Mismatched-Pruner Polymerase Chain Reaction-Restriction Fragment Length Polymorphism Strategy for Rapid Screening of the Polyadenylation Signal Mutation ?^T-Saudi (AATAAA?AATAAG) in the ?2-Globin Gene

N. Jassian1,2, S. Al-Arrayed1, N. Gerard 2, H. Al-Mukiaarraq1,A Al-Ajami1, R. Ramasawmy2, and

R. Krishnatnoorthy2 Salmaniya Medical Center, Manama, Bahrain 2JNSERM U 458, H?pital Robert Debr? 75019 Paris,France

ABSTRACT

The most common nondeietional ?-thalassemia allele, namely ?^T?Saudi (AATAAA?AATAAG) in the Arabian peninsula and neighboring countries is responsible for a number of cases of Hb H disease. It is expected to alter significantly the clinical manifestations of ?-thalassemia and sickle cell disease, also qulle prevalent in these regions. Recognition of the ? ^{T ? Saudi} allele has so far relied on technically-demanding procedures. here we report a simple, rapid, and robust polynierase chain reaction-based detection procedure for this allele. This Involves priming of the polymerase chain reaction with a deliberately introduced mismatch in one of the primers so that the mutant allele, after amplification, would Introduce a StuI restriction enzyme site, the presence of which can be recognized by digesting the polymerase chain reaction product with. This enzyme

Correspondence should be addressed to Dr. Rajagopal Krishnamoorth, INSERM U 458, HOPITAL Report Debre, 48 Boulevard Serurier, 75019 Paris, France; TEL: +33(01) 40031901; FAX: +33(01) 40031903; e-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it