Mirasena Saisiri, Shimbhu Dawan, Sanguansermsri Mondhon, Sanguansermsri Torpong
Department of Biochemistry, Faculty of Medical Science, Naresuan University, Phitsanulok, Thailand.
Hemoglobin. 2008;32(4):403-9. doi: 10.1080/03630260701798391.
We developed two sets of a multiplex amplification refractory mutation system (M-ARMS) assay to identify specific beta-thalassemia (beta-thal) mutations that are common in Thailand. The first one was for the detection of mutants with codon 17 (A>T), IV S-I-1 (G >T)), codons 41/42 (-TCT T) and codons 71/72 (+A), while the second one was for the -87 (C>A), -28 (A>G) and IVS-II-654 (C>T). Application of the proposed assay to 282 persons with beta-thal trait revealed a positive result in 276 cases (97.8%). There were 258 cases (91.5%) positive for the set 1 M-ARMS assay and 18 cases (6.4%) were positive for set 2. Six cases (2.2%) were negative for both sets 1 and 2, and were further characterized by DNA sequencing. The mutations detected by the set 1 M-ARMS assay were 113 cases (40.1%) of codons 41/42, 95 (33.7%) of codon 17, 41 (14.5%) of IVS-I-1 and nine cases (3.2%) of codons 71/72, while by set 2 there were 12 cases (4.2%) of -28, four cases(1.4%) of -87 and two cases (0.7%) of IVS-II-654. Mutations undetectable by M-ARMS assay were two cases of codons 27/28 (+C), one case of codon 35 (C>A), one of codon 43 (G>T), one of -31 (A>G) and one of IVS-I-5 (C>G). The M-ARMS assay proved to be a valuable tool for the analysis of beta-thal mutations. The method is robust, accurate, simple, speedy and cost-effective. The application of this assay will facilitate genetic counseling and prenatal diagnosis for severe thalassemia in high-risk pregnancies.
我们开发了两套多重扩增不应性突变系统（M-ARMS）检测方法，以鉴定泰国常见的特定β地中海贫血（β-地贫）突变。第一套用于检测密码子17（A>T）、IVS-I-1（G>T）、密码子41/42（-TCT T）和密码子71/72（+A）的突变体，而第二套用于检测-87（C>A）、-28（A>G）和IVS-II-654（C>T）。将所提出的检测方法应用于282例β-地贫特征患者，276例（97.8%）检测结果呈阳性。第一套M-ARMS检测方法阳性的有258例（91.5%），第二套阳性的有18例（6.4%）。两套检测方法均为阴性的有6例（2.2%），并通过DNA测序进一步鉴定。第一套M-ARMS检测方法检测到的突变中，密码子41/42的有113例（40.1%），密码子17的有95例（33.7%），IVS-I-1的有41例（14.5%），密码子71/72的有9例（3.2%）；而第二套检测到的突变中，-28的有12例（4.2%），-87的有4例（1.4%），IVS-II-654的有2例（0.7%）。M-ARMS检测方法未检测到的突变有密码子27/28（+C）的2例、密码子35（C>A）的1例、密码子43（G>T）的1例、-31（A>G）的1例和IVS-I-5（C>G）的1例。M-ARMS检测方法被证明是分析β-地贫突变的一种有价值的工具。该方法稳健、准确、简单、快速且具有成本效益。该检测方法的应用将有助于高危妊娠中重型地中海贫血的遗传咨询和产前诊断。
