Hepatitis B Transmission: Is Vertical Transmission the Major Route in Intermediate Endemic Areas? A Proof-of-Concept Study Based on Mother–Child Genotypes

The route of hepatitis B transmission is believed to be horizontal in India, though pediatric studies showed mother as source in the majority of chronic HBV (CHB) cases. We aimed at establishing the fact that mother–child transmission is the main route of acquisition by documenting genotypically identical viruses in mother–child pairs. Blood samples of consecutive children (≤18 years) with CHB and high DNA (>10,000 IU/mL) and their positive mother were collected from January 2013 to December 2015. Polymerase chain reaction (PCR) products of HBV-DNA were amplified and sequenced by using BigDye Terminator Cycle Sequencing Kit v3.1 and aligned with previously described sequences in the region of interest for genotypes A to G by using BioEdit software. Phylogenetic tree was generated using p-distance algorithm in MEGA software version 6. Genotyping of 59 (33 children and 26 mothers) subjects include genotype A in 24 (40.7%) and genotype D in 35 (59.3%). Both mother–child pair genotyping was possible in 25. The median age of 25 children (20 males) was 9 (interquartile range, IQR: 4–11). The distribution of genotypes among mother–child pairs was similar. The concordance between children and their mothers was 24 of 25 (96%). Evolutionary analyses showed significant similarities between mother and child sequences for both genotype A and D, suggesting thereby the same virus. In conclusion, mother–baby transmission seems to be the major route of acquisition of HBV in children in India and near-complete homology in genetic sequences between mother–child pairs is definite proof for that. However, a larger epidemiological study is required to substantiate our findings.

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Complex structural variations in non-human primate hepatitis B virus

Virology Journal ◽

10.1186/s12985-021-01667-0 ◽

2021 ◽

Vol 18 (1) ◽

Author(s):

Yoshihito Nagura ◽

Kei Fujiwara ◽

Kentaro Matsuura ◽

Etsuko Iio ◽

Yasuhito Tanaka ◽

...

Keyword(s):

Hepatitis B ◽

Core Region ◽

Structural Variations ◽

Hbv Genotype ◽

The Core ◽

B Virus ◽

Genetic Sequences ◽

Complex Structural ◽

Genotype A ◽

Human Primate

Abstract Background Recent genome sequence technology has revealed a novel type of genetic rearrangement referred to as complex structural variations (SVs). Previous studies have elucidated the complex SVs in human hepatitis B viruses (HBVs). In this study, we investigated the existence of complex SVs in HBVs from non-human primates (NHPs). Methods Searches for nucleotide sequences of NHP HBV were conducted using the PubMed, and genetic sequences were retrieved from databases. The candidate genetic sequences harboring complex SVs were analyzed using the CLUSTALW program and MAFFT. Additional bioinformatical analyses were performed to determine strains with complex SVs and to elucidate characteristics of NHP HBV strains. Results One hundred and fifty-four HBV strains from NHPs were identified from databases. SVs and complex SVs were observed in 11 (7.1%) strains. Three gibbon HBV (GiHBV) strains showed complex SVs consisting of an insertion and a deletion in the pre-S1 region. One GiHBV strain possessed a 6-nt insertion, which are normally specific to human HBV genotype A (HBV/A) in the Core region, and further analyses clarified that the 6-nt insertion was not caused by recombination, but rather by simple insertion. Another chimpanzee HBV strain showed complex SVs in the pre-S1 region, which were composed of human HBV/E, G-specific polymorphic SV, and an additional 6-nt insertion. Conclusions In this study, complex SVs were observed in HBV strains from NHPs, in addition to human HBV strains, as shown in previous studies. These data suggest that complex SVs could also be found in other members of hepadnaviruses, and may play a role in their genetic diversity.

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Identification of Tinomiscium petiolare from Vietnam using the DNA barcode

PROCEEDINGS ON APPLIED BOTANY GENETICS AND BREEDING ◽

10.30901/2227-8834-2021-2-114-122 ◽

2021 ◽

Vol 182 (2) ◽

pp. 114-122

Author(s):

B. B. Thinh ◽

R. V. Doudkin ◽

L. D. Chac ◽

H. V. Chinh ◽

Q. V. Hoi ◽

...

Keyword(s):

Dna Sequences ◽

Dna Barcode ◽

Pcr Amplification ◽

Rapid Identification ◽

Medicinal Species ◽

Cycle Sequencing ◽

Stage Of Development ◽

Pcr Products ◽

Genetic Sequences ◽

Total Dna

Background. Tinomiscium petiolare Hook.f. & Thomson is a medicinal species of the family Menispermaceae. This species is currently being intensively exploited for therapeutic purposes. Precise and rapid identification of T. petiolare is critical and essential for the classification, propagation, use and conservation of its genetic resources. In recent years, DNA barcoding has been known to be a fast and sensitive method for identifying species at any stage of development, using short DNA sequences. In this study we have performed the identification of T. petiolare specimens in Vietnam based on the sequence analysis of 4 DNA barcode loci: ITS, matK, rbcL and rpoC.Materials and methods. Total DNA was extracted from leaf samples using DNeasy Plant Mini Kit. PCR amplification of the ITS, matK, rbcL and rpoC regions was carried out on the GeneAmp PCR System 9700 with specific primers. The purified PCR products were sequenced on the ABI 3500 Genetic Analyzer system, using BigDye®Terminator v3.1 Cycle Sequencing Kit. These genetic sequences were analyzed and compared, and a phylogenetic tree was constructed using BioEdit, BLAST, and MEGA 6 programs.Results and conclusion. The success rate of amplification and sequencing was 100% for all 4 DNA barcode loci (ITS, matK, rbcL and rpoC) in the studied specimens. The produced sequence sizes of ITS, matK, rbcL and rpoC in the specimens were 574 bp, 810 bp, 527 bp and 488 bp, respectively. Further, we identified that all studied specimens were genetically related to each other and associated with the same species T. petiolare. Overall, the results of the study generated the most complete DNA barcode database of T. petiolare collected in Vietnam, contributing to the taxonomy and identification of this species.

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Detection of Hepatitis B Genotypes in HBsAg & HBeAg Carriers

Journal of Biomedical and Pharmaceutical Research ◽

10.32553/jbpr.v7i5.556 ◽

2018 ◽

Vol 7 (5) ◽

Author(s):

Salman Khan ◽

Molly Madan

Keyword(s):

Chronic Hepatitis ◽

Viral Load ◽

Hepatitis B Virus ◽

Hepatitis B ◽

Chronic Hepatitis B ◽

Central Research ◽

Elisa Kit ◽

B Virus ◽

Negative Chronic Hepatitis ◽

Genotype A

Objective:- Hepatitis B is noteworthy medical issues that may include the late continuation of liver cirrhosis and hepatocellular carcinoma. The present study aimed for the detection and diffrentiation of Hepatitis B virus HBsAg inactive non-replicative carriers, HBeAg-positive inactive replicative carriers, active carriers & HBeAg-negative chronic hepatitis B by Real Time PCR and their genotyping Methods: This research conducted on 245 positive for HBsAg, 118 (48.16 %) were male and 127 (51.84%) were female patients, which was performed in central research station labortory of Microbiology at netaji subhash Chandra Bose subharti Medical College and Hospital, Meerut Between march 2016 to November 2017 The sera were separated and screened for HBsAg by ELISA kit. Positive samples for HBsAg were tested for HBeAg ELISA kit and DNA Viral load then sequenced for genotying Results:. Of the 245 HBsAg Positive case 55 (1.12%) were HBeAg positive. In 16 PCR positive and HBV genotyping, In HBsAg inactive Non-Replicative 37.5% (n=6) genotype-B and 6.25% (n=1) genotype-A, In HBeAg inactive Replicative 12.5% (n=2) genotype-B and 12.5% (n=2) genotype-A and In HBeAg Active Chronic Hepatitis B 18.75% (n=3) genotype-B and 12.5% (n=2) genotype-A were detected Conclusions: Management strategy, using HBsAg, HBeAg and HBV DNA viral load, seems adequate for the confirmation and diffrentiation of Hepatitis B virus inactive, active carriers & HBeAg-negative chronic hepatitis B patients and genotype B was more prevalent in comparission to genotype A. Distribution of carriers & genotypes, help physicians to prescribe proper antiviral/interferon therapy according to current genotyping pattern in this region Keywords: Hepatitis B virus, Carrier State, HBsAg, HBeAg, RT-PCR

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