科研之友
微信
新浪微博
Facebook
分享链接
摘要
We used quantitative real-time polymerase chain reaction (qPCR) and high resolution melting (HRM) analyses for the detection of the common alpha-thalassemia (alpha-thal) genotypes in 40 northern Thai Hb H (beta 4) patients. The alpha(0)-thal [--(SEA) (Southeast Asian) deletion] was detected by a multiplex gap real-time PCR. To determine the alpha(1)-thal, three primer pairs were designed. The A-primer pair was used to amplify the 3%26apos; terminal DNA sequences of the HBA2 gene, and the B-primer pair amplified the 5%26apos; flanking region of the HBA1 gene. The C-primer pair amplified the 3%26apos; terminal DNA sequences of the HBA1 gene and was used as an internal control. The -alpha(4.2) (leftward) and -alpha(3.7) (rightward) deletions were determined by monitoring the absence of PCR product(s). The Hb H patients who had a negative PCR result for the A-primer pair but positive for the B- and C-primer pairs carried the -alpha 4.2 deletion, while the -alpha 3.7 deletion carriers were negative for the A- and B-primer pairs. In the case of Hb H with Hb Constant Spring (Hb CS, alpha 142, Term -%26gt; Gln; HBA2: c. 427T%26gt;C), all primer pairs were positive, HRM analysis of the PCR product of the A-primer pair was introduced to analyze the Hb CS gene. It can distinguish clearly between normal and Hb CS genotypes. The applied methods for the determination of the alpha(0)- and alpha(+)-thal genotypes revealed the results to be as accurate as conventional gap-PCR and the direct DNA sequencing methods but resulted in a much simpler and faster procedure.
- 出版日期2013
