Molecular analysis of HBV genotypes and subgenotypes in the Central-East region of Tunisia

<p>Abstract</p> <p>Background</p> <p>In Tunisia, country of intermediate endemicity for Hepatitis B virus (HBV) infection, most molecular studies on the virus have been carried out in the North of the country and little is known about other regions. The aim of this study was to determine HBV genotype and subgenotypes in Central-East Tunisia. A total of 217 HBs antigen positive patients were enrolled and determination of genotype was investigated in 130 patients with detectable HBV DNA. HBV genotyping methods were: PCR-RFLP on the pre-S region, a PCR using type-specific primers in the S region (TSP-PCR) and partial sequencing in the pre-S region.</p> <p>Results</p> <p>Three genotypes (D, B and A) were detected by the PCR-RFLP method and two (D and A) with the TSP-PCR method, the concordance between the two methods was 93%. Sequencing and phylogenetic analysis of 32 strains, retrieved the same genotype (D and A) for samples with concordant results and genotype D for samples with discordant results. The sequences of discordant genotypes had a restriction site in the pre-S gene which led to erroneous result by the PCR-RFLP method. Thus, prevalence of genotype D and A was 96% and 4%, respectively. Phylogenetic analysis showed the predominance of two subgenotypes D1 (55%) and D7 (41%). Only one strain clustered with D3 subgenotype (3%).</p> <p>Conclusions</p> <p>Predominance of subgenotype D7 appears to occur in northern regions of Africa with transition to subgenotype D1 in the East of the continent. HBV genetic variability may lead to wrong results in rapid genotyping methods and sequence analysis is needed to clarify atypical results.</p

Human leukocyte antigen (HLA-F) polymorphism is associated with chronic HBV infection.

Human leukocyte antigen (HLA)-F has been involved in immune regulation of infectious diseases. However, the role of HLA-F polymorphisms in hepatitis B infection outcomes remains unclear. Here, we aimed to determine HLA-F polymorphism implication in chronic HBV. Genotype analysis was performed for three single nucleotide polymorphisms (SNPs) of HLA-F and one SNP of HLA-E using PCR-SSP, in 252 Tunisian patients with chronic HBV infection stratified according to their HBV DNA levels (140 patients with low HBV DNA levels < 2000 IU/mL and 112 patients with high HBV DNA levels ≥ 2000 IU/mL) and 240 healthy controls (CTRL). The three HLA-F SNPs (HLA-F*01:02, -F*01:03 and -F*01:04) have the same allelic and genotypic frequencies in patients and in CTRL. We reported a low HLA-F*01:02 and F*01:04 allelic frequencies in the Tunisian population; however, high HLA-F*01:03 allele frequencies were observed (17%). A significant association was found between the HLA-F*01:03 allele and decreased level of HBV DNA (P = 0.02 OR 0.56, 95% CI 0.35-0.92). No significant differences were observed in haplotype distribution between patients and CTRL. A significant association of HLA-F*01:03 with the level of HBV DNA suggests an important role of HLA-F in HBV replication control

Epidemiology and clinical profile of pathogens responsible for the hospitalization of children in Sousse area, Tunisia

<div><p>This study aimed to identify a broad spectrum of respiratory pathogens from hospitalized and not-preselected children with acute respiratory tract infections in the Farhat Hached University-hospital of Sousse, Tunisia. Between September 2013 and December 2014, samples from 372 children aged between 1 month and 5 years were collected, and tested using multiplex real-time RT-PCR by a commercial assay for 21 respiratory pathogens. In addition, samples were screened for the presence of <i>Streptococcus pneumoniae</i> 16S rDNA using real-time PCR. The viral distribution and its association with clinical symptoms were statistically analyzed. Viral pathogens were detected in 342 (91.93%) of the samples of which 28.76% were single positive and 63.17% had multiple infections. The most frequent detected viruses were rhinovirus (55.64%), respiratory syncytial virus A/B (33.06%), adenovirus (25.00%), coronavirus NL63, HKU1, OC43, and 229E (21.50%), and metapneumovirus A/B (16.12%). Children in the youngest age group (1–3 months) exhibited the highest frequencies of infection. Related to their frequency of detection, RSV A/B was the most associated pathogen with patient’s demographic situation and clinical manifestations (p<0.05). Parainfluenza virus 1–4 and parechovirus were found to increase the risk of death (p<0.05). Adenovirus was statistically associated to the manifestation of gastroenteritis (p = 0.004). Rhinovirus infection increases the duration of oxygen support (p = 0.042). Coronavirus group was statistically associated with the manifestation of bronchiolitis (p = 0.009) and laryngitis (p = 0.017). <i>Streptococcus pneumoniae</i> DNA was detected in 143 (38.44%) of tested samples. However, only 53 samples had a concentration of C-reactive protein from equal to higher than 20 milligrams per liter, and 6 of them were single positive for <i>Streptocuccus pneumoniae</i>. This study confirms the high incidence of respiratory viruses in children hospitalized for acute respiratory tract infections in the Sousse area, Tunisia.</p></div

Impact of individual viral infection on patient’s clinical data and demographic information^a.

<p>Impact of individual viral infection on patient’s clinical data and demographic information<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0188325#t006fn001" target="_blank">^a</a>.</p

Characteristics of <i>S</i>. <i>pneumoniae</i> infected subjects in relation to viral infection (negative, single and multiple) and CRP concentrations^a.

<p>Characteristics of <i>S</i>. <i>pneumoniae</i> infected subjects in relation to viral infection (negative, single and multiple) and CRP concentrations<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0188325#t003fn001" target="_blank">^a</a>.</p

Summary of the statistical comparisons between positive viral infection and patient’s medical history, demographic data and clinical manifestations.

<p>Summary of the statistical comparisons between positive viral infection and patient’s medical history, demographic data and clinical manifestations.</p

Summary of the statistical associations comparing single and multiple viral infections with patient’s data.

<p>Summary of the statistical associations comparing single and multiple viral infections with patient’s data.</p