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Journal of Family and Reproductive Health، جلد ۱۱، شماره ۳، صفحات ۱۴۶-۱۵۱
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عنوان انگلیسی Comparing the Efficiency of Three Protocols in Isolation of Cell Free Fetal DNA From Maternal Blood
چکیده انگلیسی مقاله Objective: Recent advances in non-invasive prenatal diagnosis (NIPD) through cell free fetal DNA (cffDNA) has highlighted cffDNA purification as a critical initial step. Herein, we aimed to compare the efficiency of one proposed protocol with two commercial kits for isolation of cffDNA. Materials and methods: cffDNA was isolated from whole blood of 50 normal pregnancies using one proposed manual protocol compared with QIAamp DNA Blood Mini and Bioneer Kit s. Methylated DNA immunoprecipitation real time polymerase chain reaction (MeDIP-Real time PCR) was performed to quantify three fetal specific sequences. Results: Maximum cffDNA quantity was obtained by suggested protocol (248.79 ± 14.07 ng/µl) and the best quality was achieved by Bioneer Kit (OD ratio: 260/280 nm/nm: 1.69 ± 0.09, 260/230 nm/nm: 1.15 ± 0.13) (p < 0.001). Enrichment of fetal specific sequences was significantly higher when proposed protocol was used to isolate cffDNA (p = 0.01). Conclusion: Inhibitory effect of NaI on nucleases and double digestion of DNA associated proteins may be the main reasons behind the superiority of suggested protocol. Significantly higher amplification of fetal specific sequences in suggested protocol would be a strong evidence on recovery of small fetal fragments as demonstrated with its maximum total DNA quantity and amplification in different PCR reactions.
کلیدواژه‌های انگلیسی مقاله DNA,Isolation,NIPD,Maternal Blood,Protocol
نویسندگان مقاله | Fatemeh Karami
Department of Medical Genetics, School of Medicine, Tehran University of Medical Sciences, Tehran, Iran


| Mohammad Reza Noori-Daloii
Vali-e-Asr Reproductive Health Research Center, School of Medicine, Tehran University of Medical Sciences, Iran


| Seddigheh Hantooshzadeh
Departement of Medical Genetics, Science and Research Branch, Islamic Azad University, Tehran, Iran; Department of Medical Genetics, School of Medicine, Tehran University of Medical Sciences, Tehran, Iran


| Mohammad Hossein Modarressi


نشانی اینترنتی http://jfrh.tums.ac.ir/index.php/jfrh/article/view/652
فایل مقاله اشکال در دسترسی به فایل - ./files/site1/rds_journals/186/article-186-575586.pdf
کد مقاله (doi)
زبان مقاله منتشر شده en
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نوع مقاله منتشر شده Original Articles
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