Jamali, Somayeh, Mahdian, Reza, Hayat Nosaeid, Mina, Babashah, Sadegh, Maryami, Fereshteh, Karimipoor, Morteza, Zarbakhsh, Behnaz, Rahimi nejad, Faezeh, Zeinali, Sirous, Zeinali, Sirous (2008). Detection of unknown deletions in alpha globin genes in alpha thalassemia carriers using Real-time PCR. , 11(0), 31-39.
Somayeh Jamali; Reza Mahdian; Mina Hayat Nosaeid; Sadegh Babashah; Fereshteh Maryami; Morteza Karimipoor; Behnaz Zarbakhsh; Faezeh Rahimi nejad; Sirous Zeinali; Sirous Zeinali. "Detection of unknown deletions in alpha globin genes in alpha thalassemia carriers using Real-time PCR". , 11, 0, 2008, 31-39.
Jamali, Somayeh, Mahdian, Reza, Hayat Nosaeid, Mina, Babashah, Sadegh, Maryami, Fereshteh, Karimipoor, Morteza, Zarbakhsh, Behnaz, Rahimi nejad, Faezeh, Zeinali, Sirous, Zeinali, Sirous (2008). 'Detection of unknown deletions in alpha globin genes in alpha thalassemia carriers using Real-time PCR', , 11(0), pp. 31-39.
Jamali, Somayeh, Mahdian, Reza, Hayat Nosaeid, Mina, Babashah, Sadegh, Maryami, Fereshteh, Karimipoor, Morteza, Zarbakhsh, Behnaz, Rahimi nejad, Faezeh, Zeinali, Sirous, Zeinali, Sirous Detection of unknown deletions in alpha globin genes in alpha thalassemia carriers using Real-time PCR. , 2008; 11(0): 31-39.

Detection of unknown deletions in alpha globin genes in alpha thalassemia carriers using Real-time PCR

Pathobiology Reserach
Article 5, Volume 11, Issue 0, 2008, Pages 31-39    PDF (619.85 K)
Authors
Somayeh Jamali1; Reza Mahdian1; Mina Hayat Nosaeid1; Sadegh Babashah2; Fereshteh Maryami1; Morteza Karimipoor1; Behnaz Zarbakhsh1; Faezeh Rahimi nejad3; Sirous Zeinali* 4; Sirous Zeinali* 4
1Department of Molecular Medicine, Pasteur Institute of Iran, Tehran, Iran
2Department of Genetics, Science and Research Branch of Islamic Azad University, Tehran, Iran
3Kawsar Human Genetics Researech Center, Tehran, Iran
4Department of Genetics, Kawsar Human Genetics Research Center, Tehran, Iran
Abstract
Objective: Alpha-thalassemia is one of the most prevalent hemoglobin disorders in the world and it is a common hereditary condition caused by deletion of one or more α-globin genes. Common α-thalassemia deletions like 3.7 kb, 4.2 kb, 20.5 kb and Med can be detected by Multiplex PCR. There are, however, some unknown deletions that can not be detected by the mentioned method or even by direct DNA sequencing. In the present study, Real-time PCR was used to determine the presence or absence of unknown deletions.
Materials and Methods: Real-time PCR was performed using intercalating dye SYBR Green I and α1, α2 and CLCN7 genes were amplified. Data analysis was conducted using comparative threshold method (ΔΔCT) for determination of Gene dosage of α1-globin and α2-globin genes.
Results: The results showed the ratio of 0.90±0.16 for normal individuals and the ratio of 0.32±0.15 for carrier samples with deletions. In addition, Melting curve analysis confirmed the specific amplification of target genes.
Conclusion: The Real-time PCR assay is simple, rapid, and reliable. It can be applied for direct determination of unknown deletions in Alpha-thalassemia carriers.
Keywords
Real-time PCR; α-thalassemia; Threshold cycle
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