scholarly journals Disproportionate Distribution of HBV Genotypes A and D and the Recombinant Genotype D/E in the High and Low HBV Endemic Regions of Uganda: A Wake-Up Call for Regional Specific HBV Management

2022 ◽  
Vol 2022 ◽  
pp. 1-15
Author(s):  
Hussein Mukasa Kafeero ◽  
Dorothy Ndagire ◽  
Ponsiano Ocama ◽  
Charles Drago Kato ◽  
Eddie Wampande ◽  
...  

Background. Hepatitis B virus (HBV) is the leading cause of liver-related diseases. In Uganda, there is a regional disparity in the HBV burden. Our study was aimed at establishing the circulating genotypes in a low and a high endemic region to give plausible explanations for the differences in regional burden and guide the future management of the disease. Methods. A total of 200 HBsAg-seropositive subjects were recruited into the study by convenience sampling. The HBsAg Rapid Test Strip (Healgen Scientific Limited Liability Company, Houston, TX77047- USA) was used to screen for HBsAg while the Roche machine (Roche, Basel Switzerland/Abbot Technologies (USA)) was used to determine the viral load. The Chemistry Analyzer B120 (Mindray, China) was used for chemistry analysis. For HBV genotyping, total DNA was extracted from whole blood using the QIAamp® DNA extraction kit. Nested PCR amplification was performed using Platinum Taq DNA Polymerase (Invitrogen Corporation, USA) to amplify the 400 bp HBV polymerase gene. Purification of nested PCR products was performed using Purelink PCR product purification kit (Life Technologies, USA). Automated DNA sequencing was performed using BigDye Terminator v3.1 Cycle Sequencing Kit on 3130 Genetic Analyzer (Applied Biosystems, USA). The NCBI HBV genotyping tool (https://www.ncbi.nlm.nih.gov/projects/genotyping/formpage.cgi) was used for determination of genotype for each HBV sequence. Pearson’s chi-square, multinomial logistic regression, and Mann–Whitney U tests were used for the analysis. All the analyses were done using SPSS version 26.0 and MedCalc software version 19.1.3 at 95% CI. A p < 0.05 was considered statistically significant. Results. Majority of our study subjects were female (64.5%), youth (51.0%), and married (62.0%). Overall, genotype A was the most prevalent (46%). Genotype D and the recombinant genotype D/E were proportionately more distributed in the high endemic (38.2%) and low endemic (36.5%) regions, respectively. Genotype D was significantly more prevalent in the high endemic region and among the elderly ( p < 0.05 ). Genotype D was significantly associated with elevated viral load and direct bilirubin ( p < 0.05 ). The recombinant genotype D/E was significantly associated with elevated viral load ( p < 0.05 ). Similarly, genotype A was significantly associated with elevated AST and GGT, lowered viral load, and normal direct bilirubin levels ( p < 0.05 ). Conclusion. There is disproportionate distribution of genotypes A and D and the recombinant genotype D/E in the low and high endemic regions of Uganda. This probably could explain the differences in endemicity of HBV in our country signifying the need for regional specific HBV management and control strategies.

Author(s):  
Salman Khan ◽  
Molly Madan

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


2021 ◽  
Vol 21 (1) ◽  
Author(s):  
Xianlin Ye ◽  
Tong Li ◽  
Ran Li ◽  
Heng Liu ◽  
Junpeng Zhao ◽  
...  

Abstract Background Hepatitis B virus (HBV) infection is a major concern for blood safety in high-prevalence HBV countries such as China. In Shenzhen, dual hepatitis B surface antigen (HBsAg) enzyme-linked immunosorbent assays (ELISAs) have been adopted in parallel with nucleic acid testing (NAT) for donors for over a decade. A small proportion of blood donors test reactive (R) for HBsAg but negative through routine NAT, which can lead to HBV infection with an extremely low viral load. Objectives We aimed to investigate and analyze the molecular characteristics of HBV among blood donors that tested HBsAg R in a single ELISA test. Methods Blood donations were evaluated in this study if confirmed HBsAg R through one of two ELISA kits. Samples with non-reactive (NR) results by NAT were collected and tested for HBsAg by chemiluminescent microparticle immunoassay (CLIA) with a neutralization test. The level of HBsAg was further assessed by electrochemiluminescence immunoassay (ECLIA). The viral basic core promoter (BCP) and pre-core (PC) and S regions were amplified by nested PCR. Quantitative real-time PCR (qPCR) for viral load determination and individual donation (ID)-NAT were adopted simultaneously. HBsAg was confirmed with CLIA, ECLIA, nested PCR, qPCR, and ID-NAT. Results Of the 100,252 donations, 38 and 41 were identified as HBsAg R with Wantai and DiaSorin ELISA kits, respectively. Seventy-nine (0.077%, 79/100,252) blood samples with ELISA R-NR and NAT NR results were enrolled in the study. Of these, 17 (21.5%,17/79) were confirmed as HBsAg-positive. Of the 14 genotyped cases, 78.6% (11/14) were genotype B, and C and D were observed in two and one sample, respectively. Mutations were found in the S gene, including Y100C, Y103I, G145R, and L175S, which can affect the detection of HBsAg. A high-frequency mutation, T1719G (93.3%), was detected in the BCP/PC region, which reduced the viral replication. Conclusion A small number of blood samples with HBsAg ELISA R-NR and NAT NR results were confirmed as HBV infection, viral nucleic acids were found in most of the samples through routine NAT methods. It is necessary to employ more sensitive and specific assays for the detection of HBV infection among blood donors.


Author(s):  
Ujjal Poddar ◽  
Mercilena Benjamin ◽  
Rakesh Aggarwal ◽  
Aditya Narayan Sarangi ◽  
Amrita Mathias ◽  
...  

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.


2014 ◽  
Vol 35 (2) ◽  
pp. 203-224 ◽  
Author(s):  
Torben Riehl ◽  
Nils Brenke ◽  
Saskia Brix ◽  
Amy Driskell ◽  
Stefanie Kaiser ◽  
...  

AbstractField and laboratory protocols that originally led to the success of published studies have previously been only briefly laid out in the methods sections of scientific publications. For the sake of repeatability, we regard the details of the methodology that allowed broad-range DNA studies on deep-sea isopods too valuable to be neglected. Here, a comprehensive summary of protocols for the retrieval of the samples, fixation on board research vessels, PCR amplification and cycle sequencing of altogether six loci (three mitochondrial and three nuclear) is provided. These were adapted from previous protocols and developed especially for asellote Isopoda from deep-sea samples but have been successfully used in some other peracarids as well. In total, about 2300 specimens of isopods, 100 amphipods and 300 tanaids were sequenced mainly for COI and 16S and partly for the other markers. Although we did not set up an experimental design, we were able to analyze amplification and sequencing success of different methods on 16S and compare success rates for COI and 16S. The primer pair 16S SF/SR was generally reliable and led to better results than universal primers in all studied Janiroidea, except Munnopsidae and Dendrotionidae. The widely applied universal primers for the barcoding region of COI are problematic to use in deep-sea isopods with a success rate of 45–79% varying with family. To improve this, we recommend the development of taxon-specific primers.


2009 ◽  
Vol 29 (6) ◽  
pp. 469-473 ◽  
Author(s):  
Edna Maria Cavallini Sanches ◽  
Susi M. Pacheco ◽  
Alison S. Cericatto ◽  
Rosane M. Melo ◽  
Edson Molleta Colodel ◽  
...  

Pneumocystis has been isolated from a wide range of unrelated mammalian hosts, including humans, domestic and wild animals. It has been demonstrated that the genome of Pneumocystis of one host differs markedly from that of other hosts. Also, variation in the chromosome and DNA sequence of Pneumocystis within a single host species has been observed. Since information about the occurrence and nature of infections in wild animals is still limited, the objective of this work was to detect the presence of Pneumocystis sp. in lungs of bats from two states from Brazil by Nested-PCR amplification. The bats, captured in caves and in urban areas, were obtained from the Program of Rabies Control of two States in Brazil, Mato Grosso and Rio Grande do Sul, located in the Mid-Western and Southern regions of the country, respectively. DNAs were extracted from 102 lung tissues and screened for Pneumocystis by nested PCR at the mtLSU rRNA gene and small subunit of mitochondrial ribosomal RNA (mtSSU rRNA). Gene amplification was performed using the mtLSU rRNA, the primer set pAZ102H - pAZ102E and pAZ102X - pAZY, and the mtSSU rRNA primer set pAZ102 10FRI - pAZ102 10R-RI and pAZ102 13RI - pAZ102 14RI. The most frequent bats were Tadarida brasiliensis (25), Desmodus rotundus (20), and Nyctinomops laticaudatus (19). Pneumocystis was more prevalent in the species Nyctinomops laticaudatus (26.3% = 5/19), Tadarida brasiliensis (24% = 6/25), and Desmodus rotundus (20% = 4/20). Besides these species, Pneumocystis also was detected in lungs from Molossus molossus (1/11, 9.1%), Artibeus fimbriatus (1/1, 100%), Sturnira lilium (1/3, 33.3%), Myotis levis (2/3, 66.7%)and Diphylla ecaudata (1/2, 50%). PCR products which could indicate the presence of Pneumocystis (21.56%) were identified in DNA samples obtained from 8 out of 16 classified species from both states (5 bats were not identified). This is the first report of detection of Pneumocystis in bats from Brazil.


2015 ◽  
Vol 54 (2) ◽  
pp. 162-168 ◽  
Author(s):  
Sha Lu ◽  
Xiqing Li ◽  
Richard Calderone ◽  
Jing Zhang ◽  
Jianchi Ma ◽  
...  

2013 ◽  
Vol 7 (07) ◽  
pp. 541-549
Author(s):  
Myo Nyein Aung ◽  
Wattana Leowattana ◽  
Khine Nwe Win ◽  
Noppadon Tangpukdee ◽  
Sant Muangnoicharoen

Introduction: Chronic hepatitis B (CHB) is a globally common infectious disease. Its clinical course is complicated. In Southeast Asia, nucleos(t)ide analogues (NA) are commonly used drugs for CHB treatment. Composite treatment outcome has often been used in CHB clinical practice, but rarely predicted epidemiologically. This study aimed to compare the composite treatment outcome between CHB patients with low and high treatment-naïve viral load, and to identify its predictors Methodology: This retrospective cohort study followed up 95 CHB patients on NA treatment for a year. Composite treatment outcome was defined as undetectable HBV DNA level, ALT normalization and, HBeAg clearance in the case of HBeAg-positive patients. Multinomial logistic regression analysis was applied to analyze the significant treatment response predictors. Results: Complete composite treatment outcome was achieved by 52% of CHB patients with an initial viral load < 6.5 log 10 copies /ml, but 31% of those had an initial viral load ≥ log 6.5 log 10 copies /ml. Outcome was predicted by HBeAg negativity (adjusted relative risk ratio, aRRR = 11.1, 95 % confidence interval, CI 3-41.3) and ALT normalization within the sixth month of therapy (aRRR = 6.7, CI 1.8-24.9). An elevation of ALT to more than 1.5 times the normal value (40 IU/ml) can lead to an incomplete response on NA therapy (aRRR = 6.2, CI 1.5-26.6.) Conclusion: Routine clinical markers other than pre-treatment viral load predicted composite CHB outcome on NA Therapy.


2009 ◽  
Vol 133 (1-2) ◽  
pp. 34-42 ◽  
Author(s):  
James F.X. Wellehan ◽  
April L. Childress ◽  
Rachel E. Marschang ◽  
April J. Johnson ◽  
Elaine W. Lamirande ◽  
...  
Keyword(s):  

2009 ◽  
Vol 3 (07) ◽  
pp. 511-516 ◽  
Author(s):  
Vikas Agashe ◽  
Shubhada Shenai ◽  
Ganesh Mohrir ◽  
Minal Deshmukh ◽  
Anita Bhaduri ◽  
...  

Background: We conducted a study of osteoarticular tuberculosis in patients from private and public settings in a disease endemic area. Our objective was to assess the role of mycobacterial culture and polymerase chain reaction (PCR) in the diagnosis of osteoarticular tuberculosis (TB) in settings where only clinical and imaging diagnosis form the basis for treatment. Methodology: Ninety-three consecutive specimens collected from clinically suspected patients of osteoarticular TB were screened for bacterial culture, mycobacterial culture and in-house nested PCR. In addition, specimens were examined by imaging and histopathology. Ten specimens collected from patients suffering from other bone diseases were included as negative controls. Results: Of the 93 clinically suspected TB patients, mycobacterial culture was positive for Mycobacterium tuberculosis (MTB) in 47 (51%) patients who were confirmed as definite TB cases. Of the remaining patients, 16 (17%) were diagnosed as probable, 19 (20%) as possible, and 11 (12%) as only clinically suspected TB cases. In-house nested PCR was positive in 65 (70%) cases. Fifteen patients were resistant to one or more anti-tuberculous drugs; twelve patients were multi-drug resistant, two of whom were extensively drug resistant. Conclusion: Mycobacterial cultures using liquid media with susceptibility should form the backbone of management of osteoarticular TB. Nested PCR enhances the sensitivity if performed in addition to culture.


2018 ◽  
Vol 68 (1) ◽  
pp. 27 ◽  
Author(s):  
A. GIANNAKOPOULOS ◽  
C. N. TSOKANA ◽  
E. PAPADOPOULOS ◽  
V. SPYROU ◽  
D. C. CHATZOPOULOS ◽  
...  

In this study we aimed to determine the prevalence of Leishmania spp infection and to report the geographical distribution of Leishmania spp infected cats from Thessaly, central Greece, an endemic region for canine and human leishmaniosis. Blood samples were collected from a total of 150 cats (34 stray and 116 owned) from the area of Thessaly, i.e. Larissa (n=6), Volos (n=94), Trikala (n=30) and Karditsa (n=20). Data on signalment and living conditions were registered for each cat. The samples were tested by Internal Transcribed Spacer 1 nested PCR (ITS1 nested PCR). Geographical Information System (GIS) was used to define the geographical distribution of the Leishmania spp infected cats in relation to the land uses and the altitude. In total, 13.3% (95% CI: 8.3-19.8) of the sampled cats were found positive for Leishmania spp DNA. More precisely, the results are 12.8% (95% CI 6.8-21.2) in Volos, 20% (95% CI 6.8-21.2) in Trikala and 10% (95% CI 12-31.7) in Karditsa while none of the six cats examined from the region of Larissa was found PCR positive. The Leishmania spp infected cats were found in artificial surfaces and associated areas and in cultivated and managed areas with a mean altitude of 81,7 m above sea level (range 14 - 225 ± 51.57 SD). No significant correlation was found between Leishmania spp infection and signalment or living conditions, thus suggest ing that the infection is uniformly distributed among the screened feline population. The findings of this study along with the occurrence of competent vectors of Leishmania infantum in Thessaly, support the hypothesis that cats may serve as a reservoir for Leishmania spp in this region and highlight the need for surveillance in feline population and implementation of control measures. However, the epidemiological role of cats in the maintenance and transmission of Leishmania spp has yet to be defined and deserves further investigation.


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