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Molecular characterization of Chinese G6PD deficiency by using polymerase chain reaction/single strand conformation
B H Chen1, S R Lin, C H Chiang
1Department of Laboratory Medicine, Kaohsiung Medical College Hospital, Taiwan, Republic of China.
Insights
Molecular characterization of glucose-6-phosphate dehydrogenase (G6PD) deficiency in Chinese infants identified seven variants using PCR/SSCP. This method is suitable for screening G6PD deficiency in diverse populations.
Area of Science:
- Genetics
- Biochemistry
- Molecular Biology
Background:
- Glucose-6-phosphate dehydrogenase (G6PD) deficiency is a common genetic disorder.
- Molecular characterization is crucial for understanding G6PD variants and their prevalence.
Purpose of the Study:
- To molecularly characterize G6PD deficient Chinese newborn babies and infants in southern Taiwan.
- To evaluate the suitability of PCR/SSCP for G6PD deficiency screening.
Main Methods:
- Polymerase chain reaction/single strand conformation polymorphism (PCR/SSCP) was employed for molecular screening.
- Direct sequencing was used to confirm mutations identified by PCR/SSCP.
- Analysis included 97 infants (86 male, 11 female) from southern Taiwan.
Main Results:
- Movement shifts were observed in exons 2, 5, 6, and 12, with no shift in exon 9.
- Molecular defects were identified in 90 out of 97 samples, revealing seven variants.
- The most common mutations were at nucleotide 1376 (G-->T, 42.3%) and nucleotide 1388 (G-->A, 34.0%).
- Two novel variants were identified at nucleotide 371 (A-->G) and nucleotide 519 (C-->G).
- No significant difference in G6PD activity was found among the different mutations.
Conclusions:
- PCR/SSCP is a simple, fast, and effective method for molecular screening of G6PD deficiency.
- The study identified common and novel G6PD variants in the Chinese population of Taiwan.
- Findings contribute to understanding the genetic epidemiology of G6PD deficiency.
Abstract:
Using the polymerase chain reaction/single strand conformation polymorphism (PCR/SSCP), we studied the molecular characterization of 97 (86 male and 11 female) glucose-6-phosphate dehydrogenase (G6PD) deficient Chinese newborn babies and infants in southern Taiwan. Movement shifts were clearly seen in exon 2, 5, 6 and 12, respectively. No movement shift was seen in exon 9. Mutation confirmations were followed by direct sequencing. Using this approach we identified the molecular defect in 90 of the 97 above samples. Our results show a total of seven variants, the most common being at nucleotide (nt) 1376 mutation G-->T 42.3% (41/97) and at nt 1388 mutation G-->A 34.0% (33/97). The other mutation sites were at nucleotide 95 A--G (5.2%), nucleotide 392 G-->T (4.1%), nucleotide 493 A-->G (3.1%) and two new variants with mutation at nucleotide 371 A-->G (2.1%) and nucleotide 519 C-->G (2.1%). Still 7.2% (7/97) remained unidentified. There was no significant difference in G6PD activity among the different mutations. Simple and fast, PCR/SSCP may be suitable for molecular screening for G6PD deficiency in Chinese and other people.