Tepakhan Wanicha, Jomoui Wittaya
Department of Pathology, Faculty of Medicine, Prince of Songkla University, Songkhla, Thailand.
Department of Pathology, Maha Chakri Sirindhorn Medical Center, Faculty of Medicine, Srinakharinwirot University, Nakhon Nayok, Thailand.
Heliyon. 2021 Nov 12;7(11):e08372. doi: 10.1016/j.heliyon.2021.e08372. eCollection 2021 Nov.
β-thalassemia is an inherited disorder that is reported worldwide. Two common β-thalassemia mutations (3.5 kb and 45 kb deletions) are prevalent in Southeast Asia and Thailand. Identification of these defects is essential to population screening and prenatal diagnosis. We aimed to develop colorimetric LAMP based on a phenol red indicator and validate it on various thalassemia genotypes.
Colorimetric LAMP assays for detecting β-thalassemia 3.5- and 45-kb deletions were developed and validated on 254 routine clinical samples. The results of the assays could be interpreted by the naked eye and compared with the gold standard gap-PCR.
A total of 254 samples related to seven phenotypes and 27 different genotype groups showed 100% concordance between the colorimetric LAMP assays and gap-PCR for detecting β-thalassemia (3.5- and 45-kb deletions). The sensitivity, specificity, NPV, and PPV were calculated as 100% for both β-thalassemia 3.5- and 45-kb deletion detection. The comparison of the usefulness of colorimetric LAMP assays and conventional methods was demonstrated in this study.
The developed colorimetric LAMP assays are rapid, simple, and highly cost effective and can be interpreted by the naked eye. These assays should be applied for screening deletional β-thalassemia in routine settings or small community hospitals in remote areas where thalassemia is highly heterogeneous.
β地中海贫血是一种遗传性疾病，在全球范围内均有报道。两种常见的β地中海贫血突变（3.5 kb和45 kb缺失）在东南亚和泰国较为普遍。识别这些缺陷对于人群筛查和产前诊断至关重要。我们旨在开发基于酚红指示剂的比色环介导等温扩增技术（LAMP），并在各种地中海贫血基因型上进行验证。
开发了用于检测β地中海贫血3.5 kb和45 kb缺失的比色LAMP检测方法，并在254份常规临床样本上进行了验证。检测结果可用肉眼解读，并与金标准缺口PCR进行比较。
总共254份与7种表型和27个不同基因型组相关的样本显示，在检测β地中海贫血（3.5 kb和45 kb缺失）时，比色LAMP检测方法与缺口PCR之间的一致性为100%。β地中海贫血3.5 kb和45 kb缺失检测的灵敏度、特异性、阴性预测值和阳性预测值均计算为100%。本研究对比了比色LAMP检测方法和传统方法的实用性。
所开发的比色LAMP检测方法快速、简单且成本效益高，可用肉眼解读。这些检测方法应应用于常规环境或地中海贫血高度异质性的偏远地区的小型社区医院，以筛查缺失型β地中海贫血。
