بهينه سازي و كاربرد تكنيك Real-Time PCR كمي با استفاده از SYBR-Green 1 در تشخيص سندرم داون

Development and Application of Real-Time Quantitative Polymerase Chain Reaction Technique Using SYBR Green I in the Diagnosis of Down Syndrome

Objective: Rapid diagnosis of Trisomy 21 Syndrome (Down Syndrome) patients using Real-Time quantitative Polymerase Chain Reaction (Real-Time qPCR) in order to establish a novel method for prenatal diagnosis in the future. Materials and Methods: A total of 5 ml of peripheral blood was obtained from each patient and normal controls (NR). Then, genomic DNA from lymphocytes was extracted using the salting out procedure. Gene dosage levels of DSCAM and (PMP22, DSCAM) in Down Syndrome and NR were analyzed using real-time quantitative PCR. The DSCAM/ PMP22 ratio was calculated according to the 2 -AACt formula for all samples. Results: Real-time PCR showed a DSCAM/PMP22 ratio of 1.48 ± 0.18 and 1.01 ± 0.10 (p<0.001) in Down Syndrome and normal samples, respectively, demonstrating three copies of the target (DSCAM) gene in Trisomy 21 Syndrome. Conclusion: DSCAM/PMP22 ratio is increased significantly in Down Syndrome patients than NR (1.5 times). Therefore, the real-time quantitative PCR technique can be used as a sensitive, accurate and reliable technique for rapid and prenatal diagnosis of Trisomy 21 Syndrome.