Accuracy of Reverse Dot-Blot PCR in Detection of Different β-Globin Gene Mutations

Abstract

Prevention programs for b-thalassemia based on molecular diagnosis of heterozygous carriers and/or patients require the use of reliable mutation screening methods. The aim of this study was to compare between direct DNA sequencing, and reverse dot-blot PCR in detection of different b-globin gene mutations in Egyptian children with b-thalassemia. Forty children with b-thalassemia were subjected to mutation analysis, performed by both direct DNA sequencing and b-globin , and cd39 C[T (2.5 %)'', While the genotypes of three patients (6 alleles 7.5 %) were not detected by reverse dot-blot PCR. Mutant alleles detected by direct DNA sequencing were the same as reverse dot-blot PCR method except it revealed the genotypes of 3 undetected patients (one patient was homozygous IVSI-110 G[A, and two patients were homozygous IVS I-1 G[A. Sensitivity of the reverse dot-blot PCR was 92.5 % when compared to direct DNA sequencing for detecting b-thalassemia mutations. Our results therefore suggest that, direct DNA sequencing may be preferred over reverse dot-blot PCR in critical diagnostic situations like genetic counseling for prenatal diagnosis.

Detection of Common Beta Thalassemia Mutations Among Egyptian Patients

“…1). These results were in partial agreement with the findings of El-Fadaly et al (2015), Elmezayen et al (2015) and El-Beshlawy et al (2012).…”

Molecular Basis of Beta Thalassemia Mutations in Egyptian Patients

“…(19). Combined effects of high carrier rates and high frequency of consanguineous marriages make prevalence of β-thalassemia particularly high in Egypt (20).Up to date in Egypt, carrier detection and genetic counseling and carrier identification is essentially presented to families with an affected individual. In these families, the phenotypic and genotypic evaluation is usually performed in these families to determine the prognosis and to offer the comprehensive genetic counseling.…”

Development of a PCR-based dot blot assay for the detection of fowl adenovirus

“…The dot blot assay is a method similar to southern blot that can identify target DNA fragments with high sensitivity ( Cheng et al, 2019 ). Some studies have combined dot blot with PCR to achieve a significant increase in sensitivity ( El-Fadaly et al, 2016 ; Jung and Chae, 2005 ). Therefore, this research also hopes to establish a highly sensitive group Ⅰ FAdV detection method through this method.…”