PMID- 26919909 OWN - NLM STAT- MEDLINE DCOM- 20170119 LR - 20220408 IS - 1873-2232 (Electronic) IS - 0378-4320 (Linking) VI - 169 DP - 2016 Jun TI - The Sperm Chromatin Structure Assay (SCSA((R))) and other sperm DNA fragmentation tests for evaluation of sperm nuclear DNA integrity as related to fertility. PG - 56-75 LID - S0378-4320(16)30027-6 [pii] LID - 10.1016/j.anireprosci.2016.01.017 [doi] AB - Thirty-five years ago the pioneering paper in Science (240:1131) on the relationship between sperm DNA integrity and pregnancy outcome was featured as the cover issue showing a fluorescence photomicrograph of red and green stained sperm. The flow cytometry data showed a very significant difference in sperm DNA integrity between fertile and subfertile bulls and men. This study utilized heat (100 degrees C, 5min) to denature DNA at sites of DNA strand breaks followed by staining with acridine orange (AO) and measurements of 5000 individual sperm of green double strand (ds) DNA and red single strand (ss) DNA fluorescence. Later, the heat protocol was changed to a low pH protocol to denature the DNA at sites of strand breaks; the heat and acid procedures produced the same results. SCSA data are very advantageously dual parameter with 1024 channels (degrees) of both red and green fluorescence. Hundreds of publications on the use of the SCSA test in animals and humans have validated the SCSA as a highly useful test for determining male breeding soundness. The SCSA test is a rapid, non-biased flow cytometer machine measurement providing robust statistical data with exceptional precision and repeatability. Many genotoxic experiments showed excellent dose response data with very low coefficient of variation that further validated the SCSA as being a highly powerful assay for sperm DNA integrity. Twelve years following the introduction of the SCSA test, the terminal deoxynucleotidyl transferase-mediated fluorescein-dUTP nick end labelling (TUNEL) test (1993) for sperm was introduced as the only other flow cytometric assay for sperm DNA fragmentation. However, the TUNEL test can also be done by light microscopy with much less statistical robustness. The COMET (1998) and Sperm Chromatin Dispersion (SCD; HALO) (2003) tests were introduced as light microscope tests that don't require a flow cytometer. Since these tests measure only 50-200 sperm per sample, they suffer from the lack of the statistical robustness of flow cytometric measurements. Only the SCSA test has an exact standardization of a fixed protocol. The many variations of the other tests make it very difficult to compare data and thresholds for risk of male factor infertility. Data from these four sperm DNA fragmentation tests plus the light microscope acridine orange test (AOT) are correlated to various degrees. CI - Copyright (c) 2016 The Authors. Published by Elsevier B.V. All rights reserved. FAU - Evenson, Donald P AU - Evenson DP AD - SCSA Diagnostics, Inc., Brookings, SD, United States; South Dakota State University, Brookings, SD, United States; University of South Dakota, Sanford Medical School, Dept. OB/GYN, Sioux Falls, SD, United States. Electronic address: don@scsatest.com. LA - eng PT - Journal Article DEP - 20160217 PL - Netherlands TA - Anim Reprod Sci JT - Animal reproduction science JID - 7807205 RN - 0 (Chromatin) SB - IM MH - Animals MH - *Chromatin MH - *DNA Fragmentation MH - Female MH - Flow Cytometry/methods MH - Male MH - Pregnancy MH - Pregnancy Outcome MH - Semen Analysis/methods MH - Spermatozoa/*cytology OTO - NOTNLM OT - Fragmentation tests OT - Large and small animals OT - Male fertility OT - Other sperm DNA OT - Sperm Chromatin Structure Assay (SCSA) OT - Toxicology EDAT- 2016/02/28 06:00 MHDA- 2017/01/20 06:00 CRDT- 2016/02/28 06:00 PHST- 2015/12/11 00:00 [received] PHST- 2016/01/13 00:00 [revised] PHST- 2016/01/25 00:00 [accepted] PHST- 2016/02/28 06:00 [entrez] PHST- 2016/02/28 06:00 [pubmed] PHST- 2017/01/20 06:00 [medline] AID - S0378-4320(16)30027-6 [pii] AID - 10.1016/j.anireprosci.2016.01.017 [doi] PST - ppublish SO - Anim Reprod Sci. 2016 Jun;169:56-75. doi: 10.1016/j.anireprosci.2016.01.017. Epub 2016 Feb 17.