Department of Pathology, Maha Chakri Sirindhorn Medical Center, Faculty of Medicine, Srinakharinwirot University, Ongkharak, Nakhon Nayok, Thailand.
Centre for Research and Development of Medical Diagnostic Laboratories, Faculty of Associated Medical Sciences, Khon Kaen University, Khon Kaen, Thailand.
PLoS One. 2022 Apr 28;17(4):e0267832. doi: 10.1371/journal.pone.0267832. eCollection 2022.
Identification of α0-thalassemia (SEA and THAI deletions) is essential in preventing and controlling of severe thalassemia diseases. We have developed the LAMP colorimetric assays for the detection of these two thalassemia defects and validated them in population screening and prenatal diagnosis.
Three LAMP colorimetric assays specific for α0-thalassemia (SEA deletion), α0-thalassemia (THAI deletion) and normal DNA sequence were developed. These assays were validated on 341 subjects who had initial thalassemia screening positive and various thalassemia genotypes. Prenatal diagnosis of α0-thalassemia (SEA deletion) was done on 33 fetuses at risk of having Hb Bart's hydrops fetalis syndrome.
The LAMP colorimetric assays for α0-thalassemia (SEA and THAI deletions) could be clearly interpreted by naked eyes. The assay for α0-thalassemia (SEA deletion) showed a 100% (62/62 x 100) sensitivity and 98.2% (274/279 x 100) specificity whereas, that of the α0-thalassemia (THAI deletion) showed 100% (1/1 x 100) sensitivity and 99.7% (339/340 x 100) specificity. We obtained a 100% concordant prenatal diagnosis results using LAMP assays of α0-thalassemia (SEA deletion) in 33 fetuses as compared to the conventional PCR analysis.
The LAMP colorimetric assays developed are simple, rapid, and do not require sophisticated equipment. Inclusion of the LAMP tests in the existing screening protocol significantly reduce the screening cost and the molecular analysis workload, which should prove useful in the prevention and control program of hemoglobinopathies in the region.
鉴定α0-地中海贫血（SEA 和 THAI 缺失）对于预防和控制严重地中海贫血疾病至关重要。我们已经开发了用于检测这两种地中海贫血缺陷的 LAMP 比色测定法，并在人群筛查和产前诊断中进行了验证。
开发了三种针对α0-地中海贫血（SEA 缺失）、α0-地中海贫血（THAI 缺失）和正常 DNA 序列的 LAMP 比色测定法。这些测定法在 341 名最初地中海贫血筛查阳性且具有各种地中海贫血基因型的受试者中进行了验证。对 33 名有 Hb Bart's 水肿胎儿综合征风险的胎儿进行了产前诊断α0-地中海贫血（SEA 缺失）。
α0-地中海贫血（SEA 和 THAI 缺失）的 LAMP 比色测定法可以通过肉眼清晰判断。用于检测α0-地中海贫血（SEA 缺失）的测定法显示出 100%（62/62 x 100）的灵敏度和 98.2%（274/279 x 100）的特异性，而用于检测α0-地中海贫血（THAI 缺失）的测定法则显示出 100%（1/1 x 100）的灵敏度和 99.7%（339/340 x 100）的特异性。与传统的 PCR 分析相比，我们使用 LAMP 测定法对 33 名胎儿进行α0-地中海贫血（SEA 缺失）的产前诊断结果完全一致。
开发的 LAMP 比色测定法简单、快速，且不需要复杂的设备。将 LAMP 测试纳入现有的筛查方案可显著降低筛查成本和分子分析工作量，这对于该地区的血红蛋白病预防和控制计划应该是有用的。
