• Emuejevoke T Toye, Guido van Marle, Wendy Hutchins, Olayinka Abgabiaje, and Joy Okpuzor.
• University of Lagos, Department of Cell Biology and Genetics.
• Afr Health Sci. 2018 Dec 1; 18 (4): 995-1002.

BackgroundSickle cell anaemia (SCA) is a hereditary blood disorder caused by a single mutation in the haemoglobin gene. The disease burden of SCA is highest in Nigeria. The allele specific polymerase chain reaction (ASPCR) method is applicable for the direct detection of known single nucleotide polymorphisms (SNPs).ObjectiveTo investigate the use of the single tube ASPCR as an accurate and affordable method for SCA screening in Nigeria.MethodsDNA was extracted from study subjects with normal haemoglobin, HbAA (20), sickle cell anaemia, HbSS (20) and carriers, HbAS (1). Haemoglobin was genotyped by ASPCR using two primer sets that amplifies the wildtype and mutant haemoglobins in each sample. Amplicon sizes were analyzed by gel electrophoresis.ResultsAmplicons were visible after electrophoresis at regions 517 base pair (bp) for HbA and 267 bp for HbS. ASPCR correctly and unambiguously detected the presence or absence of haemoglobins A and S from all samples collected, demonstrating its accuracy and precision for the screening of SCA.ConclusionThis study validates ASPCR as an effective, low cost approach for the clinical screening of SCA in Nigeria. ASPCR is also applicable for other genetic diseases, paternity testing, and forensics where more expensive fluorescence-based approaches are not obtainable.

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