Application of microsatellite loci on the chromosome X for rapid prenatal detection of the chromosome X numerical abnormalities

Abstract

AimTo determine the value of short-tandem repeat markers on the chromosome X (X-STR) for prenatal diagnostics of the chromosome X numerical disorders.MethodsWe investigated the genetic variability of 5 X-markers (DXS9895, DXS6810, DXS6803, GATA172D05, and HPRTB) in 183 healthy Croatian individuals (90 men and 93 women). We also tested 13 patients with X chromosome disorders (Turner syndrome – 6 cases; Klinefelter syndrome – 5 cases, and Triple X syndrome – 2 cases). The analysis was performed using polymerase chain reaction amplification with specific primers and electrophoresis on a polyacrylamide gel. The study was performed in 2010.ResultsOur sample showed no significant differences in allelic frequencies of the investigated X-markers from other European populations. A set of 5 X-STR markers was sufficiently informative for a successful determination of the chromosome X numerical abnormalities.ConclusionSince no false positive or negative results were observed, diagnostic value of the investigated X-STR loci for prenatal detection of chromosome X numerical disorders was confirmed. Our study represents an important step toward an improved prenatal diagnostics in Croatia.

Use of microsatellite loci in prenatal and postnatal diagnosis of aneuploidy and uniparental disomy

“…Iz 5 mL plodove vode i 200 μL krvi ispitanika izolirana je DNA komercijalnim setom za izolaciju (Nucleospin, Macherey Nagel, Duren, Njemačka). Uzorci humane genom ske DNA rabljeni su za umnažanje fragmenata točno željene veličine PCR-om te analizirani elektroforezom na 6%-tnom poliakrilamidnom gelu u automatskom sekven ceru (ALFexpress sekvencer, Pharmacia Biotech, Uppsala, Švedska) Korištena metoda je "in house" s početnicama opisanim u literaturi (8,9). Radi očitanja u automatskom sekvenceru sve 5' početni ce obilježene su fl uorescentnom bojom Cy-5 na svom 5' kraju.…”