สถาบันวิจัยวิทยาศาสตร์สาธารณสุข

National Institute of Health of Thailand

Authors : Somchai Sangkitporn*, Siripakorn Sangkitporn*, Areerat Sangnoi*, Prissana Charoenporn*.

 

Affiliations:       *National Institute of Health, Department of Medical Sciences, Nonthaburi 11000
 
Source:            Journal of Allied Health Sciences 2003; 3: 207-214
 
Language:        Thai with English abstract
 
Abstract:
 
A method for diagnosis of a0-thalassemia was developed based on detection of accumulated PCR product using SYBR Green I, a double-stranded DNA binding dye, and a fluorescence-detecting thermocycler. Primers were designed to specifically amplify - -SEA and      - -Thai deletions of a0-thalassemia. Albumin was selected as the reference gene. The comparative CT method was used to quantitate the target gene relative to the albumin gene. Dissociation curve analysis was used as a qualitative tool distinguish different types of a0-thalassemia. In this study, the melting temperatures of the - -Thai and - -SEA deletions were 83 and 86oC, respectively. Application of the assay for the diagnosis of a0-thalassemia heterozygotes and homozygotes is reported. The assay is highly robust and amenable to high throughput. It is thus a useful tool for the rapid detection of a0-thalassemia in populations with a high frequency of a0-thalassemia,       - -Thai and - -SEA deletions