Resumen
La anemia falciforme es una hemoglobinopatía hereditaria caracterizada por la presencia de hemoglobina S (HbS), que se estima afecta aproximadamente al 0.04% de la población panameña. Su severidad clínica está modulada por los haplotipos del gen β-globina —Benín, Bantú, Senegal, Camerún y Árabe-hindú—, cuya caracterización no es posible mediante la cromatografía líquida de alta resolución (HPLC) utilizada en el Programa de Tamizaje Neonatal del Hospital del Niño Dr. José Renán Esquivel.
El objetivo de este estudio fue estandarizar la técnica de PCR punto final para la identificación de dichos haplotipos en 22 pacientes con HbSS confirmado, correspondientes al periodo enero 2020-agosto 2025. Se extrajo ADN genómico utilizando el kit PureLink (Invitrogen) y se amplificaron seis regiones específicas del gen β-globina. Los productos fueron digeridos con las enzimas DdeI, HindIII, HincII, HinfI, y visualizados en geles de agarosa al 2%.
Se identificaron tres haplotipos: Senegal (45.5%), Bantú (40.9%) y Benín (13.6%); los haplotipos Camerún y árabe-hindú no fueron detectados. El protocolo estandarizado demostró ser reproducible y confiable, constituyendo una herramienta molecular complementaria para la subclasificación de pacientes con HbSS.
Palabras clave: Anemia Falciforme, haplotipos, PCR punto final, tamizaje neonatal.
Abstract
Sickle cell anemia is an inherited hemoglobinopathy characterized by the presence of hemoglobin S (HbS), which is estimated to affect approximately 0.04% of the Panamanian population. Its clinical severity is modulated by the haplotypes of the β-globin gene—Benin, Bantu, Senegal, Cameroon, and Arab-Hindu—whose characterization is not possible using high-performance liquid chromatography (HPLC) as employed in the Neonatal Screening Program at Dr. José Renán Equivel Children’s Hospital.
The objective of this study was to standardize the endpoint PCR technique for the identification of these haplotypes in 22 patients with confirmed HbSS, corresponding to the period January 2020–August 2025. Genomic DNA was extracted using the PureLink kit (Invitrogen), and six specific regions of the β-globin gene were amplified. The products were digested with the enzymes DdeI, HindIII, HincII, and HinfI, and visualized on 2% agarose gels.
Three haplotypes were identified: Senegal (45.5%), Bantu (40.9%), and Benin (13.6%); the Cameroon and Arab-Hindu haplotypes were not detected. The standardized protocol proved to be reproducible and reliable, constituting a complementary molecular tool for the subclassification of patients with HbSS.
Keywords: sickle cell disease, β-globin haplotypes, endpoint PCR, neonatal screening.