Waheed Usman, Noor Farooq Ahmed, Saba Noore, Wazeer Akhlaaq, Qasim Zahida, Arshad Muhammad, Karimi Saira, Farooq Ahmad, Usman Javaid, Zaheer Hasan Abbas
Department of Pathology and Transfusion Medicine, Shaheed Zulfiqar Ali Bhutto Medical University, Islamabad, Pakistan.
Islamabad Blood Transfusion Authority, Ministry of National Health Services, Government of Pakistan, Islamabad, Pakistan.
J Lab Physicians. 2020 Aug;12(2):92-97. doi: 10.1055/s-0040-1716593. Epub 2020 Sep 2.
The serological testing of human immunodeficiency virus (HIV) is mandatory under the blood safety legislation of Pakistan; hence, data exist on the prevalence of HIV in blood donors. However, little is known about the molecular epidemiology of HIV in the blood donor population. Therefore, the current study was designed to study the genetic diversity of HIV-1 infection in a population of apparently healthy treatment-naive blood donors in Islamabad, Pakistan. A total of 85,736 blood donors were tested for HIV by the chemiluminescence immunoassay. All positive donor samples were analyzed for the presence of various HIV genotypes (types and subtypes). Viral ribonucleic acid was extracted from blood samples of HIV positive donors and reverse transcribed into complementary deoxyribonucleic acid (cDNA). The cDNA of all positive donors was then analyzed for the presence of various HIV genotypes (types and subtypes) by employing subtype-specific primers in a nested polymerase chain reaction. The amplified products were run on ethidium bromide-stained 2% agarose gel and visualized using a ultraviolet transilluminator. A particular subtype was assigned to a sample if the subtype-specific reaction made a band 20% highly intense compared with the band made by the subtype-independent reaction. A total of 85,736 blood donors were screened for the presence of antibodies to HIV. Out of them, 114 were initially found reactive for HIV. The repeat testing resulted in 112 (0.13%) positive donors, 95% confidence interval 0.0014 (0.0011-0.0018). These 112 samples were analyzed for molecular typing of HIV-1. The predominant HIV-1 subtype was A ( = 101) (90.1%) followed by subtype B ( = 11) (9.9%). These findings are key to understand the diversified HIV epidemic at the molecular level and should assist public health workers in implementing measures to lessen the further dissemination of these viruses in the country.
根据巴基斯坦血液安全立法,对人类免疫缺陷病毒(HIV)进行血清学检测是强制性的;因此,存在关于献血者中HIV流行率的数据。然而,对于献血者群体中HIV的分子流行病学知之甚少。因此,本研究旨在研究巴基斯坦伊斯兰堡一群明显健康、未接受过治疗的献血者中HIV-1感染的基因多样性。
共有85736名献血者通过化学发光免疫分析法进行了HIV检测。对所有阳性献血者样本分析各种HIV基因型(类型和亚型)的存在情况。从HIV阳性献血者的血样中提取病毒核糖核酸,并逆转录为互补脱氧核糖核酸(cDNA)。然后,通过在巢式聚合酶链反应中使用亚型特异性引物,对所有阳性献血者的cDNA分析各种HIV基因型(类型和亚型)的存在情况。扩增产物在溴化乙锭染色的2%琼脂糖凝胶上进行电泳,并使用紫外线透射仪进行观察。如果亚型特异性反应产生的条带强度比非亚型特异性反应产生的条带高20%,则将特定亚型指定给一个样本。
共筛查了85736名献血者是否存在HIV抗体。其中,最初发现114人对HIV反应呈阳性。重复检测结果为112名(0.13%)阳性献血者,95%置信区间为0.0014(0.0011 - 0.0018)。对这112个样本进行了HIV-1分子分型分析。主要的HIV-1亚型是A(n = 101)(90.1%),其次是B亚型(n = 11)(9.9%)。
这些发现是在分子水平上理解多样化HIV流行情况的关键,应有助于公共卫生工作者采取措施减少这些病毒在该国的进一步传播。