African Health Sciences
Makerere University Medical School
Vol. 18, No. 4, 2018, pp. 995-1002
Bioline Code: hs18123
Full paper language: English
Document type: Study
Document available free of charge
African Health Sciences, Vol. 18, No. 4, 2018, pp. 995-1002
© Copyright 2018 - Toye et al.
Single tube allele specific PCR: a low cost technique for molecular screening of sickle cell anaemia in Nigeria|
Toye, Emuejevoke T; Marle, Guido van; Hutchins, Wendy; Abgabiaje, Olayinka & Okpuzor, Joy
Background: Sickle 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).
Objective: To investigate the use of the single tube ASPCR as an accurate and affordable method for SCA screening in Nigeria.
Methods: DNA 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.
Results: Amplicons 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.
Conclusion: This 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.
Sickle cell anaemia; polymerase chain reaction; molecular diagnostic techniques