Comparison of a semiquantitative and a quantitative method for neonatal screening of glucose 6-phosphate dehydrogenase deficiency with subsequent confirmation by allelic discrimination assays

Abstract

Glucose 6-phosphate dehydrogenase (G6PD) deficiency is a X-linked disorder causing neonatal jaundice and haemolysis on oxidative stress. We compared the currently used fluorescent spot test (FST) against a quantitative assay (OSMMR2000-D) and molecular testing methods for screening and identification of G6PD deficient neonates. A total of 1371 cord blood samples were tested using the semiquantitative FST and quantitative G6PD assay. Positive FST samples were subjected to allelic discrimination assays for G6PD Viangchan, Mediterranean, Canton and Kaiping, which encompass over 75% of the G6PD mutations detected in our population. Positive FST results were seen in 86 (6.3%) of the samples tested. Mean G6PD level using the quantitative assay was 15.3 U/g haemoglobin (Hgb) with 60% and 20% enzyme activity 9.2 and 3.1 U/g Hgb, respectively. Twenty-one of the 86 samples detected as positive by FST had activity above 9.2 U/g Hgb, 10 of which were identified as harbouring one of the four mutations tested. The quantitative assay used in this study, using cord blood samples may cause false negative results if limits are established as proposed. We propose that the FST remains an effective screening tool for G6PD deficiency and can be supplemented by rapid real time molecular identification methods for confirmation.