Paper Information

Journal:   ARCHIVES OF IRANIAN MEDICINE   APRIL 2013 , Volume 16 , Number 4; Page(s) 217 To 220.
 
Paper: 

COMPARISONS BETWEEN RT-PCR, REAL-TIME PCR, AND IN VITRO GLOBIN CHAIN SYNTHESIS BY α/β RATIO CALCULATION FOR DIAGNOSIS OF α- FROM β-THALASSEMIA CARRIERS

 
 
Author(s):  MARYAMI FERESHTEH, MAHDIAN REZA, JAMALI SOMAYEH, KARIMI ARZANANI MOHSEN, KHATAMI SHOHREH, MARYAMI FAHIMEH, BAYAT PARASTOU, SADEGHI SEDIGHE, KARIMIPOUR MORTEZA, ZEINALI SIROUS*
 
* BIOTECHNOLOGY RESEARCH CENTER, DEPARTMENT OF MOLECULAR MEDICINE, PASTEUR INSTITUTE OF IRAN, TEHRAN, IRAN
 
Abstract: 

Background: Thalassemia, which may be due to point mutations, translocations, and deletions involving the a or b globin gene, is the most prevalent single gene disorder in Iran. This study aims to calculate the a/b ratio in normal cases, a- and b-thalassemia carriers by RT-PCR, real-time PCR, and in vitro globin chain synthesis (GCS) in order to establish the most accurate technique to distinguish between a- and b-thalassemia carriers in suspicious cases.
Methods: The
a/b ratios were calculated in all samples by RT-PCR, real-time RT-PCR, and in vitro GCS.
Results: Using RT-PCR, the ratios were 1.09±0.07 in normal samples, 1.2±0.17 in
b-thalassemia, 1.08 ±0.19 in mild a-thalassemia, and 0.96±0.19 in severe a-thalassemia carriers. In real-time RT-PCR, the ratios were 2.21±1.36 in normal samples, 5.12±1.83 in b-thalassemia, 2.88±0.81 in mild a-thalassemia, and 1.18±0.52 in severe a-thalassemia carriers. With GCS, the ratios were 1.03±0.1 in normal samples, 1.9±0.37 in b-thalassemia, 0.8±0.13 in mild a-thalassemia, and 0.59±0.12 in severe a-thalassemia carriers.
Conclusion: To determine the most accurate technique, we statistically analyzed the
a/b ratios obtained from the three standard methods.
The ratio obtained by GCS and real-time PCR were helpful in distinguishing between
a and b carriers in suspicious patients in whom the mutation detection was limited and the risk for offspring was not clear. The use of this technique is more obvious when time is restricted (i.e. during the pregnancy period).

 
Keyword(s): COMPARATIVE THRESHOLD CYCLE METHOD, GLOBIN CHAIN SYNTHESIS, REAL-TIME PCR, RT-PCR, THALASSEMIA
 
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