Department of Medical Genetics, School of Basic Medicine Science, Southern Medical University, Guangdong, China.
Experimental Education and Administration Center, School of Basic Medical Science, Southern Medical University, Guangdong, China.
Prenat Diagn. 2024 Oct;44(11):1310-1317. doi: 10.1002/pd.6661. Epub 2024 Sep 10.
Thalassemia is a Mendelian-inherited blood disorder with severe consequences, including disability and mortality, making it a significant public health concern. Therefore, there is an urgent need for precise diagnostic technologies. We introduce two innovative diagnostic techniques for thalassemia, SNPscan and CNVplex, designed to enhance molecular diagnostics of thalassemia.
The SNPscan and CNVplex assays utilize variations in PCR product length and fluorescence to identify multiple mutations. In the SNPscan method, we designed three probes per locus: two 5' and one 3', and incorporated allele identification link sequences into one of the 5' probes to distinguish the alleles. The detection system was designed for 67 previously reported loci in the Chinese population for a specific genetic condition. CNVplex identifies deletion types by analyzing the specific positions of probes within the globin gene. This innovative approach enables the detection of six distinct deletional mutations, enhancing the precision of thalassemia diagnostics. We evaluated and refined the methodologies in a training cohort of 100 individuals with confirmed HBA and HBB genotypes. The validation cohort, consisting of 1647 thalassemia patients and 100 healthy controls, underwent a double-blind study. Traditional diagnostic techniques served as the control methods.
In the training set of 100 samples, 10 mutations (Hb QS, Hb CS, Hb Westmead, CD17, CD26, CD41-42, IVS-II-654, --, -α and -α) were identified, consistent with those identified by traditional methods. The validation study showed that SNPscan/CNVplex offered superior molecular diagnostic capabilities for thalassemia, with 100% accuracy compared to 99.43% for traditional methods. Notably, the assay identified three previously undetected mutations in 10 cases, including two deletion mutations (Chinese γ(γδβ) del and SEA-HPFH), and one non-deletion mutation (Hb Q-Thailand).
The SNPscan/CNVplex assay is a cost-effective and user-friendly tool for diagnosing thalassemia, demonstrating high accuracy and reliability, and showing great potential as a primary diagnostic method in clinical practice.
地中海贫血是一种具有严重后果的孟德尔遗传血液疾病，包括残疾和死亡，因此是一个重大的公共卫生关注点。因此，迫切需要精确的诊断技术。我们介绍了两种用于地中海贫血的创新诊断技术，SNPscan 和 CNVplex，旨在增强地中海贫血的分子诊断。
SNPscan 和 CNVplex 检测利用 PCR 产物长度和荧光的变化来识别多种突变。在 SNPscan 方法中，我们在每个基因座设计了三个探针：两个 5'探针和一个 3'探针，并将等位基因识别链接序列整合到其中一个 5'探针中，以区分等位基因。检测系统是为中国人群中 67 个先前报道的基因座设计的，用于特定的遗传条件。CNVplex 通过分析珠蛋白基因内探针的特定位置来识别缺失类型。这种创新方法能够检测到六种不同的缺失突变，提高了地中海贫血诊断的精度。我们在 100 名经证实的 HBA 和 HBB 基因型个体的培训队列中评估和完善了这些方法。验证队列由 1647 名地中海贫血患者和 100 名健康对照组成，进行了双盲研究。传统的诊断技术作为对照方法。
在 100 个样本的培训集中，鉴定出 10 种突变（Hb QS、Hb CS、Hb Westmead、CD17、CD26、CD41-42、IVS-II-654、--、-α和-α），与传统方法鉴定的一致。验证研究表明，SNPscan/CNVplex 为地中海贫血提供了卓越的分子诊断能力，与传统方法的 99.43%相比，准确率达到 100%。值得注意的是，该检测方法在 10 例中发现了三种以前未检测到的突变，包括两种缺失突变（中国γ（γδβ）del 和 SEA-HPFH）和一种非缺失突变（Hb Q-Thailand）。
SNPscan/CNVplex 检测是一种具有成本效益和用户友好的地中海贫血诊断工具，具有很高的准确性和可靠性，在临床实践中具有成为主要诊断方法的巨大潜力。
