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RISS 인기검색어
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- 저자 : ( Shah Khushbu ) (검색결과 1 건)
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Distribution of Hepatitis B Genotypes Among Hepatitis B Patients in Central Nepal
( Gupta Kumar Santosh ),( Shah Khushbu ) 대한간학회 2020 춘·추계 학술대회 (KASL) Vol.2020 No.1
Aims: To find the distribution of hepatitis B genotypes among hepatitis B infected patients in Central Nepal. To determine the HBeAg, HBeBa and viral load among HBsAg positive patients, to determine HBV genotypes and to correlate it with clinical problems. Methods: This was a prospective type of study. The research was conducted in Decode Genomics and Research Center, Sinamangal, Kathmandu, Nepal. Samples were taken from HBsAg positive patients visiting DGRC within the Nepal. 106 samples was taken for this study. The basic criteria for this study were separated as Inclusion criteria and Exclusion criteria. The Inclusion criteria was only HBsAg positive cases. And, the Exclusion criteria were HIV positive cases, HBsAg negative cases and International patients. The equipment and reagents that were required in this study are listed in the ANNEX I. 3ml Blood sample had been collected on EDTA vial by following the standard procedure of blood collection. The viral DNA from blood samples had been extracted using SpinstarTM Viral Nucleic Acid Extraction kits following manufacturer protocol. The detailed protocol is included in ANNEX III. Viral load estimation on each blood samples had been done using RealStar® HBV PCR kit. Further details is included in ANNEX IV. The Hepatitis B genotype had been determined by PCR using type specific primers (Appendix II). And, the PCR product for the Genotyping had been analyzed in 2% Agarose gel using standard protocol. Results: This study was conducted at Decode Genomics and Research Centre, Sinamangal, Kathmandu, Nepal. A total of 106 HBsAg positive serum samples were collected from patients visiting DGRC, in between June 2017 to November 2017, for Viral load Testing. Written consent was taken from each patient before sample collection. Out of 106 patients enrolled for this study 68 were males and 38 were females. Age of the enrolled patients was from 13 year to 78 years. Patients enrolled for the study were from different parts of Nepal visiting DGRC for treatment of active HBV infection. DNA was extracted using SpinStarTM Viral Nucleic Acid Kit 1.0. Viral load estimation were performed using RealStar® HBV PCR kit 1.0 and genotyping by the Nested PCR using type specific primers for each genotypes. Genotype D was found predominantly followed by C. The highest viral load in our study was >10 million i.e.1936410339 IU/ml which had genotype A whereas lowest viral load was 31 IU/ml had genotype C. Out of 106 patients 42 contains high viral load (>20000). Genotype C is predominant among high viral load patients followed by unknown, A and D. 30 patients contains moderate viral load (2000-20000) and low viral load was observed in 34 patients (<2000). Among moderate viral load patients genotype D was dominant followed by unknown and C while genotype D is more common among low viral load patients. Type-specific primers were used for HBV genotyping using nested PCR. Type specific primers were designed for the detection of conserved region of genotypes A, B, C, D, E and F. Out of 106 patients enrolled for the study HBV Genotypes was determined in 59 patients while genotypes were not determined in 47 patients. Among 59 patients genotype D was present in highest number (18 patients) followed by C, A, B and Genotype F was observed only in one patients. Genotype E was not found in any patients enrolled for the study. 16 patients were found to be containing recombinant Genotypes. A total of 6 types of recombinants were seen on our study. They are B/C, B/C/D, C/D, A/C, C/F and D/A. C/D (7) recombinant was predominant followed by A/C (3), B/C/D (2) and D/A (2). B/C and C/F recombinant was seen in 1 patient each. Conclusions: Out of 106 patients enrolled for the study, HBV Genotypes was determined in 59 patients while genotypes were not determined in 47 patients. Among 59 patients genotype D was present in highest number (18 patients) followed by C, A, B and Genotype F was observed only in one patients. Genotype E was not found in any patients enrolled for the study. 16 patients were found to be containing recombinant Genotypes. A total of 6 types of recombinants were seen on our study. They are B/C, B/C/D, C/D, A/C, C/F and D/A. C/D (7) recombinant was predominant followed by A/C(3), B/C/D (2) and D/A (2). B/C and C/F recombinant was seen in 1 patient each.
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