Accurate chromosome testing of miscarriages with manual vacuum aspiration (MVA) and DNA technologies: overcoming culture failure and maternal cell contamination (MCC) from cultured decidua

Accurate chromosome testing of miscarriages with manual vacuum aspiration (MVA) and DNA... OBJECTIVE: To assess the use of MVA and DNA technologies for overcoming pitfalls associated with chromosome testing of miscarriages. DESIGN: Prospective cohort. MATERIALS AND METHODS: With IRB approval, women with ≥2 miscarriages of <10 wks, seen between 06/2004-07/2008, were identified. Prior to consultation, chromosome testing of miscarriages consisted of cytogenetic analysis only. Subsequently, pregnancy was defined as a +ve hCG drawn at 4 wks. Miscarriage tissue was obtained by office MVA, isolated by the clinician and sent for cytogenetic analysis. With 46,XX, microsatellite testing was performed, which compared highly polymorphic loci in the miscarriage DNA to the woman's DNA. If MCC was confirmed, comparative genomic hybridization (CGH) was performed on cryopreserved miscarriage tissue. Fisher's exact test and 95% confidence intervals for binomial proportions were reported. RESULTS: Success rates (livebirths and ongoing pregnancies >14 wks) prior and subsequent to consultation were 12% (187/1559) and 58% (195/337), respectively. Table 1 Prior Subsequent RPL subjects 368 251 Mean age at miscarriage 33 yrs (16-45) 36 yrs (27-46) Miscarriages <10 wks 1173 122 Miscarriages sent for chromosome testing 18% (211/1173) 55% (67/122) Culture failure 9% (19/211) 1% (1/67) p=0.05 MCC Unknown 20% (3/15) ∗ Aneuploid/Polyploid 37.5% (72/192) 50.8% (31/61) Euploid 62.5% (120/192) 49.2% (30/61) Ratio of 46,XX / 46,XY 2.2 (82/38) 1.1 (16/14) Proportion of 46,XX 68% (82/120) (59, 77%) 53% (16/30) (34, 72%) ∗ 15 of 19 46,XX miscarriages available for testing CONCLUSIONS: Prior to consultation, few miscarriages were sent for chromosome testing. Of those, there was a disproportionate number of 46,XX results, most likely due to MCC. With MVA, isolation of the miscarriage tissue, and DNA technologies when indicated, the 46,XX/46XY ratio normalized. Accurate chromosome testing of miscarriages requires rigorous handling techniques, conventional cytogenetic analysis and the judicious use of DNA technologies.</P> http://www.deepdyve.com/assets/images/DeepDyve-Logo-lg.png Fertility and Sterility Elsevier

Accurate chromosome testing of miscarriages with manual vacuum aspiration (MVA) and DNA technologies: overcoming culture failure and maternal cell contamination (MCC) from cultured decidua

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Publisher
Elsevier
Copyright
Copyright © 2009 American Society for Reproductive Medicine
ISSN
0015-0282
DOI
10.1016/j.fertnstert.2009.07.1027
Publisher site
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Abstract

OBJECTIVE: To assess the use of MVA and DNA technologies for overcoming pitfalls associated with chromosome testing of miscarriages. DESIGN: Prospective cohort. MATERIALS AND METHODS: With IRB approval, women with ≥2 miscarriages of <10 wks, seen between 06/2004-07/2008, were identified. Prior to consultation, chromosome testing of miscarriages consisted of cytogenetic analysis only. Subsequently, pregnancy was defined as a +ve hCG drawn at 4 wks. Miscarriage tissue was obtained by office MVA, isolated by the clinician and sent for cytogenetic analysis. With 46,XX, microsatellite testing was performed, which compared highly polymorphic loci in the miscarriage DNA to the woman's DNA. If MCC was confirmed, comparative genomic hybridization (CGH) was performed on cryopreserved miscarriage tissue. Fisher's exact test and 95% confidence intervals for binomial proportions were reported. RESULTS: Success rates (livebirths and ongoing pregnancies >14 wks) prior and subsequent to consultation were 12% (187/1559) and 58% (195/337), respectively. Table 1 Prior Subsequent RPL subjects 368 251 Mean age at miscarriage 33 yrs (16-45) 36 yrs (27-46) Miscarriages <10 wks 1173 122 Miscarriages sent for chromosome testing 18% (211/1173) 55% (67/122) Culture failure 9% (19/211) 1% (1/67) p=0.05 MCC Unknown 20% (3/15) ∗ Aneuploid/Polyploid 37.5% (72/192) 50.8% (31/61) Euploid 62.5% (120/192) 49.2% (30/61) Ratio of 46,XX / 46,XY 2.2 (82/38) 1.1 (16/14) Proportion of 46,XX 68% (82/120) (59, 77%) 53% (16/30) (34, 72%) ∗ 15 of 19 46,XX miscarriages available for testing CONCLUSIONS: Prior to consultation, few miscarriages were sent for chromosome testing. Of those, there was a disproportionate number of 46,XX results, most likely due to MCC. With MVA, isolation of the miscarriage tissue, and DNA technologies when indicated, the 46,XX/46XY ratio normalized. Accurate chromosome testing of miscarriages requires rigorous handling techniques, conventional cytogenetic analysis and the judicious use of DNA technologies.</P>

Journal

Fertility and SterilityElsevier

Published: Sep 1, 2009

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