Application of PCR-DGGE technique in G-6-PD deficiency / 中国当代儿科杂志
Chinese Journal of Contemporary Pediatrics
; (12): 529-532, 2007.
Article
en Zh
| WPRIM
| ID: wpr-325680
Biblioteca responsable:
WPRO
ABSTRACT
<p><b>OBJECTIVE</b>To detect gene mutations of children with glucose-6-phosphorate dehydrogenase (G-6-PD) deficiency and of carriers of G-6-PD deficiency gene with the technique of polymerase chain reaction and denatured gradient gel electrophoresis (PCR-DGGE), and to explore the value of the technique in the diagnosis of G-6-PD deficiency and G-6-PD deficiency gene carrying.</p><p><b>METHODS</b>cDNAs were harvested by reverse transcription method after RNAs had been extracted from peripheral blood of 43 children with G-6-PD deficiency and of their family members (36 lineages). Electrophoresis behaviors of the fragment from exons 11-12 of G-6-PD cDNA were detected with the technique of PCR-DGGE. Gene sequencing was then performed for the abnormal electrophoresis bands.</p><p><b>RESULTS</b>Abnormal electrophoresis bands were found in the 1304-1520 fragment of G-6-PD cDNA in 33 out of 36 family lineages. The G-6-PD/6-PGD ratio was below 1.00 in 9 mothers of patients. Three of them had the G-6-PD/6-PGD ratio lower than 0.50. The PCR-DGGE bands were the same in the 3 mothers. Gene sequencing showed double heterozygote in the 3 mothers, but the maternal carriers of G-6-PD deficiency gene who had normal G-6-PD/6-PGD ratio showed mono-heterozygote in gene sequencing. Three mutational sites were found in the 1304-1520 fragment, i.e., C1311TG1376T and G1388A. The electrophoresis behaviors were different among the 3 gene mutational sites.</p><p><b>CONCLUSIONS</b>PCR-DGGE is a sensitive and reliable technique in the screening of gene mutations. It is useful in the diagnosis of G-6-PD deficiency, especially in the diagnosis of female G-6-PD deficiency gene carrying.</p>
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Índice:
WPRIM
Asunto principal:
Datos de Secuencia Molecular
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Secuencia de Bases
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Reacción en Cadena de la Polimerasa
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Análisis de Secuencia de ADN
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Diagnóstico
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Electroforesis en Gel de Poliacrilamida
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Genética
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Deficiencia de Glucosafosfato Deshidrogenasa
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Métodos
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Mutación
Tipo de estudio:
Diagnostic_studies
Límite:
Female
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Humans
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Male
Idioma:
Zh
Revista:
Chinese Journal of Contemporary Pediatrics
Año:
2007
Tipo del documento:
Article