Kompetitive allele specific PCR (KASP) based genotyping of sickle gene in the selected sub-ethnic tribal population of Gujarat and Madhya Pradesh.

BACKGROUND

Indian tribal population is more vulnerable to sickle cell disease (SCD), an autosomal recessive disease caused by a single A>T transversion mutation in the HBB gene. Current study aims to develop a novel Kompetitive allele specific PCR (KASP) assay for genotyping of sickle gene in selected tribal communities of India.

PATIENTS AND

METHODS

The study was conducted from 2021 to 2023 involving 583 self-declared healthy individuals from four tribal communities, i.e. Korku, Kol, and Sahariya of Madhya Pradesh and Dongri bhil of Gujarat. The sickle cell genotyping was performed using KASP and PCR-RFLP methods and further validated by Sanger sequencing. Various hematological and biochemical parameters were also studied and compared for their association with identified genotypes.

RESULTS

Prevalence of sickle cell trait (SCT) was observed as 4.67%, 9.33%, and 16.41% in Kol, Korku, and Dongri bhil, respectively. The SCD prevalence of 0.7% was observed in Dongri bhil. The KASP assay reflects as a novel, high-throughput, and cost-effective method for effective screening. RBC, folic acid, fetal Hb (HBF), and bilirubin have shown significant association with sickle gene mutation.

CONCLUSIONS

KASP assay offers a rapid, precise, and cost-effective method for screening in tribal populations, which can help in early identification, management, and elimination of disease.