AS-PCR IN MOLECULAR DIAGNOSTICS: A SYSTEMATIC REVIEW OF APPLICATIONS IN GENETIC DISEASE SCREENING

Abstract

Allele-Specific Polymerase Chain Reaction (AS-PCR) is a highly specific molecular technique widely employed for the detection of single nucleotide polymorphisms (SNPs) and known point mutations in genetic disease screening. This study presents a comprehensive meta-analysis to evaluate the diagnostic accuracy, clinical applicability, and global integration of AS-PCR across diverse genetic conditions, including thalassemia, cystic fibrosis, sickle cell disease, hereditary cancers, and somatic mutations. Utilizing the PRISMA 2020 framework, peer-reviewed articles published between 2000 and 2024 were systematically identified and analyzed. Eligibility criteria included empirical studies reporting sensitivity, specificity, or comparative diagnostic performance of AS-PCR against standard molecular methods. A random-effects model was used to compute pooled sensitivity, specificity, and diagnostic odds ratios, while heterogeneity and publication bias were assessed through I² statistics and funnel plot analysis. The meta-analysis revealed consistently high diagnostic performance of AS-PCR, with pooled sensitivity and specificity values exceeding 95%, confirming its utility as a frontline diagnostic tool. The findings also highlighted AS-PCR’s versatility in low-resource settings, its enhanced sensitivity in digital and microfluidic formats, and its integration into national screening and reproductive health programs. Overall, this study affirms AS-PCR’s critical role in precision diagnostics and public health genomics, particularly for targeted mutation screening and cost-sensitive healthcare environments.