De Spiegelaere Ward, Philippé Jan, Vervisch Karen, Verhofstede Chris, Malatinkova Eva, Kiselinova Maja, Trypsteen Wim, Bonczkowski Pawel, Vogelaers Dirk, Callens Steven, Ruelle Jean, Kabeya Kabamba, De Wit Stephane, Van Acker Petra, Van Sandt Vicky, Emonds Marie-Paule, Coucke Paul, Sermijn Erica, Vandekerckhove Linos
Ghent University, Department of Internal Medicine, Ghent, Belgium.
Ghent University, Department of Clinical Chemistry, Microbiology and Immunology, Ghent, Belgium.
PLoS One. 2015 Apr 15;10(4):e0123525. doi: 10.1371/journal.pone.0123525. eCollection 2015.
Abacavir is a nucleoside reverse transcriptase inhibitor used as part of combination antiretroviral therapy in HIV-1-infected patients. Because this drug can cause a hypersensitivity reaction that is correlated with the presence of the HLA-B57:01 allotype, screening for the presence of HLA-B57:01 is recommended before abacavir initiation. Different genetic assays have been developed for HLA-B57:01 screening, each with specific sensitivity, turnaround time and assay costs. Here, a new real-time PCR (qPCR) based analysis is described and compared to sequence specific primer PCR with capillary electrophoresis (SSP PCR CE) on 149 patient-derived samples, using sequence specific oligonucleotide hybridization combined with high resolution SSP PCR as gold standard. In addition to these PCR based methods, a complementary approach was developed using flow cytometry with an HLA-B17 specific monoclonal antibody as a pre-screening assay to diminish the number of samples for genetic testing. All three assays had a maximum sensitivity of >99. However, differences in specificity were recorded, i.e. 84.3%, 97.2% and >99% for flow cytometry, qPCR and SSP PCR CE respectively. Our data indicate that the most specific and sensitive of the compared methods is the SSP PCR CE. Flow cytometry pre-screening can substantially decrease the number of genetic tests for HLA-B57:01 typing in a clinical setting.
阿巴卡韦是一种核苷类逆转录酶抑制剂，用于人类免疫缺陷病毒1型（HIV-1）感染患者的联合抗逆转录病毒治疗。由于该药物可引起与HLA-B57:01等位基因存在相关的超敏反应，因此建议在开始使用阿巴卡韦之前筛查HLA-B57:01的存在情况。已开发出不同的基因检测方法用于HLA-B57:01筛查，每种方法都有特定的灵敏度、周转时间和检测成本。在此，描述了一种基于实时荧光定量聚合酶链反应（qPCR）的新分析方法，并将其与149份患者来源样本上的序列特异性引物聚合酶链反应结合毛细管电泳（SSP PCR CE）进行比较，使用序列特异性寡核苷酸杂交结合高分辨率SSP PCR作为金标准。除了这些基于PCR的方法外，还开发了一种互补方法，使用流式细胞术和HLA-B17特异性单克隆抗体作为预筛查检测，以减少基因检测的样本数量。所有三种检测方法的最大灵敏度均>99%。然而，记录到特异性存在差异，即流式细胞术、qPCR和SSP PCR CE的特异性分别为84.3%、97.2%和>99%。我们的数据表明，在比较的方法中，最特异和敏感的是SSP PCR CE。在临床环境中，流式细胞术预筛查可大幅减少HLA-B57:01分型的基因检测数量。
