Cited by 1,072Sperm and Testicular FunctionReproductive biology and impacts on aquatic speciesMale Reproductive Health Studies

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Sperm Chromatin Structure Assay: Its Clinical Use for Detecting Sperm DNA Fragmentation in Male Infertility and Comparisons With Other Techniques

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EVENSON, DONALD P., LARSON, KJERSTEN L., JOST, LORNA K.·2002·Journal of Andrology

DOI 10.1002/j.1939-4640.2002.tb02599.x

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Abstract

The Sperm Chromatin Structure Assay (SCSA) was first described in the December 5, 1980 issue of Science (Evenson et al, 1980a).The data described in that article showed a significant difference between proven fertile and subfertility or infertility in men and bulls and the susceptibility to denaturation of their sperm nuclear DNA.A subsequent study (Evenson et al, 1984) described SCSA data obtained from men with testicular cancer.The data were heterogeneous, and begged the question, what was SCSA measuring?Great effort has been spent over the past 15 years on animal model systems, mostly related to dose-response toxicology experiments and large animal fertility trials.The results from these studies showed that SCSA was highly dose-responsive to toxicants, highly repeatable, and provided meaningful biological information on sperm nuclear DNA defects.The extensive data from nonhumans (Evenson et al, 1985(Evenson et al, , 1986(Evenson et al, , 1989a(Evenson et al, ,b, 1993a(Evenson et al, ,b,c, 1994(Evenson et al, , 1995b;; Ballachey et

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Fertilization and pregnancy after assisted oocyte activation and intracytoplasmic sperm injection in a case of round-headed sperm associated with deficient oocyte activation capacity
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Longitudinal Assessment of Semen Quality Following SARS-CoV-2 Infection
Sadeghi, N., Ghorban, Z., Tavalaee, M., & Nasr-Esfahani, M. H. (2026) · Journal of Arak University of Medical Sciences · doi:10.66224/jams.29.1.70
Effects of High-dose Radioactive Iodine Therapy on Hormonal Profiles and Sperm Quality in Thyroidectomy Patients
Alavi, M., Hojjat, R., Taghinezhad, A., & Shojaeifard, M. (2026) · Molecular Imaging and Radionuclide Therapy · doi:10.4274/mirt.galenos.2025.90692
“The assay measures the susceptibility of sperm DNA to acid-induced denaturation in situ , followed by staining with the fluorescent dye acridine orange (17,18).”
Sperm DNA fragmentation in mammals: etiology, detection, fertility assessment under radiation exposure
Starobor, N., & Raskosha, O. (2025) · Proceedings of the Komi Science Centre of the Ural Division of the Russian Academy of Sciences · doi:10.19110/1994-5655-2025-7-35-49
Imiquimod (R837), a TLR7-Specific Agonist, Regulates Boar Sperm Motility via PI3K/GSK3α/β/Hexokinase Pathway
Zhang, W., Adetunji, A. O., Zeng, W., Bastawy, E. M., Min, L., Ding, N., & Zhu, Z. (2025) · Biology · doi:10.3390/biology14091182
“However, sperm exhibit high chromatin aggregation and stable DNA organization, unlike somatic cells [6].”
The Relation of Seminal Cell-Free DNA and Total Antioxidant Capacity with Intracytoplasmic Sperm Injection Outcomes in Iraqi Infertile men
Al-Shammary, H. S., & Al-Omary, H. L. (2025) · F1000Research · doi:10.12688/f1000research.166880.1
Efficiency of sperm separation by using microfluidic chips compared to the swim up method
Ješeta, M., Mekiňová, L., Hošek, J., Ventruba, P., Doubravská, A., & Crha, I. (2025) · Medical Journal of Cell Biology · doi:10.2478/acb-2025-0001
miR-27a rs895819 Polymorphism and Recurrent Pregnancy Loss in Caucasian Women: A Novel Genetic Risk Factor in a Challenging Fertility Dilemma
Panagou, G., Potiris, A., Dedousi, D., Mavrogianni, D., Vassilaki, I., Zikopoulos, A., Moustakli, E., Sfakianakis, A., Kathopoulis, N., Gerede, A., et al. (2025) · Current Issues in Molecular Biology · doi:10.3390/cimb47040271
Comparative analysis of the human sperm cell organelle biochemical markers by confocal Raman spectroscopy
Nazarenko, R., Irzhak, A., Gvasalia, B., & Pushkar, D. (2025) · Bulletin of Russian State Medical University · doi:10.24075/brsmu.2025.016
Maternal Injectable Mineral Administration Effects on Calf Growth and Reproductive Parameters
Brenner, M. A., Marques, R., Posbergh, C., Zezeski, A., Geary, T. W., & Mccoski, S. (2025) · Animals · doi:10.3390/ani15030330
“DNA damage was measured by flow cytometry using acridine orange, as previously reported by Evenson [30].”
Qualitative and quantitative semen parameters in the light of the latest edition of the WHO manual for the examination and processing of human semen
Drlík, M., Krátká, Z., & Novák, J. (2024) · Czech Urology · doi:10.48095/cccu2024015
Effects of Feeding 60% Dried Corn Distillers’ Grains or the Equivalent Sulfur as CaSO4 on DNA Integrity and Gene Expression in Yearling Angus Bull Sperm
Kassetas, C., Geary, T. W., Zezeski, A. L., Caton, J., Kirsch, J. D. D., Dorsam, S., Diniz, W., McCarthy, K., Crouse, M., Sedivec, K., et al. (2023) · Ruminants · doi:10.3390/ruminants3040026
“A DNA compaction assay [25] (IMV Technologies; Maple Grove, MN, USA) was used to estimate the structural stability of sperm chromatin in the nucleus after acid digestion.”
Method
Sperm Chromatin Abnormalities and DNA Damage Predict Assisted Reproductive OutcomePreprint
Bibi, R., Jehan, S., Razak, S., Hammadeh, P. M. E., & Amor, H. (2022) · Research Square · doi:10.21203/rs.3.rs-1231874/v1 · Cited by 1
Study on the Relationship Between Male Infertility and DNA Integrity in Spermatozoa Nuclei
Mitkovska, V., Dzhoglov, S., Boyadzhiev, D., Vasileva, P., Stoyanov, I., Popova, T., Staykova, T., & Ivanova, E. N. (2022) · Proceedings of the Bulgarian Academy of Sciences · doi:10.7546/crabs.2022.01.14
“[11] as follows: excellent – when damaged spermatozoa cells were less than 15%; good – when damaged cells were between 15% and 20%; fair – when damaged cells were between 20% and 30% and poor – when damaged spermatozoa cells were more than 30%.”
Background
The age and mouse sperm quality – a flow cytometry investigationPreprint
Zacchini, F., Bochenek, M., Bisogno, S., Chan, A. B., & Ptak, G. (2021) · bioRxiv (Cold Spring Harbor Laboratory) · doi:10.1101/2021.07.18.451583
Oxidative Stress Testing: Indirect Tests
Sharma, R., Robert, K., & Agarwal, A. (2021) · Manual of Sperm Function Testing in Human Assisted Reproduction · doi:10.1017/9781108878715.018
Use of Novel Methods to Assess Seasonal Differences in the Quality of Boar Semen Stored Up to 7 Days at 17°C
Szczesniak-fabianczyk, B., Bochenek, M., Gogol, P., Trzcińska, M., Bryła, M., & Smorąg, Z. (2019) · Annals of Animal Science · doi:10.2478/aoas-2019-0035
Correlation of sperm DNA integrity with other semen parameters and analyse the role of abnormal sperm DNA in evaluation of infertility
Pushpalatha, D., Tamaskar, S., & Sundaram (2019) · International Journal of Clinical and Diagnostic Pathology · doi:10.33545/pathol.2019.v2.i2g.139
“Similar results were reported by several other studies ([12]).”
Result
Eﬀect of electromagnetic radiation from a mobile phone (1745 MHz) on the condition of the reproductive system of male rats during their postnatal development
Chueshova, N., Vismont, F., & Cheshyk, I. (2019) · Proceedings of the National Academy of Sciences of Belarus Medical series · doi:10.29235/1814-6023-2019-16-2-216-225
Los parámetros espermáticos funcionales como factor pronóstico en ICSI: Experiencia de la ciudad de Medellín
Calderón-Mendoza, L., López, L. C., Giraldo, C. F. V., Álvarez, V. I., & Cardona-Maya, W. (2019) · Revista Urología Colombiana / Colombian Urology Journal · doi:10.1055/s-0039-1688463
Sperm DNA Fragmentation is Important to Predict ART outcomes
Zandiyeh, S. (2018) · Perceptions in Reproductive Medicine · doi:10.31031/prm.2018.03.000551
“The previous studies conducted in humans have demonstrated that it is not always possible to achieve an accurate decision about male fertility according to the results of routine semen examinations [2], and when all the routine sperm parameters are normal, sperm population of an ejaculate may be sub fertile or inferti…”
Background
Metodi di valutazione del danno al DNA dello spermatozoo
Paoli, D., Carlini, T., Faja, F., Muratori, M., & Baldi, E. (2018) · L Endocrinologo · doi:10.1007/s40619-018-00465-1
Image Processing Methods for Automated Assessment of Sperm DNA Integrity
Rekha, B. B., Vidyalakshmi, S., Bose, S., & Devi, T. T. A. (2018) · International Conference on Climate Informatics · doi:10.1007/978-981-13-1132-1_9
Indicative and Prognostic Ramifications in the Assessment of DNA Sperm Damage: The Essential Standards, the Blueprint Procedures, Clinical Procedures, Clinical Importance and Implications of These Tests: A Systematic Review
Margiana, R. (2018) · doi:10.22377/ajp.v12i01.2039
“Sperm chromatin is dealt with in a way thoroughly not the same as that of generous cells.[32,33]”
Background
Outlining adequate protocols for Lidia bull epididymal storage and sperm cryopreservation: use of glycerol, dimethylformamide and N-acetylcysteine
Matilla, E., González-Fernández, L., Martínez-Pastor, F., Hernández, N., Tobajas, C., Calle-Guisado, V., Mijares, J., Sánchez-Margallo, F., Álvarez, I., & Macías-García, B. (2017) · doi:10.5424/sjar/2017153-11463
“Interestingly, DFI below 10% have been related to high fertility status in humans and bulls (Evenson & Wixon, 2006; Waterhouse et al., 2006), suggesting that epididymal Lidia bull sperm fertility is maintained despite prolonged storage.”
Background
La selección espermática mediante técnica de separación magnética de células activadas (MACS) en las técnicas de reproducción asistida
Motato-Moscoso, Y., Ortega-García, A., & Espejo-Catena, M. (2017) · doi:10.1016/j.medre.2017.06.004
“…el potencial de membrana mitocondrial (PMM) y la externalización de la fosfatidilserina (EPS) como resultado de la pérdida de la integridad de la membrana celular (Evenson et al., 2002, Tarozzi et al., 2007), proceso que finalmente permite el reconocimiento por los fagocitos para su eliminación.”
Background
Genetic Testing of Y-Chromosome Microdeletion
Garrido, N., & Rivera, R. (2017) · doi:10.1201/9781315151632-6
The Application of Genetic Tests in an Assisted Reproduction Unit: DNA Fragmentation
Garrido, N., & Rivera, R. (2017) · doi:10.1201/9781315151632-10
Our first experience with the comet assay in the study of sperm DNA integrity
Novotná, B., Svobodova, L., Čechová, M., Jarolím, L., Chocholatý, M., Bagryantseva, Y., & Macek, M. (2016) · Czech Urology · doi:10.48095/cccu2016039
An Evaluation of Equine Sperm Chromatin After Exposure to Ambient Heat Stress
Langdon, W. C., Love, C., Teague, S., Brady, H., & Prien, S. (2015) · doi:10.1016/j.jevs.2015.09.005
Urinary DNA lesions as a biomarker for assessing male reproductive health
Jeng, H., Pan, C.-H., & Chao, M.-R. (2015) · AIMS environmental science · doi:10.3934/environsci.2015.3.565
“[36].”
Etude des dommages de l'ADN impliquant des pontages ADN-protéines et ADN-polyamines
Silerme, S. (2014) · theses.fr (ABES) · doi:10.70675/56da32e9z0503z4bedz8037z6a8e805e6858
Model for cooperation between boar studs and laboratories for reproduction in boars' semen quality control
Milovanović, A., Barna, T., Milanov, D., & Lazarević, M. (2013) · doi:10.46784/e-avm.v6i1.145
Contrôle des dommages oxydants au noyau spermatique : apports des modèles murins knock-out pour des glutathion peroxydases
Noblanc, A. (2013) · doi:10.70675/4e6ce327zdc27z49bfz9d8cz2edf76e4fa8c
Evolution of the Journal of Andrology and a bright future for Andrology.
Meistrich, M., & Carrell, D. (2012) · Journal of Andrology · doi:10.2164/jandrol.112.017368
Relationship between apoptotic markers in semen from fertile men and demographic, hormonal and seminal characteristics.
Specht, I., Spanô, M., Hougaard, K., Manicardi, G., Bizzaro, D., Toft, G., Giwercman, A., & Bonde, J. (2012) · Asian Journal of Andrology · doi:10.1038/aja.2012.76
Utilização da lecitina de soja para a refrigeração e criopreservação do sêmen de cães
Dalmazzo, A., & Barnabé, V. H. (2012) · doi:10.11606/d.10.2012.tde-02102012-105414
Effect of seminal plasma on sperm motility, membrane and DNA integrity in cooled-stored stallion semen
Severgnini, A., Vanni, R., Gaviraghi, A., Bonacina, C., & Galli, A. (2012) · doi:10.1016/j.jevs.2012.06.094
Spermatogenesis Is Not Impaired in a Nucleotide Excision Repair-Deficient Min Mouse Model With or Without Neonatal Mutagen Treatment
Andreassen, Å. K., Steffensen, I., Olsen, A.-K. H., Tanaka, K., & Wiger, R. (2011) · Journal of Andrology · doi:10.2164/jandrol.110.011916
Influência da fragmentação de DNA espermático na produção in vitro de embriões bovinos
Simões, R. (2010) · doi:10.11606/t.10.2010.tde-01042011-145008
Genetic factors of male infertility. The role of complex examination in the case of spermatogenesis disturbances
Brovko, A. A., Тавокина, Л. В., & Sopko, Y. A. (2009) · Cytology and Genetics · doi:10.3103/s0095452709050119
Biochemical markers of male infertility: the key role of DNA damage
Aguilar, C., Garrido, N., Viloria, T., Fernández, J., & Meseguer, M. (2008) · doi:10.1586/17474108.3.4.565
Gamete quality and assisted reproductive technologies
Racowsky, C. (2008) · CRC Press eBooks · doi:10.3109/9780203091500-47
Influence de la congélation sur le taux de fragmentation de l’ADN des spermes normaux à sévèrement altérés
Sabbagh, C., Albert, M., Vialard, F., Hammoud, I., Bergère, M., Delabroye, V., Molina-Gomes, D., Bailly, M., Lombroso, R., & Selval, J. (2007) · doi:10.1007/bf03041156
Instabilité génomique et infertilité masculine
Vialard, F., Benahmed, M., Lombroso, R., & Selva, J. (2004) · Gynécologie Obstétrique & Fertilité · doi:10.1016/j.gyobfe.2004.10.016 · Cited by 2
The use of the acrosome index in assisted reproduction
Menkveld, R. (2004) · doi:10.3109/9780203308547-5
Génotoxicité des chimiothérapies et radiothérapies: Quelles sont les conséquences pour le spermatozoïde humain?
Bujan, L., & Demas, P. (2002) · doi:10.1007/bf03035141
INFLUENCE OF SEMEN QUALITY ON SPERM SURVIVABILITY, SPERM DNA FRAGMENTATION AND FERTILITY RATE OF THE MALE DROMEDARY CAMELS
Zeidan, Ahmadi, E., Amer, A., Mahdy, T. H. M., & Desoky, A. (2020) · doi:10.21608/ejas.2020.112939
“Studies in humans indicate that the frequency of spermatozoa containing fragmented DNA may be a new independent parameter of semen quality and fertility (Evenson et al., 2002).”
Background
ILLUMINATING DNA PACKAGING IN SPERM CHROMATIN: HOW POLYCATION LENGTHS, UNDERPROTAMINATION AND DISULFIDE LINKAGES ALTERS DNA CONDENSATION AND STABILITY
Kirchhoff, D. C. (2019) · doi:10.13023/etd.2019.233
Assessment of Sperm Chromatin Damage by TUNEL Method Using Benchtop Flow Cytometer
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Defective DNA Repair in Spermiogenesis
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Interpreting Semen Analysis and Level 2 Sperm Testing
Aziz, N. (2017) · doi:10.1007/978-3-319-56547-7_2
Evaluation of Semen Oxidative Stress and Sperm DNA Damage in Cases of Unexplained Foetal Congenital Anomalies- A Pilot Study
Kaur, J., Deka, D., Dada, R., Vanamail, P., Dadhwal, V., Malhotra, N., & Kabra, M. (2017) · doi:10.7860/jcdr/2017/24714.10830
Single Gamete Insemination Aiming at the Ideal Conceptus
Neri, Q., Cozzubbo, T., Cheung, S., Rosenwaks, Z., & Palermo, G. (2015) · doi:10.1007/978-3-319-16892-0_6
Evaluation of Men with Unexplained Infertility
Cocuzza, M., & Tiseo, B. (2015) · doi:10.1007/978-1-4939-2140-9_21
Molecular Epidemiology Focused on Airborne Carcinogens
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Assessment and Selection of Human Sperm for ART
Sueldo, C. (2014) · doi:10.1007/978-1-4939-0989-6_3
The Effects of Lead and Cadmium on Sperm Parameters Including Sperm DNA Fragmentation of Men Investigated for Infertility
Wijiesekara, G. D. S. (2013) · doi:10.31357/fmesphd.2013.00123
Estudio de la integridad del ADN espermático en relación con la calidad seminal y los resultados del ciclo de fecundación in vitro
Toledo, M. E. S., Lleó, C. Á., Garrido, C., & de Merlo, G. G. (2013) · doi:10.1016/j.pog.2011.11.008
Import of boars - semen quality control and possibility of complaints.
Milovanović, A., Barna, T., Maksimović, N., Vasiljević, T., Milanov, D., & Bošković, N. (2012) · doi:10.2298/bah1204759m
“…been collected on the results of semen fertility of humans and animals, which indicates that fragmentationdegenaration of sperm DNA has a negative impact on fertility and number of offspring in pluriparous animals (Evenson et al., 2002), where this indicator is not associated with sperm motility.”
BackgroundMethod
Novel Approaches of Sperm Selection for ART: The Role of Objective Biochemical Markers of Nuclear and Cytoplasmic Integrity and Sperm Function
Huszar, G., & Sakkas, D. (2011) · Springer eBooks · doi:10.1007/978-1-4419-8456-2_15
Structure of Chromatin in Human Sperm Bound to Hyaluronic Acid: The Benefits of PICSI Dish Mediated Sperm Selection
Huszar, G., & Sati, L. (2011) · Springer eBooks · doi:10.1007/978-1-4419-6857-9_28
Sperm Chromatin and ART (IUI, IVF and ICSI) Pregnancy
Bungum, M. (2011) · Springer eBooks · doi:10.1007/978-1-4419-6857-9_31
The Clinical Utility of the Evaluation of Sperm Chromatin
Moskovtsev, S., & Mullen, B. (2010) · doi:10.1007/978-1-4419-1436-1_30
Sperm Chromatin Abnormalities and Reproductive Outcome
Schlegel, P. (2009) · doi:10.1007/978-1-60327-392-3_9

Authors: EVENSON, D. P., LARSON, K. L., & JOST, L. K.
Affiliations: South Dakota State University (US)
Corresponding author: EVENSON, DONALD P.
ORCID: EVENSON, DONALD P.
Venue: Journal of Andrology
Publisher: Wiley
Published: 2002-01-02
Volume: 23
Issue: 1
Pages: 25-43
ISSN: 0196-3635 (ISSN-L)
1939-4640
DOI: 10.1002/j.1939-4640.2002.tb02599.x
OpenAlex: W1532901402
Cited by: 1,072
References: 158
Citations by year: 2026: 15 · 2025: 21 · 2024: 18 · 2023: 25 · 2022: 36 · 2021: 59 · 2020: 35 · 2019: 50 · 2018: 49 · 2017: 51 · 2016: 42 · 2015: 53 · 2014: 53 · 2013: 68 · 2012: 61
FWCI: 38.26
Type: journal article
Language: en
Open access: bronze
doi.org
Licence: onlinelibrary.wiley.com/termsAndConditions#vor (vor, 4018 d)
Locations: Journal of Andrology (publishedVersion)
PubMed (publishedVersion)
Funders: National Science Foundation (9452894, 9720642)
U.S. Environmental Protection Agency
South Dakota Agricultural Experiment Station
Topics: Health Sciences › Medicine › Reproductive Medicine › Sperm and Testicular Function
Life Sciences › Biochemistry, Genetics and Molecular Biology › Physiology › Reproductive biology and impacts on aquatic species
Health Sciences › Medicine › Urology › Male Reproductive Health Studies
Keywords: sperm chromatin structure assay, sperm DNA fragmentation, male infertility, testicular cancer
MeSH terms: Chromatin, Humans, Infertility, Male, Male, Spermatozoa, DNA Fragmentation

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Sperm Chromatin Structure Assay: Its Clinical Use for Detecting Sperm DNA Fragmentation in Male Infertility and Comparisons With Other Techniques

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