Application Guide - QIAxcel Advanced

52 QIAxcel Advanced Application Guide 10/2016 Rapid and accurate detection of Y chromosome microdeletions Natasa Teran,1 Mirjana Kozulic,2 and Borut Peterlin1 1 University Medical Centre Ljubljana, Ljubljana, Slovenia 2 QIAGEN Instruments AG, Hombrechtikon, Switzerland Y chromosome microdeletions and rearrangements that are relatively common causes of aberrant sperm profiles were detected using the QIAxcel® system. Samples were analyzed directly without prior manipulation, and the type of deletion was detected by automatic fragment-size determination. Use of the QIAxcel system significantly accelerated research on male infertility. Introduction The 3 azoospermia factor (AZF) regions of the Y chromosome accommodate genes required for spermatogenesis. The most distal region, AZFc, is one of the most genetically dynamic regions in the human genome (1, 2). Some AZF rearrangements are responsible for marked spermatogenic defects. For example, men with deletions in AZF regions exhibit drastically reduced sperm concentrations (<1 million sperm/ml compared to normal levels of >20 million sperm/ml) (3). A rapid and reproducible method for determining AZF deletions would support effective research on the mechanisms of Y chromosome microdeletions. To develop an optimal screening strategy for male subfertility, a large database of microdeletion data was constructed and analyzed (4, 5). Most of the published Y chromosome microdeletions (85.6%) can be detected by PCR using a set of 6 genetic markers: sY84, sY127, sY152, RBM1, sY147 and sY254-DAZ as well as for the ZFY/ZFX gene as an internal control for the multiplex PCR reaction (5). The QIAxcel system.

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