Detemmerman L, Olivier S, Bours V, Boemer F
a LaCAR MDX Technologies , Liège , Belgium.
b Biochemical Genetics Lab, Human Genetics, CHU of Liege , University of Liège , Liège , Belgium.
Hematology. 2018 Apr;23(3):181-186. doi: 10.1080/10245332.2017.1371470. Epub 2017 Sep 20.
Hemoglobin (Hb) disorders consist of thalassemia and Hb structural variants, of which the major forms are associated with severe anemia and/or vascular occlusion. Current diagnostic techniques are highly accurate and mostly based on isoelectric focusing, high-performance liquid chromatography or mass spectrometry, which often require advanced laboratory equipment. In sub-Saharan Africa, the Hb disorders are mainly associated to the pathological variants hemoglobin S (HbS) and HbC. Unfortunately, until now, it is not easy to get a diagnosis of these disorders in this area. In this study, we tested the performance of a new molecular diagnostic tests on qualified samples.
The Human Hb S/C Lamp assay is a new polymerase chain reaction test able to detect HbS, HbC and HbA alleles without DNA extraction, directly on fresh or frozen blood samples, or on dried blood spots (DBS). In this study, we compared the genotyping of 248 blood samples (56 whole blood and 192 DBS) with this LAMP assay to the routine diagnostic methods performed in the genetics lab at the university hospital of Liège.
Our results show that the LAMP method can detect HbS and HbC with an accuracy of 100%. Moreover, this test can be used for the neonatal screening because we did not observe any interference with fetal Hb.
To our knowledge, this method is the only molecular assay that can be performed directly on dried blood cards without DNA extraction, lowering handling, turnaround time and costs.
血红蛋白（Hb）疾病包括地中海贫血和Hb结构变异，其主要形式与严重贫血和/或血管闭塞有关。当前的诊断技术高度准确，且大多基于等电聚焦、高效液相色谱或质谱分析，这些技术通常需要先进的实验室设备。在撒哈拉以南非洲地区，Hb疾病主要与病理性变异血红蛋白S（HbS）和HbC有关。遗憾的是，迄今为止，在该地区对这些疾病进行诊断并不容易。在本研究中，我们对合格样本测试了一种新的分子诊断检测方法的性能。
人类Hb S/C Lamp检测法是一种新型聚合酶链反应检测方法，无需提取DNA，可直接对新鲜或冷冻血液样本或干血斑（DBS）检测HbS、HbC和HbA等位基因。在本研究中，我们将采用这种Lamp检测法对248份血液样本（56份全血和192份DBS）进行基因分型的结果，与列日大学医院遗传学实验室采用的常规诊断方法进行了比较。
我们的结果表明，Lamp方法检测HbS和HbC的准确率达100%。此外，该检测可用于新生儿筛查，因为我们未观察到其对胎儿血红蛋白有任何干扰。
据我们所知，该方法是唯一一种无需提取DNA即可直接在干血卡上进行的分子检测方法，减少了操作、周转时间和成本。
