- All Journals
- Al-Qadisiyah University
- Basrah University
- Kirkuk University
- Mosul University
- The Iraqi Borad for Medical Specialization
- University of Anbar
- University of Technology
- Al Mustafa University
- Al-Zahraa University for Women
- Alnoor University
- Kunooz University
- Misan University
- Ninevah University
- Tikrit University
- University of Al-Hamdaniya
- University of Kerbala
Molecular Characterization of Severe G6PD Deficiency in Hyperbilirubinemic Neonates in Karbala : Iraq…… | ||
| Kerbala Journal of Medicine | ||
| Article 15, Volume 2, Issue 5, 2009, Pages 663-680 PDF (0 K) | ||
| Authors | ||
| Fadhil J. Al-Touma; William M. Frankool | ||
| Abstract | ||
| Objective: The objective of this study was to investigate the molecular basis of glucose-6-phosphate dehydrogenase (G6PD) genes in hyperbilirubinemic neonates in Karbala province of Iraq by using molecular methods (genomic DNA extraction, PCR and RFLP analysis) and then to investigate the type of G6PD variant predominantly present. Methods: The study included a total of 253 full-term male neonates, 197 of them associated with severe hyperbilirubinemia which were admitted in Karbala Teaching Hospital of Pediatrics during the period from 1st October 2007 to 14th July 2008 with age ranged between 1 – 28 days, their TSB levels ≥ 15 mg/dl, and another 53 neonates were used as control group. The blood sample taken from each neonate was divided into two aliquots: the first aliquot was used for total and conjugated serum bilirubin (TSB and SCB), G6PD activity. The second aliquot was used for molecular analysis including genomic DNA extraction and then application of polymerase chain reaction (PCR) and restriction fragment length polymorphism (RFLP) protocols. Results and Discussion: Severe hyperbilirubinemic neonates were screened for erythrocyte G6PD enzyme activity measurements, severe G6PD deficiency was detected in 18 of the total 197 hyperbilirubinemic neonates included and their activity levels was significantly decreased (P < 0.05) to 0.56 ± 0.32 U/g Hb. The incidence of severe G6PD deficiency identified was found 9.14%. TSB levels were markedly elevated to (20.26 ± 4.96 mg/dl), whereas the mean ± SD values of SCB were significantly lower than that found in controls (P < 0.05) and reached to 0.053 ± 0.046 mg/dl , and it was undetectable in 5 of 18 neonates (27.78%) with severe G6PD deficiency which imply a partial defect of bilirubin conjugation. The molecular part of the study involved the extraction of genomic DNA from hyperbilirubinemic neonates with severe G6PD deficiency which detected by agarose electrophoresis and then amplified by PCR and finally was subjected to digestion by endonuclease restriction enzymes to create RFLP to enable the detection of mutation that caused G6PD deficiency. The overall majority of affected severe G6PD neonates with hyperbilirubinemia in Kerbala province : Iraq were due to G6PD Med variant (C563T, Ser 188 Phe) in which 17 out of 18 (94.4%) have this type of mutation, and only one G6PD A- variant (5.56%) (G202A ; A376G mutations) was diagnosed. Conclusion: The predominant G6PD gene detected in hyperbilirubinemic neonate with severe G6PD deficiency in Kerbala province was G6PD Med. | ||
| Keywords | ||
| Hyperbilirubinemia; PD gene; Polymerase Chain Reaction; RFLP | ||
|
Statistics Article View: 42 PDF Download: 52 |
||