Direct detection of fetal cells in maternal blood: a reappraisal using a combination of two different Y chromosome-specific FISH probes and a single X chromosome-specific probe

Background: We have recently explored the detection of circulatory male fetal cells directly in maternal whole blood samples by fluorescence in-situ hybridization (FISH). In order to improve the efficacy of fetal cell detection, we have now examined whether this could be enhanced by the use of two different Y chromosome-specific FISH probes (α-satellite and classical satellite III regions) in combination with an X chromosome-specific FISH probe. Methods: Nineteen maternal blood samples (median gestational age=28weeks, range=12-37weeks) were examined in a blinded manner. No enrichment procedure was performed. Following hypotonic treatment and Carnoy's fixation, total nucleated cells were examined by two color FISH with a single X and two Y chromosome-specific probes. Nine cases were examined in parallel by conventional XY-FISH. Results: Fetal cell detection was superior when using two Y chromosome-specific probes (specificity=75%; sensitivity=91%) when compared to the conventional XY-FISH approach (specificity=50%; sensitivity=60%). Conclusions: Male fetal cells can be detected in most maternal blood samples examined. Specificity and sensitivity is improved when using a combination of single X and two Y chromosome-specific probes when compared to a conventional XY-FISH protoco

Persönlichkeitsmerkmale von Schweinefachberatern und Tierärzten in einer gemeinschaftlichen Tiergesundheitsberatung

Eine gemeinschaftlich ausgerichtete Tiergesundheitsberatung für schweinehaltende Betriebe durch produktionstechnische Berater und Hoftierärzte kann Vorteile bieten. Ziel der vorliegenden Untersuchungen ist es, den Zusammenhang zwischen Persönlichkeitsmerkmalen nach dem Big-Five-Modell und dem Grad der Zusammenarbeit zu analysieren. Die vorliegende Untersuchung umfasst Daten einer standardisierten persönlichen Befragung von 24 schweinehaltenden Betrieben. Gemeinsame Beratung ist tendenziell erfolgreicher. Eher extravertierte Personen sowie stärker verträgliche Tierärzte und weniger verträgliche Berater sind für eine gemeinsame Beratung eher geeignet. Die vorliegenden Ergebnisse weisen darauf hin, dass neben den oftmals analysierten technischen, auch sozial-organisatorische Innovationen in einem ganzheitlichen Ansatz im Tiergesundheitsmanagement beachtet werden sollte

Isolation of cell-free DNA from maternal plasma using manual and automated systems

Cell-free fetal DNA present in the maternal circulation holds great potential for noninvasive prenatal diagnosis and analysis of fetal genetic traits. However, only approximately 3-6% of total DNA in the maternal plasma is of fetal origin. Because of its scarcity in the maternal circulation, various methods have been developed and tested to optimize the extraction of this rare material from plasma. Here, we first describe the commonly used protocol for separating plasma from whole blood samples. We also describe two commercially available methods for the extraction of cell-free DNA from maternal plasma, which we have found particularly straightforward and easy to use: a manual method using the High Pure PCR Template Preparation kit (Roche Diagnostics) and an automated system using the MagNA Pure LC instrument (Roche Diagnostics). Use of the methods described here will help to ensure maximum yield and purity of cell-free fetal DNA extracted from maternal plasma samples for downstream analyses