Detection and haplotype differentiation of Southeast Asian alpha-thalassemia using polymerase chain reaction and a piezoelectric biosensor immobilized with a single oligonucleotide probe.
DNA-based diagnosis of alpha-thalassemias routinely relies on polymerase chain reaction (PCR) and gel electrophoresis. Here, we developed a new procedure for the detection and haplotype differentiation of Southeast Asian (SEA) alpha-thalassemia using a 3-primer system for PCR coupling with a DNA-based piezoelectric biosensor. PCR products amplified from genomic DNA were differentiated directly by using a quartz crystal microbalance immobilized with a single oligonucleotide probe. The frequency changes after hybridization of the PCR products amplified from a representative sample of normal alpha-globin, SEA alpha-thalassemia heterozygote, and homozygote were 206+/-11, 256+/-5, and 307+/-3 Hz, respectively. The fabricated biosensor was evaluated through an examination of 18 blind specimens. It could accurately discriminate between normal and SEA alpha-thalassemic samples, which suggests that this biosensor system is a promising alternative technique to detect SEA alpha-thalassemia because of its specificity and less hazardous exposure as compared with conventional methods.
Vattanaviboon
Phantip
P
Department of Clinical Microscopy, Faculty of Medical Technology, Mahidol University, Bangkok, Thailand. mtpvt@mahidol.ac.th
Sangseekhiow
Kulphassorn
K
Winichagoon
Pranee
P
Promptmas
Chamras
C
D001210:Asia, Southeastern; D015374:Biosensing Techniques / Q000295:instrumentation / Q000379:methods*; D004247:DNA / Q000032:analysis / Q000235:genetics; D004587:Electrophoresis, Agar Gel; D005914:Globins / Q000235:genetics; D006239:Haplotypes / Q000235:genetics*; D006801:Humans; D015345:Oligonucleotide Probes / Q000235:genetics*; D016133:Polymerase Chain Reaction / Q000379:methods*; D011791:Quartz; D015203:Reproducibility of Results; D017085:alpha-Thalassemia / Q000097:blood / Q000175:diagnosis / Q000235:genetics*
D015345:Oligonucleotide Probes; D011791:Quartz; D005914:Globins; D004247:DNA