Rapid screening for glucose-6-phosphate dehydrogenase deficiency and haemoglobin polymorphisms in Africa by a simple high-throughput SSOP-ELISA method.
Rapid screening for glucose-6-phosphate dehydrogenase deficiency and haemoglobin polymorphisms in Africa by a simple high-throughput SSOP-ELISA method.
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DOI：
10.1186/1475-2875-4-61
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发表时间：
2005-12-15
期刊：
影响因子：
3
通讯作者：
Alifrangis M
中科院分区：
文献类型：
--
作者：
Enevold A;Vestergaard LS;Lusingu J;Drakeley CJ;Lemnge MM;Theander TG;Bygbjerg IC;Alifrangis M
血红蛋白β-珠蛋白（HbB）和葡萄糖-6-磷酸脱氢酶（G6 PD）基因的突变导致广泛的人类遗传疾病，如镰状细胞病和G6 PD缺乏症。在撒哈拉以南非洲，每个基因的一些主要多态性变体导致了这些缺陷的大部分。如果能够以有限的成本获得准确的筛查方法，就更容易在更大范围内检查这些独立的遗传性疾病的临床重要性、它们与疟疾发病机制和先天耐药性的可能联系以及它们与抗疟药物治疗的相关性。建立了一种简便、快速的检测HbB和G6 PD基因中最显著的单核苷酸多态性（SNPs）的方法。该方法能够检测不同的血红蛋白多态性A、S、C和E，以及G6 PD多态性B、A和A-基于PCR扩增，然后使用对SNP变体特异的序列特异性寡核苷酸探针（SSOP）进行杂交步骤，并通过ELISA定量。SSOP-ELISA方法被认为是特异性的，并与常用的PCR-RFLP技术进行了比较。在坦桑尼亚一个高传播地区的90个现场样本中，98%（血红蛋白）和95%（G6 PD）获得了相同的结果，用于验证新技术。新方法的简单性和准确性使其适合在资源有限的情况下应用。通过监测与疾病流行病学相关的基因型频率，它将成为研究目的的一个有价值的工具。
Mutations in the haemoglobin beta-globin (HbB) and glucose-6-phosphate dehydrogenase (G6PD) genes cause widespread human genetic disorders such as sickle cell diseases and G6PD deficiency. In sub-Saharan Africa, a few predominant polymorphic variants of each gene account for a majority of these deficiencies. Examining at a larger scale the clinical importance of these independent genetic disorders, their possible association with malaria pathogenesis and innate resistance, and their relevance for antimalarial drug treatment, would be easier if an accurate screening method with limited costs was available. A simple and rapid technique was developed to detect the most prominent single nucleotide polymorphisms (SNPs) in the HbB and G6PD genes. The method is able to detect the different haemoglobin polymorphisms A, S, C and E, as well as G6PD polymorphisms B, A and A- based on PCR-amplification followed by a hybridization step using sequence-specific oligonucleotide probes (SSOPs) specific for the SNP variants and quantified by ELISA. The SSOP-ELISA method was found to be specific, and compared well to the commonly used PCR-RFLP technique. Identical results were obtained in 98% (haemoglobin) and 95% (G6PD) of the tested 90 field samples from a high-transmission area in Tanzania, which were used to validate the new technique. The simplicity and accuracy of the new methodology makes it suitable for application in settings where resources are limited. It would serve as a valuable tool for research purposes by monitoring genotype frequencies in relation to disease epidemiology.
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影响因子：
64.8
作者：
Modiano, D;Luoni, G;Coluzzi, M
通讯作者：
Coluzzi, M
DOI：
10.4269/ajtmh.2005.72.155
发表时间：
2005-02-01
影响因子：
3.3
作者：
Alifrangis, M;Enosse, S;Bygjerg, IBC
通讯作者：
Bygjerg, IBC
DOI：
10.4269/ajtmh.2003.68.186
发表时间：
2003-02-01
影响因子：
3.3
作者：
Mombo, LE;Ntoumi, F;Krishnamoorthy, R
通讯作者：
Krishnamoorthy, R
DOI：
10.1016/0035-9203(54)90101-7
发表时间：
1954-01-01
影响因子：
2.2
作者：
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通讯作者：
ALLISON, AC
影响因子：
1.8
作者：
Bouanga, JC;Mouélé, R;Galactéros, F
通讯作者：
Galactéros, F