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Molecular characterization of hepatitis A virus strains in a tertiary care health set up in north western India.

Singh MP, Majumdar M, Thapa BR, Gupta PK, Khurana J, Budhathoki B, Ratho RK - Indian J. Med. Res. (2015)

Bottom Line: Sequencing of 15 representative strains was carried out and the circulating genotype was found to be III A.The cumulative substitution in AVH strains Vs ALF strains as compared to GBM, Indian and prototype strain in the 200-500 region of 5' NTR was comparable.Our results showed hepatitis A still a disease of children with III A as a circulating genotype in this region.

View Article: PubMed Central - PubMed

Affiliation: Department of Virology, Postgraduate Institute of Medical Education & Research, Chandigarh, India.

ABSTRACT

Background & objectives: Hepatitis A virus usually causes acute viral hepatitis (AVH) in the paediatric age group with a recent shift in age distribution and disease manifestations like acute liver failure (ALF). This has been attributed to mutations in 5'non-translated region (5'NTR) which affects the viral multiplication. The present study was aimed to carry out the molecular detection and phylogenetic analysis of hepatitis A virus strains circulating in north western India.

Methods: Serum samples from in patients and those attending out patient department of Pediatric Gastroenterology in a tertiary care hospital in north India during 2007-2011 with clinically suspected AVH were tested for anti-hepatitis A virus (HAV) IgM antibodies. Acute phase serum samples were subjected to nested PCR targeting the 5'NTR region followed by sequencing of the representative strains.

Results: A total of 1334 samples were tested, 290 (21.7%) were positive for anti-HAV IgM antibody. Of these, 78 serum samples (< 7 days old) were subjected to PCR and 47.4% (37/78) samples showed the presence of HAV RNA. Children < 15 yr of age accounted for majority (94%) of cases with highest seropositivity during rainy season. Sequencing of 15 representative strains was carried out and the circulating genotype was found to be III A. The nucleotide sequences showed high homology among the strains with a variation ranging from 0.1-1 per cent over the years. An important substitution of G to A at 324 position was shown by both AVH and ALF strains. The cumulative substitution in AVH strains Vs ALF strains as compared to GBM, Indian and prototype strain in the 200-500 region of 5' NTR was comparable.

Interpretation & conclusion: Our results showed hepatitis A still a disease of children with III A as a circulating genotype in this region. The mutations at 5'NTR region warrant further analysis as these affect the structure of internal ribosomal entry site which is important for viral replication.

No MeSH data available.


Related in: MedlinePlus

Genetic relatedness of 15 representative HAV strains reported from north India from 2007-2011 with 23 reference HAV strains retrieved from GenBank. Comparison is based on 500 nucleotides from 5’NTR region and analyzed by neighbor-joining method. Bootstrap values of 1000 replicates were used for statistical verification. Our seven strains clustered with isolate from west India, one ALF sequence was closely similar to wild type of HAV IIIA from Germany, three showed close association with HAV strain from a Guillain Barre patient, another three were related to strains isolated from Norway and Japan. Year-wise representation of strains: •, 2007; blank, 2008; □, 2009; ▴, 2010; ○, 2011.
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Figure 2: Genetic relatedness of 15 representative HAV strains reported from north India from 2007-2011 with 23 reference HAV strains retrieved from GenBank. Comparison is based on 500 nucleotides from 5’NTR region and analyzed by neighbor-joining method. Bootstrap values of 1000 replicates were used for statistical verification. Our seven strains clustered with isolate from west India, one ALF sequence was closely similar to wild type of HAV IIIA from Germany, three showed close association with HAV strain from a Guillain Barre patient, another three were related to strains isolated from Norway and Japan. Year-wise representation of strains: •, 2007; blank, 2008; □, 2009; ▴, 2010; ○, 2011.

Mentions: Seven of our strains (JX481901, JX481902, JX481904, JX481906, JX481908, JX390730 and JX390731) clustered along the Indian isolate of HAV (FJ 360734.1) reported from Pune, western India. One of our ALF sequence (JX390732) showed close similarity to the wild type strain of HAV genotype IIIA reported from blood donation camp in Germany AY644337. Three of our sequences (JX481905, JX481910, and JX390729) showed close association with a reported HAV strain from a Guillain-Barre patient with longer duration of viraemia (DQ991029). Another three sequences (JX390733, JX481907, and JX481909) were related to HAV isolate (AJ299464) reported from a HAV outbreak in Norway associated with parental transmission among hemophiliacs and intravenous drug users. One of our strains (JX481903) was related to a strain isolated from Japan AB279732. (Fig. 2).


Molecular characterization of hepatitis A virus strains in a tertiary care health set up in north western India.

Singh MP, Majumdar M, Thapa BR, Gupta PK, Khurana J, Budhathoki B, Ratho RK - Indian J. Med. Res. (2015)

Genetic relatedness of 15 representative HAV strains reported from north India from 2007-2011 with 23 reference HAV strains retrieved from GenBank. Comparison is based on 500 nucleotides from 5’NTR region and analyzed by neighbor-joining method. Bootstrap values of 1000 replicates were used for statistical verification. Our seven strains clustered with isolate from west India, one ALF sequence was closely similar to wild type of HAV IIIA from Germany, three showed close association with HAV strain from a Guillain Barre patient, another three were related to strains isolated from Norway and Japan. Year-wise representation of strains: •, 2007; blank, 2008; □, 2009; ▴, 2010; ○, 2011.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4418158&req=5

Figure 2: Genetic relatedness of 15 representative HAV strains reported from north India from 2007-2011 with 23 reference HAV strains retrieved from GenBank. Comparison is based on 500 nucleotides from 5’NTR region and analyzed by neighbor-joining method. Bootstrap values of 1000 replicates were used for statistical verification. Our seven strains clustered with isolate from west India, one ALF sequence was closely similar to wild type of HAV IIIA from Germany, three showed close association with HAV strain from a Guillain Barre patient, another three were related to strains isolated from Norway and Japan. Year-wise representation of strains: •, 2007; blank, 2008; □, 2009; ▴, 2010; ○, 2011.
Mentions: Seven of our strains (JX481901, JX481902, JX481904, JX481906, JX481908, JX390730 and JX390731) clustered along the Indian isolate of HAV (FJ 360734.1) reported from Pune, western India. One of our ALF sequence (JX390732) showed close similarity to the wild type strain of HAV genotype IIIA reported from blood donation camp in Germany AY644337. Three of our sequences (JX481905, JX481910, and JX390729) showed close association with a reported HAV strain from a Guillain-Barre patient with longer duration of viraemia (DQ991029). Another three sequences (JX390733, JX481907, and JX481909) were related to HAV isolate (AJ299464) reported from a HAV outbreak in Norway associated with parental transmission among hemophiliacs and intravenous drug users. One of our strains (JX481903) was related to a strain isolated from Japan AB279732. (Fig. 2).

Bottom Line: Sequencing of 15 representative strains was carried out and the circulating genotype was found to be III A.The cumulative substitution in AVH strains Vs ALF strains as compared to GBM, Indian and prototype strain in the 200-500 region of 5' NTR was comparable.Our results showed hepatitis A still a disease of children with III A as a circulating genotype in this region.

View Article: PubMed Central - PubMed

Affiliation: Department of Virology, Postgraduate Institute of Medical Education & Research, Chandigarh, India.

ABSTRACT

Background & objectives: Hepatitis A virus usually causes acute viral hepatitis (AVH) in the paediatric age group with a recent shift in age distribution and disease manifestations like acute liver failure (ALF). This has been attributed to mutations in 5'non-translated region (5'NTR) which affects the viral multiplication. The present study was aimed to carry out the molecular detection and phylogenetic analysis of hepatitis A virus strains circulating in north western India.

Methods: Serum samples from in patients and those attending out patient department of Pediatric Gastroenterology in a tertiary care hospital in north India during 2007-2011 with clinically suspected AVH were tested for anti-hepatitis A virus (HAV) IgM antibodies. Acute phase serum samples were subjected to nested PCR targeting the 5'NTR region followed by sequencing of the representative strains.

Results: A total of 1334 samples were tested, 290 (21.7%) were positive for anti-HAV IgM antibody. Of these, 78 serum samples (< 7 days old) were subjected to PCR and 47.4% (37/78) samples showed the presence of HAV RNA. Children < 15 yr of age accounted for majority (94%) of cases with highest seropositivity during rainy season. Sequencing of 15 representative strains was carried out and the circulating genotype was found to be III A. The nucleotide sequences showed high homology among the strains with a variation ranging from 0.1-1 per cent over the years. An important substitution of G to A at 324 position was shown by both AVH and ALF strains. The cumulative substitution in AVH strains Vs ALF strains as compared to GBM, Indian and prototype strain in the 200-500 region of 5' NTR was comparable.

Interpretation & conclusion: Our results showed hepatitis A still a disease of children with III A as a circulating genotype in this region. The mutations at 5'NTR region warrant further analysis as these affect the structure of internal ribosomal entry site which is important for viral replication.

No MeSH data available.


Related in: MedlinePlus