Gachara George, Magoro Tshifhiwa, Mavhandu Lufuno, Lum Emmaculate, Kimbi Helen K, Ndip Roland N, Bessong Pascal O
HIV/AIDS & Global Health Research Program, Department of Microbiology, University of Venda, Private Bag X5050, Thohoyandou, 0950, Limpopo, South Africa.
Department of Medical Laboratory Sciences, Kenyatta University, Nairobi, Kenya.
AIDS Res Ther. 2017 Mar 8;14(1):11. doi: 10.1186/s12981-017-0136-0.
Occult hepatitis B infection (OBI) among HIV positive patients varies widely in different geographic regions. We undertook a study to determine the prevalence of occult hepatitis B infection among HIV infected individuals visiting a health facility in South West Cameroon and characterized occult HBV strains based on sequence analyses.
Plasma samples (n = 337), which previously tested negative for hepatitis B surface antigen (HBsAg), were screened for antibodies against hepatitis B core (anti-HBc) and surface (anti-HBs) antigens followed by DNA extraction. A 366 bp region covering the overlapping surface/polymerase gene of HBV was then amplified in a nested PCR and the amplicons sequenced using Sanger sequencing. The resulting sequences were then analyzed for genotypes and for escape and drug resistance mutations.
Twenty samples were HBV DNA positive and were classified as OBI giving a prevalence of 5.9%. Out of these, 9 (45%) were anti-HBs positive, while 10 (52.6%) were anti-HBc positive. Additionally, 2 had dual anti-HBs and anti-HBc reactivity, while 6 had no detectable HBV antibodies. Out of the ten samples that were successfully sequenced, nine were classified as genotype E and one as genotype A. Three sequences possessed mutations associated with lamivudine resistance. We detected a number of mutations within the major hydrophilic region of the surface gene where most immune escape mutations occur.
Findings from this study show the presence of hepatitis B in patients without any of the HBV serological markers. Further prospective studies are required to determine the risk factors and markers of OBI.
HIV阳性患者中隐匿性乙型肝炎感染(OBI)在不同地理区域差异很大。我们开展了一项研究,以确定在喀麦隆西南部一家医疗机构就诊的HIV感染者中隐匿性乙型肝炎感染的患病率,并基于序列分析对隐匿性HBV毒株进行特征分析。
对之前乙型肝炎表面抗原(HBsAg)检测呈阴性的血浆样本(n = 337)进行乙型肝炎核心抗体(抗-HBc)和表面抗体(抗-HBs)筛查,随后进行DNA提取。然后通过巢式PCR扩增覆盖HBV表面/聚合酶重叠基因的366 bp区域,并使用桑格测序法对扩增子进行测序。接着对所得序列进行基因型分析以及逃逸和耐药突变分析。
20份样本HBV DNA呈阳性,被归类为OBI,患病率为5.9%。其中,9份(45%)抗-HBs呈阳性,10份(52.6%)抗-HBc呈阳性。此外,2份具有抗-HBs和抗-HBc双重反应性,6份未检测到HBV抗体。在成功测序的10份样本中,9份被归类为E基因型,1份为A基因型。3个序列具有与拉米夫定耐药相关的突变。我们在表面基因的主要亲水区检测到一些突变,大多数免疫逃逸突变发生在此区域。
本研究结果表明,在没有任何HBV血清学标志物的患者中存在乙型肝炎。需要进一步开展前瞻性研究以确定OBI的危险因素和标志物。