05). The prevalence of HCV-positivity was equally high in subjects reporting the use of intravenous drugs (88%), the use of other narcotics but intravenous drugs (69%) and persons who have not reported the use of illegal drugs (71%). In total 640 subjects had HCV genotype data available. The dominating genotypes were Ib (53%) and IIIa (36%), however, in recent years the prevalence of genotype Ia is raising (from 2.6% in 2005 to 25% in 2013). The distribution of HCV genotypes between different IDUs and non-IDUs was similar. Altogether 5% of HCV RNA positive subjects (66% with genotype Ib or IIIa; 29% with mixed or unknown; 5% with Ia or II) received HCV treatment and all except one admitted the drug usage. Of 59 subjects 48 (81%) received both HCV and HIV therapy. Decreased prevalence of HCV-positive subjects among HIV-positives may suggest a lowered HIV transmission through intravenous route during the last years. The high prevalence of HCV in persons who did not report the use of illegal drugs might indicate under-reported drug usage in this population.

Genotypic shift of the hepatitis A virus and its clinical impact on acute hepatitis A in Korea: a nationwide multicenter study

The genotypic shift of hepatitis A virus (HAV) and its correlation with clinical course has not been evaluated in acute hepatitis A (AHA). From June 2007 to May 2009, we prospectively enrolled 546 AHA patients. We performed a nested reverse transcriptase polymerase chain reaction (RT-PCR) using the serum samples in addition to phylogenetic analysis, then we compared patient clinical features. Among 351 successfully genotyped patients, we found genotype IIIA in 178 patients (51%) and IA in 173 patients (49%). The sequences of genotype IA are identical to previously reported Korean genotype IA, and the new IIIA genotype is closely related to NOR24/Norway. We retrospectively analyzed 41 AHA samples collected from 2000 to 2006 and found that all of them were genotype IA. Patients with genotype IIIA showed significantly higher levels of aspartate aminotransferase, higher levels of alanine aminotransferase, and lower platelet counts than patients with genotype IA when comparing baseline laboratory data or peak/lowest laboratory data during the disease course. However, there were no differences in duration of hospital stay, incidence of cholestatic hepatitis, acute kidney injury, and acute liver failure, or mortality between them. A genotypic shift of the HAV was identified in Korean AHA subjects, and genotype IIIA HAV has become endemic. Although there were significant differences in the biochemical responses of AHA between genotype IA and genotype IIIA patients, we did not detect any differences in clinical outcomes such as complications or mortality.

Increased circulation of hepatitis A virus genotype IIIA over the last decade in St Petersburg, Russia

The current study, covering the period 2004-2009, is a part of long-term monitoring for hepatitis A virus (HAV) strains circulating in St Petersburg, Russia. The HAV RNA was isolated directly from the sera of hepatitis A patients and RT-PCR was carried out using primer pairs for VP1/2A and VP1 genomic regions. PCR products were sequenced and 324 nucleotides from VP1/2A and 332 from the VP1 region were used for phylogenetic analysis. The results show that the IA subtype was the most common circulating subtype during the follow-up period, as found in the previous study: almost 90% of the isolated HAV strains belonged to the IA subtype. The large hepatitis A food-borne outbreak in St Petersburg in 2005 was caused by HAV IA. However, the proportion of HAV isolates belonging to subtype IIIA significantly increased in the period 2001-2009 (7.9%) compared to the period 1997-2000 (none found). The subtype IIIA was first found in St Petersburg in 2001 among a group of intravenous drug users. The increase in its circulation during the decade suggests that this previously unusual genotype has been permanently introduced into the general population of St Petersburg. These results indicate the usefulness of molecular epidemiological methods for studying changes in the circulation of HAV strains.

Genetic Analysis of Hepatitis A Virus Strains That Induced Epidemics in Korea during 2007–2009

Hepatitis A virus is one of the most prominent causes of fecally transmitted acute hepatitis worldwide. In order to characterize the viral agents causing an outbreak in Korea (comprising North and South Korea) from June 2007 to May 2009, we collected specimens and performed genotyping of the VP1/P2A and VP3/VP1 regions of hepatitis A virus. We then used a multiple-alignment algorithm to compare the nucleotide sequences of the 2 regions with those of reference strains. Hepatitis A virus antibodies were detected in 64 patients from 5 reported outbreaks (North Korea, June 2007 [n = 11]; Jeonnam, April 2008 [n = 15]; Daegu, May 2008 [n = 13]; Seoul, May 2009 [n = 22]; and Incheon, May 2009 [n = 3]). We found 100% homology between strains isolated from the Kaesong Industrial Region and Jeonnam. While those strains were classified as genotype IA strains, strains from Seoul and Incheon were identified as genotype IIIA strains and showed 98.9 to 100% homology. Genotype IIIA was also dominant in Daegu, where strains were 95.7 to 100% homologous. All hepatitis A virus strains isolated from the Kaesong Industrial Region, Jeonnam, Seoul, and Incheon belonged to a single cluster. However, strains from Daegu could be classified into 2 clusters, suggesting that the outbreak had multiple sources. This study indicates that hepatitis A virus strains of 2 different genotypes are currently cocirculating in Korea. Moreover, it documents an increasing prevalence of genotype IIIA strains in the country.

Outbreak of hepatitis A in an extended family after importation by non-immune travellers

The incidence of hepatitis A in England has declined in recent years, but travel-related cases and imported infections remain a challenge. We report an outbreak of hepatitis A in an extended family where two primary cases were infected while in Pakistan and two secondary cases were infected in England. All four were infected by the same genotype IIIA virus. Testing of the children in the extended family by dried blood spots (DBS) determined that three had evidence of recent past infections (anti-HAV IgM positive), one had a current asymptomatic infection (anti-HAV IgM and HAV RNA positive) and one was incubating the virus (anti-HAV IgM negative, HAV RNA positive). HAV RNA from the DBS was identical to the adult cases. This outbreak demonstrates secondary spread of hepatitis A by asymptomatic children after importation from abroad and highlights the importance of preventing travel-associated hepatitis A infection.

Hepatitis A Virus genotype and its correlation with the clinical outcome of acute hepatitis A in Korea: 2006–2008

Korea has recently experienced a nationwide outbreak of hepatitis A. This study aimed to investigate hepatitis A virus (HAV) genotypes and to compare clinical features between patients infected with HAV genotype IA and those with genotype IIIA. From September 2006 to August 2008, 595 patients with symptomatic hepatitis A were enrolled prospectively in four hospitals in Korea. Among them, 556 patients participated in this study by providing serum or stool samples for genotypic analysis. HAV RNA was detected in 499 patients (89.7%). Major genotypes included IA (n = 244, 48.9%) and IIIA (n = 244, 48.9%), and the remaining genotype was IB (n = 11, 2.2%). From September 2006 to August 2007, the distribution of genotypes IA and IIIA were 64.6% and 35.6%, respectively, which changed to 42.3% and 54.6%, respectively, from September 2007 to August 2008, indicating change of circulating HAV genotypes in the study period from IA to IIIA. Major patterns of amino acid substitution in the VP3/VP1 junction region were observed at position 512 (P → L) in genotype IA and at 520 (R → K) in genotype IIIA. Patients with genotype IIIA infection showed significantly higher aminotransferase levels, prothrombin time, and leukocyte count, with more severe symptoms than those with genotype IA at the time of admission. These results suggest the occurrence of a change of circulating HAV genotypes in recent community-wide outbreaks of hepatitis A in Korea, and genotype IIIA infection, compared with genotype IA infection, might show more severe clinical manifestations.

Comparative analysis of disease severity between genotypes IA and IIIA of hepatitis A virus

Although hepatitis A is a major health problem worldwide, it has not yet been clarified whether or not viral factors affect the clinical characteristics. This study aimed to investigate if a genotype of hepatitis A virus (HAV) affects disease severity among adolescent and adult populations. Clinical data and specimens were collected from patients ≥16-years-of-age with acute hepatitis A at two university hospitals in Korea during the two study periods: 1998 and 1999 (n = 45), and 2009 (n = 66). Nucleotide sequencing of the complete VP1 region of the HAV isolates was performed for phylogenetic analysis and genotyping. Clinical parameters related to disease severity were compared by HAV genotype to determine its clinical relevance. Of the 87 patients, 47 were male and the mean age was 29.8 ± 8.1 years. The genotype IIIA (93.0%, 53/57) was predominant in the year 2009, whereas IA (93.3%, 28/30) was the major genotype in 1998 and 1999. When comparing disease severity between the two HAV genotypes, the patients with genotype IIIA were older and had higher alanine aminotransferase (ALT) levels, prolonged prothrombin times and lower serum albumin levels. In a multivariate logistic regression model, higher ALT levels ≥ 1,000 IU/L (odds ratio [OR] 11.7, 95% confidence interval [CI] 2.5-54.0) and longer hospitalization (OR 22.49, 95%CI 4.6-132.5) were associated independently with genotype IIIA. In conclusion, this study indicates that HAV genotype might be one of the viral factors responsible for the disease severity of hepatitis A.

Immunological and molecular epidemiological characteristics of acute and fulminant viral hepatitis A.

BackgroundHepatitis A virus is an infection of liver; it is hyperendemic in vast areas of the world including India. In most cases it causes an acute self limited illness but rarely fulminant. There is growing concern about change in pattern from asymptomatic childhood infection to an increased incidence of symptomatic disease in the adult population.ObjectiveIn-depth analysis of immunological, viral quantification and genotype of acute and fulminant hepatitis A virus.MethodsSerum samples obtained from 1009 cases of suspected acute viral hepatitis was employed for different biochemical and serological examination. RNA was extracted from blood serum, reverse transcribed into cDNA and amplified using nested PCR for viral quantification, sequencing and genotyping. Immunological cell count from freshly collected whole blood was carried out by fluorescence activated cell sorter.ResultsFulminant hepatitis A was mostly detected with other hepatic viruses. CD8+ T cells count increases in fulminant hepatitis to a significantly high level (P = 0.005) compared to normal healthy control. The immunological helper/suppressor (CD4+/CD8+) ratio of fulminant hepatitis was significantly lower compared to acute cases. The serologically positive patients were confirmed by RT-PCR and total of 72 (69.2%) were quantified and sequenced. The average quantitative viral load of fulminant cases was significantly higher (P < 0.05). There was similar genotypic distribution in both acute and fulminant category, with predominance of genotype IIIA (70%) compared to IA (30%).ConclusionsImmunological factors in combination with viral load defines the severity of the fulminant hepatitis A. Phylogenetic analysis of acute and fulminant hepatitis A confirmed genotypes IIIA as predominant against IA with no preference of disease severity.

Molecular and Clinical Characterization of Hepatitis A Virus in Gwangju and Jeonnam Province

Dominant genotype of hepatitis A virus (HAV) in Korea had been known to be genotype 1A in 1990s. Recently, the epidemiologic change of HAV genotype was reported with an upsurge of acute hepatitis A in Korea. The aim of this study was to investigate the change of HAV genotypes and clinical characteristics in Gwangju and Jeonnam province. From November 2008 to October 2009, a total of 82 patients with acute hepatitis A were enrolled prospectively. HAV genotype was determined using reverse transcriptase polymerase chain reaction and sequencing of the PCR products of VP1/2A region of HAV. HAV RNA was detected in 82% (67/82). The genotype IIIA (88%, 59 cases) was significantly more frequent than genotype IA (12%, 8 cases) (p ≤ 0.01). The subtypes of genotype IIIA were AJ299467 Norway (49%), HA-JNG04-09 Japan (27%), HS-14-12-00 Spain (22%) and H-122 Sweden (2%). The subtypes of IA were FH1 Japan (50%) and HA J04-3 Japan (50%). The substitutions of amino acid were more frequent in genotype IIIA than IA (p≤ 0.01). There was no difference in the clinical characteristics between the patients with genotype IIIA and IA. Genotype IIIA was a dominant genotype of recent HAV infection in Gwangju and Jeonnam province. This study provides valuable epidemiologic information of genetic distributions of HAV in Korea.

Coinfection of hepatitis A virus genotype IA and IIIA complicated with autoimmune hemolytic anemia, prolonged cholestasis, and false-positive immunoglobulin M anti-hepatitis E virus: a case report

A 37-year-old male presented with fever and jaundice was diagnosed as hepatitis A complicated with progressive cholestasis and severe autoimmune hemolytic anemia. He was treated with high-dose prednisolone (1.5 mg/kg), and eventually recovered. His initial serum contained genotype IA hepatitis A virus (HAV), which was subsequently replaced by genotype IIIA HAV. Moreover, at the time of development of hemolytic anemia, he became positive for immunoglobulin M (IgM) anti-hepatitis E virus (HEV). We detected HAV antigens in the liver biopsy specimen, while we detected neither HEV antigen in the liver nor HEV RNA in his serum. This is the first report of hepatitis A coinfected with two different genotypes manifesting with autoimmune hemolytic anemia, prolonged cholestasis, and false-positive IgM anti-HEV.

Longer duration of viremia and unique amino acid substitutions in a hepatitis A virus stain associated with Guillain–Barré syndrome (GBS)

The molecular characteristics of hepatitis A virus (HAV) have been studied widely though there is a paucity of data on the correlation with virological and serological findings. In the present study, the whole genome of an Indian HAV strain associated with Guillain-Barré syndrome (GBS) was characterized vis-à-vis two other Indian HAV genotype IIIA strains, associated with a self-limiting disease. The percentage nucleotide divergence displayed by the Indian strains (CP-IND, PN-IND, and GBS-IND) varied from 3 to 6, whereas the percentage amino acid divergence varied from 0.1 to 0.7 as compared to the other HAV IIIA strains (n = 5) available in the GenBank. The GBS-IND strain showed an increased rate of nonsynonymous substitutions as well as a larger number of unique and heterologous amino acid substitutions compared to the HAV IIIA GenBank strains. These amino acid substitutions in the GBS-IND strain were detected in a nonstructural protein (2C-251F) and the B-cell epitope regions of structural proteins (VP1-29E, VP1-91S, VP3-50Y, and VP4-5S). In a comparative analysis of HAV strains, homology-based models of the capsid proteins indicated a localized alteration in the surface charge distribution on the VP1 protein of GBS-IND strain and involvement of its unique amino acid substitutions in the predicted antigenic determinants. Overall, the study suggests that the unique amino acid substitutions in the GBS-IND strain may have contributed to neutralization escape of the virus leading to a longer duration of viremia.

Clinical Characterization of Hepatitis A Infection Complicated with Acute Kidney Injury and Sequence Analysis of the VP1 Region

was independently associated with AKI occurrence in non-fulminant HAV infections (odds ratios=1.094, 95% confidence interval=1.011−1.183). HAV RNA was detected in 57 (86.4%) patients: 53 strains (93.0%) showed genotype IIIA and 4 strains pre- sented genotype IA. All HAV isolates from the AKI patients belonged to the genotype IIIA and shared the identical sequences with those from the non-AKI patients. Conclusion: This study indicates that higher level of C-reactive protein is associated with AKI occurrence in non-fulminant HAV infections. There were no spe- cific nucleotide or amino acid substitutions in the VP1 region associated with AKI occurrence. (Korean J Clin Microbiol 2010;13:7-13)

Full length genomes of genotype IIIA Hepatitis A Virus strains (1995–2008) from India and estimates of the evolutionary rates and ages

With the changing epidemiology, outbreaks of Hepatitis A Virus (HAV) have been reported from different parts of India. To characterize HAV strains circulating in India (1995–2008), 6 full genome sequences of the predominant genotype, IIIA, were determined. Further, applying the Bayesian Markov Chain Monte Carlo (MCMC) framework to the full genomes of Indian HAV strains as well as other global strains (human as well as simian), we derived the mean nucleotide substitution rate and evolutionary timescales with emphasis on the age of genotype III and IIIA strains. The genomic length of all the 6 HAV isolates was 7464 nt excluding the poly A tract. Phylogenetic analysis confirmed that all the Indian isolates were close to Nor-21 ( AJ299464) and HMH ( AY644337) of subgenotype IIIA. The ORF of the isolates when compared within genotype III at amino acid level showed a highly conserved pattern. Under the best fit expansion population relaxed molecular clock model, the estimated mean substitution rate of the HAV full genomes (human and simian strains) was 1.73 × 10 −4 substitutions/site/year based on which the earliest transmission of HAV from simian to humans is estimated to have occurred about 3564 years ago. The mean substitution rate within human HAV full genomes under the same model was estimated to be 1.99 × 10 −4 substitutions/site/year. With this the mean age of genotype III strains was estimated to be 592 years while that of genotype IIIA was estimated to be 202 years. The time to the most common recent ancestor (tMRCA) of the Indian genotype IIIA isolates was calculated to be 116 years.