A mismatched-primer polymerase chain reaction-restriction fragment length polymorphism strategy for rapid screening of the polyadenylation signal mutation alpha(T-Saudi) (AATAAA-->AATAAG) in the alpha2-globin gene.

The most common nondeletional alpha-thalassemia allele, namely alpha(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 beta-thalassemia and sickle cell disease, also quite prevalent in these regions. Recognition of the alpha(T-Saudi) allele has so far relied on technically-demanding procedures. Here we report a simple, rapid, and robust polymerase 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.

[1]  Y. Fei,et al.  Hb H disease caused by a homozygosity for the AATAAA-->AATAAG mutation in the polyadenylation site of the alpha 2-globin gene: hematological observations. , 1992, Acta haematologica.

[2]  D. Higgs,et al.  A PCR‐based strategy to detect the common severe determinants of α thalassaemia , 1992 .

[3]  S. Goodbourn,et al.  α-Thalassaemia caused by a polyadenylation signal mutation , 1983, Nature.

[4]  E. Baysal,et al.  Detection of common deletional α‐thalassemia‐2 determinants by PCR , 1994 .

[5]  J. Clegg,et al.  The polyadenylation site mutation in the alpha-globin gene cluster. , 1988, Blood.

[6]  S. Humphries,et al.  PCR induction of a TaqI restriction site at any CpG dinucleotide using two mismatched primers (CpG-PCR). , 1996, Genome research.

[7]  C. Dodé,et al.  Locus assignment of human a globin mutations by selective amplification and direct sequencing , 1990, British journal of haematology.

[8]  M. Takeda,et al.  Mutational screening of APP gene in patients with early‐onset Alzheimer disease utilizing mismatched PCR‐RFLP , 1996, Clinical genetics.

[9]  M. Tzetis,et al.  Characterization of nondeletion α‐thalassemia mutations in the Greek population , 1993 .

[10]  A. Adekile,et al.  Morbidity, βs Haplotype and α-Globin Gene Patterns among Sickle Cell Anemia Patients in Kuwait , 1996 .

[11]  P. Fortina,et al.  Human α‐Thalassemia syndromes: Detection of molecular defects , 1996 .

[12]  M. Losekoot,et al.  A novel polyadenylation signal mutation in the α2‐globin gene causing α thalassaemia , 1994 .