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.
Jassim
N
N
Salmaniya Medical Center, Manama, Bahrain.
Al-Arrayed
S
S
Gerard
N
N
Al-Mukharraq
H
H
Al-Ajami
A
A
Ramasawmy
R
R
Krishnamoorthy
R
R
D000293:Adolescent; D000328:Adult; D000483:Alleles; D020137:Base Pair Mismatch / Q000235:genetics; D002648:Child; D002675:Child, Preschool; D005602:France / Q000453:epidemiology; D005820:Genetic Testing; D005838:Genotype; D005914:Globins / Q000737:chemistry / Q000235:genetics*; D006720:Homozygote; D006801:Humans; D008875:Middle Aged; D008877:Middle East / Q000208:ethnology; D017354:Point Mutation; D011061:Poly A / Q000235:genetics* / Q000378:metabolism; D016133:Polymerase Chain Reaction / Q000379:methods; D012150:Polymorphism, Restriction Fragment Length; D013696:Temperature; D017085:alpha-Thalassemia / Q000175:diagnosis* / Q000235:genetics*
D011061:Poly A; D005914:Globins