Single tube allele specific PCR: a low cost technique for molecular screening of sickle cell anaemia in Nigeria
| Autor: | Emuejevoke T. Toye, Wendy Hutchins, Joy Okpuzor Okpuzor, Guido van Marle, Olayinka Abgabiaje |
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| Rok vydání: | 2019 |
| Předmět: |
Sickle cell anaemia
polymerase chain reaction molecular diagnostic techniques polymerase chain reaction 030231 tropical medicine Cell Hemoglobin Sickle Nigeria Single-nucleotide polymorphism Anemia Sickle Cell Polymorphism Single Nucleotide Sensitivity and Specificity law.invention 03 medical and health sciences 0302 clinical medicine law medicine Humans Mass Screening Gene Polymerase chain reaction Alleles Gel electrophoresis Genetics business.industry Sickle cell anaemia Reproducibility of Results Hemoglobin A General Medicine Articles Amplicon medicine.anatomical_structure 030220 oncology & carcinogenesis molecular diagnostic techniques Primer (molecular biology) Variants of PCR business |
| Zdroj: | African Health Sciences African Health Sciences; Vol 18, No 4 (2018); 995-1002 |
| ISSN: | 1729-0503 1680-6905 |
| Popis: | 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. Keywords: Sickle cell anaemia, polymerase chain reaction, molecular diagnostic techniques. |
| Databáze: | OpenAIRE |
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