High resolution melting curve analysis enables rapid and reliable detection of G6PD variants in heterozygous females.
BACKGROUND
Like glucose-6-phosphate dehydrogenase (G6PD) deficient hemizygous males and homozygous females, heterozygous females could also manifest hemolytic crisis, neonatal hyperbilirubinemia or kernicterus upon exposure to oxidative stress induced by certain foods such as fava beans, drugs or infections. Although hemizygous males and homozygous females are easily detected by conventional G6PD enzyme assay method, the heterozygous state could be missed by the conventional methods as the mosaic population of both normal and deficient RBCs circulates in the blood. Thus the present study aimed to apply high resolution melting (HRM) curve analysis approach to see whether HRM could be used as a supplemental approach to increase the chance of detection of G6PD heterozygosity.


RESULTS
Sixty-three clinically suspected females were evaluated for G6PD status using both enzyme assay and HRM analysis. Four out of sixty-three participants came out as G6PD deficient by the enzyme assay method, whereas HRM approach could identify nine participants with G6PD variants, one homozygous and eight heterozygous. Although only three out of eight heterozygous samples had G6PD enzyme deficiency, the HRM-based heterozygous G6PD variants detection for the rest of the samples with normal G6PD enzyme activities could have significance because their newborns might fall victim to serious consequences under certain oxidative stress.


CONCLUSIONS
In addition to the G6PD enzyme assay, HRM curve analysis could be useful as a supplemental approach for detection of G6PD heterozygosity.
['Islam|Md Tarikul|MT|', 'Sarker|Suprovath Kumar|SK|', 'Talukder|Shezote|S|', 'Bhuyan|Golam Sarower|GS|', 'Rahat|Asifuzzaman|A|', 'Islam|Nafisa Nawal|NN|', 'Mahmud|Hasan|H|', 'Hossain|Mohammad Amir|MA|', 'Muraduzzaman|A K M|AKM|', 'Rahman|Jakia|J|', 'Qadri|Syeda Kashfi|SK|', 'Shahidullah|Mohammod|M|', 'Mannan|Mohammad Abdul|MA|', 'Tahura|Sarabon|S|', 'Hussain|Manzoor|M|', 'Saha|Narayan|N|', 'Akhter|Shahida|S|', 'Nahar|Nazmun|N|', 'Begum|Firoza|F|', 'Shirin|Tahmina|T|', 'Akhteruzzaman|Sharif|S|', 'Qadri|Syed Saleheen|SS|', 'Qadri|Firdausi|F|', 'Mannoor|Kaiissar|K|0000-0002-2394-0447']
D000293:Adolescent D002648:Child D002675:Child, Preschool D004252:DNA Mutational Analysis D005260:Female D005954:Glucosephosphate Dehydrogenase D005955:Glucosephosphate Dehydrogenase Deficiency D006579:Heterozygote D006801:Humans D007223:Infant D007231:Infant, Newborn D009154:Mutation D009691:Nucleic Acid Denaturation
High resolution melting curve analysis G6PD heterozygosity Heterozygous G6PD variants Glucose-6-phosphate dehydrogenase deficiency