1. Rapid comparative genomic hybridization protocol for prenatal diagnosis and its application to aneuploidy screening of human polar bodies 
Christina Landwehr, M.Sc., Markus Montag, Ph.D., Katrin van der Ven, M.D., Ruthild G. Weber, M.D. 
Fertility and Sterility 
Volume 90, Issue 3, Pages (September 2008)
DOI: /j.fertnstert
Copyright © 2008 American Society for Reproductive Medicine Terms and Conditions

2. Figure 1
Establishing and validating the rapid CGH protocol. In all CGH profiles shown, the centromeres of all chromosomes, the heterochromatin block on chromosome 1, as well as the short arms of the acrocentric chromosomes (13, 21, 22) were shaded to indicate regions of tandem repetitive DNA sequences, which were excluded from the CGH evaluation. (A) Chromosomal CGH profiles obtained by using the standard CGH protocol (top row) and the rapid CGH protocol (bottom row) when analyzing DNA from our test system, the ovarian adenocarcinoma cell line COLO 704, carrying gains of entire large and small chromosomes as well as a gain of a chromosome arm. Gains of chromosomes and chromosomal arms 1q, 7, 8, 19, and 20 that were detected by the standard CGH protocol (top) are also diagnostic when using rapid CGH (bottom). (B) Top, trisomy 18 is detected in the karyotype of a fibroblast from a tendon biopsy of an aborted fetus by GTG banding (top left), and a gain of chromosome 18 is found when analyzing a single fibroblast from the same tendon biopsy culture by using rapid CGH (top right). Bottom, trisomy 21 is detected in the karyotype of a lymphocyte from a patient with Down's syndrome by GTG banding (bottom left), and a gain of chromosome 21 is found when analyzing a single lymphocyte from the same blood culture by using rapid CGH (bottom right). (C) Top, analysis of lymphocyte nuclei with trisomy 21 from a patient with Down's syndrome by using interphase FISH and rapid CGH. Top left, FISH analysis of a lymphocyte nucleus showing three signals for chromosome 21 (red, with filled arrowheads) and two regular signals for chromosome 13 (green, with open arrowheads). Top right, another lymphocyte nucleus from the same glass slide was then microdissected, and rapid CGH was performed and showed a shift of the chromosome 21 profile to the right, equivalent to a chromosome 21 gain, whereas the profile for chromosome 13 was balanced. Bottom, consecutive analysis of the same first polar body with an extra chromatid of chromosome 22 by using interphase FISH and rapid CGH. Bottom left, polar body analyzed by FISH showing three signals for chromosome 22 (green, with open arrowheads), equivalent to an extra chromatid, and two regular signals for chromosome 18 (red, with filled arrowheads). Bottom right, the polar body was then microdissected from the glass slide, and rapid CGH was performed and showed a gain of chromosome 22, whereas the profile for chromosome 18 was balanced. (D) Identification of extra chromosomal material on a derivative chromosome 13 from a patient with mental retardation by using rapid CGH, with verification by FISH. Left, GTG banding showing a normal chromosome 13 and a derivative chromosome 13 containing extra material in the short arm. Center, rapid CGH profile of chromosome 13 demonstrating a partial gain of distal 13q. The additional material attached to the short arm of the derivative chromosome 13 is thus identified as material originating from the distal long arm of chromosome 13. Right, the rapid CGH result is verified by FISH analysis with clones RP11-120L14 (red, located at 13q31.1) and RP11-430M15 (green, located at 13q33.1), showing signals on the short arm of the derivative chromosome 13, in addition to signals at the expected bands of the long arm of chromosome 13.
Fertility and Sterility  , DOI: ( /j.fertnstert )
Copyright © 2008 American Society for Reproductive Medicine Terms and Conditions

3. Figure 2
Applying the rapid CGH protocol to aneuploidy screening of first polar bodies. In all CGH profiles shown, the centromeres of chromosomes were shaded to indicate regions of tandem repetitive DNA sequences, which were excluded from the CGH evaluation. (A) Frequency of chromosomal imbalances detected in 32 first polar bodies from 16 patients aged 33 to 44 years (average age, 38 y) who were undergoing infertility treatment mainly for AMA or RIF. Gains (black upward columns) and/or losses (light-gray downward columns) were detected in 75% of polar bodies, involving almost all chromosomes at least once. Among the detected aberrations in polar bodies, 40% were gains resulting in monosomies in the fertilized oocytes. (B) Comparative genomic hybridization profiles obtained after rapid CGH analysis of polar bodies from two selected cases. Top row, CGH analysis of polar body PB8A from a 39-year-old patient treated for AMA showing multiple aneuploidies (losses of chromosomes 6, 7, 8, 11, and 19). Bottom row, CGH analysis of polar body PB15B from a 41-year-old patient treated for AMA. No imbalances of any chromosome were detected. The profiles from the same chromosomes as in the top row are shown for comparison. A pregnancy resulted after implantation of the corresponding fertilized oocyte.
Fertility and Sterility  , DOI: ( /j.fertnstert )
Copyright © 2008 American Society for Reproductive Medicine Terms and Conditions