Hande KURTULGAN KÜÇÜK, Öztürk ÖZDEMİR, Senol ÇİTLİ, Binnur KÖKSAL, Nejmiye AKKUŞ, İlhan SEZGİN

Prenatal dönemde multiplex kantitatif fluoresan PCR (QF-PCR) tekniği ile yaygın kromozomal anoploidilerin tespiti: Cumhuriyet Üniversitesi Tıp Fakültesi deneyimi

Amaç. Kantitatif fluoresan polimeraz zincir reaksiyonu (quantitative fluorescent polymerase chainreaction, QF-PCR) insanda major sayısal anoploidi nedeni olan kromozom 13, 18, 21, X ve Ykromozomlarının küçük ardıl tekrar (short tandem repeat, STR) analizi ile hızlıkimliklendirilmesine olanak sağlayan bir yöntemdir. Araştırmada, üçlü test pozitifliği ve diğernedenlerle riskli kabul edilen gebeliklerde QF-PCR tekniği kullanılarak fetal kromozomlarda STRanalizi yapılmış, yöntemin prenatal tanıdaki yeri ve önemi tartışılmıştır. Yöntem. Toplam 300 adetriskli gebelik analiz edilmiş olup analizlerin tamamında fetal amniyotik mayii (15 mL)kullanılmıştır. Bu amaçla Cumhuriyet Üniversitesi Tıp Fakültesi Kadın Hastalıkları ve DoğumAnabilim Dalı tarafından analiz için gönderilen fetal amniyotik mayiinin RPM1640 besi ortamındasolid hücre kültürü ile karyotip karyogram ve QF-PCR ile STR analizleri yapılmıştır. STR analiziçin yaklaşık 05-1 mL amniyotik mayii örneğinden izole edilen total genomik DNA kullanıldı.Fetal DNAlar uygun primerler kullanılarak kromozom 13, 18, 21, X ve Yye ait toplam 17 STRbölgesi fluoresan multiplex PCR yöntemiyle amplifiye edildi ve genotiplendirildi. Bulgular.Analiz için amniyotik mayii örneği gönderilen gebeliklerin 16-20 haftalık gebelikler oldukları veen önemli etiyolojik sebebin ise ileri anne yaşı olduğu tespit edildi. Sivas bölgesine aitlaboratuvarımıza gönderilen 300 adet amniyon mayii materyalinden 1 adet trizomi 13 (Patausendromu), 1 adet trizomi 18 (Edward sendromu), 5 adet trizomi 21 (Down sendromu) olmaküzere toplam 7 adet trizomik fetus tanısı kondu ve raporlandırıldı. Bu sonuçlar aynı zamandaamniyon hücre kültürü ile elde edilen karyotip analizi ile doğrulandı. Trizomik fetusların her ikiebeveynine fetus karyotipleri hakkında genetik danışma verildi. Sonuç. Prenatal tanıda QF-PCRyönteminin yararlılığı ortaya konmuş olup rutin prenatal tanıda güvenle kullanılabileceğigösterilmiştir.

Detection of common chromosome aneuploidies at the prenatal period by multiplex quantitative f luorescent PCR (QF-PCR) technique: The experience of pilot study in Cumhuriyet University of School of Medicine

Aim. Quantitative fluorescent polymerase chain reaction (QF-PCR) is a method that helps rapiddetection of chromosome 13, 18, 21, X and Y which are the major cause of numerical aneuploidiesin human using specific short tandem repeats (STR). In the current study, STR analysis, with usingQF-PCR technique, was done in fetal chromosomes of pregnancies which was accepted as risky interms of triple test positivity and other reasons and the value of the method in prenatal diagnosis.Method. A total of 300 risky fetus were analysed, fetal amniotic fluid (15 mL) was used for allanalyses. For this purpose, fetal amniotic fluids, which were referred by Cumhuriyet UniversitySchool of Medicine, Department of Obstetrics and Gynecology, were subjected to karyotyping bycell culture in RPM1640 culture medium and of-PCR STR analysis. Total genomic DNA samplesextracted from fetal amniotic fluid cells (1-1.5 mL) were used for STR analysis. Fetal DNAs wereamplified and then genotyped using appropriate primers belonging to chromosomes 13, 18, 21, Xand Y in terms of 17 STR region. Results. It was determined that the pregnant women whoseamniotic fluids were sent for analysis, were between 16-week and 20-week of pregnancies, and themost important etiologic reason was advanced mother age. A trisomy 13 (Patau syndrome), atrisomy 18 (Edwards Syndrome) and 5 trisomy 21 (Down syndrome) were detected in total ofamniotic fluids which were sent to our laboratory of Sivas region. These results were alsoconfirmed by cell culture and karyotype analysis. Genetic counselling about their fetuseskaryotype was given to both of the parents. Discussion. These results showed that QF-PCR is aneffective method and can be used safely in prenatal diagnosis.

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