Tan K L, Tan J A, Wong Y C, Wee Y C, Thong M K, Yap S F
Department of Allied Health Sciences, Faculty of Medicine, University of Malaya, Kuala Lumpur, Malaysia.
Genet Test. 2001 Spring;5(1):17-22. doi: 10.1089/109065701750168626.
Beta-thalassemia major patients have chronic anemia and are dependent on blood transfusions to sustain life. Molecular characterization and prenatal diagnosis of beta3-thalassemia is essential in Malaysia because about 4.5% of the population are heterozygous carriers for beta-thalassemia. The high percentage of compound heterozygosity (47.62%) found in beta-thalassemia major patients in the Thalassaemia Registry, University of Malaya Medical Centre (UMMC), Malaysia, also supports a need for rapid, economical, and sensitive protocols for the detection of beta-thalassemia mutations. Molecular characterization of beta-thalassemia mutations in Malaysia is currently carried out using ARMS, which detects a single beta-thalassemia mutation per PCR reaction. We developed and evaluated Combine amplification refractory mutation system (C-ARMS) techniques for efficient molecular detection of two to three beta-thalassemia mutations in a single PCR reaction. Three C-ARMS protocols were evaluated and established for molecular characterization of common beta-thalassemia mutations in the Malay and Chinese ethnic groups in Malaysia. Two C-ARMS protocols (cd 41-42/IVSII #654 and -29/cd 71-72) detected the beta-thalassemia mutations in 74.98% of the Chinese patients studied. The CARMS for cd 41-42/IVSII #654 detected beta-thalassemia mutations in 72% of the Chinese families. C-ARMS for cd 41-42/IVSI #5/cd 17 allowed detection of beta-thalassemia mutations in 36.53% of beta-thalassemia in the Malay patients. C-ARMS for cd 41-42/IVSI #5/cd 17 detected beta-thalassemia in 45.54% of the Chinese patients. We conclude that C-ARMS with the ability to detect two to three mutations in a single reaction provides more rapid and cost-effective protocols for beta-thalassemia prenatal diagnosis and molecular analysis programs in Malaysia.
重型β地中海贫血患者患有慢性贫血，依赖输血维持生命。在马来西亚，β3地中海贫血的分子特征分析和产前诊断至关重要，因为约4.5%的人口是β地中海贫血的杂合子携带者。马来西亚马来亚大学医学中心（UMMC）地中海贫血登记处的重型β地中海贫血患者中发现的高复合杂合率（47.62%）也表明，需要快速、经济且灵敏的方案来检测β地中海贫血突变。马来西亚目前使用扩增阻滞突变系统（ARMS）进行β地中海贫血突变的分子特征分析，该方法每次PCR反应只能检测一种β地中海贫血突变。我们开发并评估了联合扩增阻滞突变系统（C-ARMS）技术，以在单个PCR反应中高效分子检测两到三种β地中海贫血突变。评估并建立了三种C-ARMS方案，用于马来西亚马来族和华族常见β地中海贫血突变的分子特征分析。两种C-ARMS方案（cd 41-42/IVSII #654和-29/cd 71-72）在74.98%的受研究华裔患者中检测到β地中海贫血突变。针对cd 41-42/IVSII #654的C-ARMS在72%的华裔家庭中检测到β地中海贫血突变。针对cd 41-42/IVSI #5/cd 17的C-ARMS在36.53%的马来族β地中海贫血患者中检测到β地中海贫血突变。针对cd 41-42/IVSI #5/cd 17的C-ARMS在45.54%的华裔患者中检测到β地中海贫血。我们得出结论，能够在单个反应中检测两到三种突变的C-ARMS为马来西亚的β地中海贫血产前诊断和分子分析项目提供了更快速且具成本效益的方案。
