Demir Tülin, Yıldıran Dilara, Kılıç Selçuk
Ministry of Health, General Directorate of Public Health, Department of Microbiology Reference Laboratories and Biological Products, National HIV-AIDS and Viral Hepatitis Reference Laboratory, Ankara, Turkey.
Health Sciences University, Gülhane Defense Health Sciences Institute, Department of Medical CBRN, Ankara, Turkey.
Mikrobiyol Bul. 2021 Jan;55(1):17-29. doi: 10.5578/mb.20028.
Shortly after the first detection of human immundeficiency virus (HIV) infection in USA in 1981, the number of cases have increased gradually from all around the world. Turkey's high capacity for tourism and the unique geographic location extending between Europe and Asia, provides convenience for the passage of individuals across the countries and sexually transmitted infections including HIV, as well. According to the official data of the Ministry of Health; there are 25809 HIV positive and 1958 AIDS cases as of November 30, 2020, after the epidemic started in 1985 in Turkey. Despite the decrease in the number of newly detected HIV cases as a result of serious measures taken for the transmission of infection worldwide, the increase in the number of cases still continues in our country. Shortening the reporting period and starting treatment as soon as possible in the diagnosis of infection is critical for the control of the epidemic. For this purpose, Centers for Disease Control and Prevention (CDC) published a new test algorithm in 2010, which suggested the use of the Geenius™ HIV ½ supplemental assay test instead of western blot tests, which have been used for many years to verify HIV screening test positivity. In this study, we aimed to report the experience of the National HIV-Acquiner Immundeficiency Syndrome(AIDS) and Viral Hepatitis Reference Laboratories of Turkey in the first year of transition to the new HIV algorithm and to evaluate the diagnostic performance of Geenius™ HIV ó and line immunassay (LIA) s. A total of 2090 anti-HIV positive patient sera sent to National HIV-AIDS and Viral Hepatitis Reference Laboratories of Turkey, Ankara for HIV confirmation were included in the study. All samples were retested with a fourth-generation enzyme linked immunosorbent assay (ELISA) test (VIDAS® HIV-1/2 Duo Ultra assay, BioMerieux, France) followed by the confirmatory tests; Geenius™ HIV 1/2 confirmatory assay (BioRad, Redmond, WA) and Line-immunoassay (INNO-LIA HIV ½ Score, Fujirebio, reverse transcriptase polymerase chain reaction (RT-PCR) (artus HI Virus-1 RT-PCR, Qiagen, Germany) test and in-house HIV-2 RNA and proviral DNA PCR. The sensitivity, specificity, and the agreement of the each assay were compared. Cohen's Kappa analysis was used for the evaluation of the agreement between the tests. According to the new algorithm which recommended Geenius™ test besides HIV-1 RNA test, 1707 (81.7%) HIV-1 positive samples were identified. Of these samples; 95.9% and 95.02% were identified as HIV-1 positive by GeeniusTM and INNO-LIA, respectively. However, 2.5% of the positive samples were negative with Geenius™ and 3.5% with INNO-LIA. One and a half percentage (1.5%) of these samples were detected with Geenius™ and 1.4% with INNO-LIA as indeterminant. When all the positive samples determined with ELISA were evaluated; it was detected that,1.3% were indeterminate by Geenius™ test and 2.4% by the INNO-LIA test. When the INNO-LIA test was regarded as the gold standard method; sensitivity, specificity, positive predictive and negative predictive values of the Geenius™ test were as follows; 99.7%, 96.1%, 98.9%, and 99.1%. The agreement between INNO-LIA and Geenius™ tests was found to be 98.95% (κ= 0.969; very good). When the Geenius™ and HIV-1 PCR tests were evaluated together for the confirmation; the sensitivities of Geenius™ and INNO-LIA tests were 99.8% and 98.3%, specificities were 89.8% and 85.3%, respectively. Slight positive bands were detected in the gp36 or gp140 bands, the HIV-2 specific envelope proteins, were detected in seven samples, However, the positivity disappeared after the dilution of the samples and it was accepted as false positivite reaction due to the absence of HIV-2 RNA and proviral DNA in these samples. In conclusion; we concluded that Geenius ™ and INNO-LIA tests have a perfect agreement in HIV diagnosis and due to the rapid and reliable results provided for the HIV test protocol, Geenius™ test can be used safely as an alternative to the immunoblot tests. HIV-1 RNA testing must be performed in all HIV confirmation centers in order to detect acute HIV cases in the fast and early period which are the main reason for the updates in HIV diagnosis.
1981年美国首次检测到人类免疫缺陷病毒(HIV)感染后不久,全球范围内的病例数逐渐增加。土耳其旅游业发达,且地理位置独特,横跨欧亚大陆,为人员跨国流动提供了便利,也使得包括HIV在内的性传播感染风险增加。根据土耳其卫生部的官方数据,自1985年该国出现疫情以来,截至2020年11月30日,共有25809例HIV阳性病例和1958例艾滋病病例。尽管全球采取了严格措施控制感染传播,新检测出的HIV病例数有所下降,但我国的病例数仍在持续增加。缩短报告周期并在感染诊断后尽快开始治疗对于控制疫情至关重要。为此,美国疾病控制与预防中心(CDC)于2010年发布了一种新的检测算法,建议使用Geenius™ HIV ½补充检测法替代多年来用于验证HIV筛查检测阳性结果的蛋白印迹法。在本研究中,我们旨在报告土耳其国家HIV-获得性免疫缺陷综合征(AIDS)和病毒性肝炎参考实验室在采用新的HIV检测算法第一年的经验,并评估Geenius™ HIV ½和线性免疫测定(LIA)的诊断性能。本研究纳入了总共2090份送至土耳其安卡拉国家HIV-艾滋病和病毒性肝炎参考实验室进行HIV确认的抗HIV阳性患者血清。所有样本首先用第四代酶联免疫吸附测定(ELISA)检测(VIDAS® HIV-1/2 Duo Ultra检测法,法国生物梅里埃公司)进行复测,随后进行确认性检测:Geenius™ HIV 1/2确认检测(美国伯乐公司,华盛顿州雷德蒙德)、线性免疫测定(INNO-LIA HIV ½ Score,富士瑞必欧公司)、逆转录聚合酶链反应(RT-PCR)(artus HI Virus-1 RT-PCR,德国Qiagen公司)检测以及内部HIV-2 RNA和前病毒DNA PCR检测。比较了每种检测方法的敏感性、特异性和一致性。采用Cohen's Kappa分析评估检测方法之间的一致性。根据除HIV-1 RNA检测外还推荐使用Geenius™检测的新算法,共鉴定出1707例(81.7%)HIV-1阳性样本。在这些样本中,分别有95.9%和95.02%的样本通过Geenius™和INNO-LIA检测被鉴定为HIV-1阳性。然而,分别有2.5%和3.5%的阳性样本通过Geenius™和INNO-LIA检测结果为阴性。这些样本中分别有1.5%和1.4%的样本通过Geenius™和INNO-LIA检测结果为不确定。当评估所有通过ELISA检测确定为阳性的样本时,发现分别有1.3%和2.4%的样本通过Geenius™检测和INNO-LIA检测结果为不确定。当将INNO-LIA检测视为金标准方法时,Geenius™检测的敏感性、特异性、阳性预测值和阴性预测值如下:99.7%、96.1%、98.9%和99.1%。发现INNO-LIA和Geenius™检测之间的一致性为98.95%(κ = 0.9