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Sperm Chromatin Structure Assay (SCSA®)

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Spermatogenesis

Part of the book series: Methods in Molecular Biology ((MIMB,volume 927))

Abstract

The SCSA® is the pioneering assay for the detection of damaged sperm DNA and altered proteins in sperm nuclei via flow cytometry of acridine orange (AO) stained sperm. The SCSA® is considered to be the most precise and repeatable test providing very unique, dual parameter data (red vs. green fluorescence) on a 1,024 × 1,024 channel scale, not only on DNA fragmentation but also on abnormal sperm characterized by lack of normal exchange of histones to protamines. Raw semen/sperm aliquots or purified sperm can be flash frozen, placed in a box with dry ice and shipped by overnight courier to an experienced SCSA® lab. The samples are individually thawed, prepared, and analyzed in ∼10 min. Of significance, data on 5,000 individual sperm are recorded on a 1,024 × 1,024 dot plot of green (native DNA) and red (broken DNA) fluorescence. Repeat measurements have virtually identical dot plot patterns demonstrating that the low pH treatment that opens up the DNA strands at the sites of breaks and staining by acridine orange (AO) are highly precise and repeatable (CVs of 1–3%) and the same between fresh and frozen samples. SCSAsoft® software transforms the X-Y data to total DNA stainability versus red/red + green fluoresence (DFI) providing a more accurate determination of % DFI as well as the more sensitive value of standard deviation of DFI (SD DFI) as demonstrated by animal fertility and dose–response toxicology studies. The current established clinical threshold is 25% DFI for placing a man into a statistical probability of the following: (a) longer time to natural pregnancy, (b) low odds of IUI pregnancy, (c) more miscarriages, or (d) no pregnancy. Changes in lifestyle as well as medical intervention can lower the %DFI to increase the probability of natural pregnancy. Couples of men with >25% DFI are counseled to try ICSI and when in the >50% range may consider TESE/ICSI. The SCSA® simultaneously determines the % of sperm with high DNA stainability (%HDS) related to retained nuclear histones consistent with immature sperm; high HDS values are predictive of pregnancy failure.

The SCSA® is considered to be the most technician friendly, time- and cost-efficient, precise and repeatable DNA fragmentation assay, with the most data and the only fragmentation assay with an accepted clinical threshold for placing a man at risk for infertility. SCSA® data are more predictive of male factor infertility than classical semen analyses.

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Author information

Authors and Affiliations

  1. SCSA Diagnostics, Volga, SD, USA

    Donald P. Evenson

  2. Department of Obstetrics and Gynecology, Emeritus, South Dakota State University, Sanford Medical School, and University of South Dakota, Vermillion, SD, USA

    Donald P. Evenson

Authors
  1. Donald P. Evenson
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Correspondence to Donald P. Evenson .

Editor information

Editors and Affiliations

  1. Andrology and IVF Laboratories, Department of Surgery (Urology), University of Utah School of Medicine, Arapeen Dr. 675, Salt Lake City, 84108, Utah, USA

    Douglas T. Carrell

  2. , Andrology and IVF Laboratories, University of Utah School of Medicine, Arapeen Way 675, Salt Lake City, 84108, Utah, USA

    Kenneth I. Aston

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Evenson, D.P. (2013). Sperm Chromatin Structure Assay (SCSA®). In: Carrell, D., Aston, K. (eds) Spermatogenesis. Methods in Molecular Biology, vol 927. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-62703-038-0_14

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  • DOI: https://doi.org/10.1007/978-1-62703-038-0_14

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  • Publisher Name: Humana Press, Totowa, NJ

  • Print ISBN: 978-1-62703-037-3

  • Online ISBN: 978-1-62703-038-0

  • eBook Packages: Springer Protocols

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