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Identification of RFLP G6PD mutations by using microcapillary electrophoretic chips (Experion).
- Source :
-
Journal of separation science [J Sep Sci] 2008 Aug; Vol. 31 (14), pp. 2694-700. - Publication Year :
- 2008
-
Abstract
- Glucose-6-phosphate dehydrogenase (G6PD) deficiency is one of the most well-known human genetic defects, identified in more than 400 million individuals in the world. To date, no commercial kits are available for the mutation screening of this disease. Seventy G6PD-deficient Italian individuals admitted to the Laboratory of Clinical Molecular Biology of Hospital "Agostino Gemelli" of Rome were screened for the most frequent Italian mutations, by means of allele-specific PCR, followed by restriction fragment length electrophoresis. The present study compares two techniques for the identification of restriction patterns: agarose gel electrophoresis versus Experion system. When the first screening was negative, the entire G6PD gene was sequenced using the ABI 3100 Avant Instrumentation. The G6PD variants identified and their frequencies were the following: G6PD Mediterranean (75.7%), G6PD Seattle (7.1%), G6PD A(-) 202 + 376 (7.1%), and G6PD Cassano (2.8%). In addition, we identified by direct sequencing two new mutations, namely Buenos Aires and Rignano. With the Experion method, the size band determination was more accurate than that obtained by gel electrophoresis. The Experion system resulted as a valid, easy, and reproducible diagnostic method for the screening of G6PD mutation as compared with the agarose electrophoretic analysis.
- Subjects :
- DNA Mutational Analysis instrumentation
DNA Mutational Analysis methods
Electrophoresis, Agar Gel instrumentation
Electrophoresis, Agar Gel methods
Electrophoresis, Microchip instrumentation
Female
Gene Frequency
Genetic Testing methods
Humans
Male
Polymerase Chain Reaction instrumentation
Polymerase Chain Reaction methods
Polymorphism, Restriction Fragment Length
Reproducibility of Results
Sensitivity and Specificity
Time Factors
Electrophoresis, Microchip methods
Glucosephosphate Dehydrogenase analysis
Glucosephosphate Dehydrogenase genetics
Mutation
Subjects
Details
- Language :
- English
- ISSN :
- 1615-9314
- Volume :
- 31
- Issue :
- 14
- Database :
- MEDLINE
- Journal :
- Journal of separation science
- Publication Type :
- Academic Journal
- Accession number :
- 18693312
- Full Text :
- https://doi.org/10.1002/jssc.200800216