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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“Así, la determinación del estado del ADN se configura como un parámetro independiente en el estudio de la calidad seminal y un potencial instrumento en la predicción de la fertilidad (Evenson, 2002).”
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“An aliquot of the whole semen was diluted in tris NaCl EDTA (TNE) buffer with glycerol and frozen for the SCSA analysis (Evenson et al., 2002).”
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“Sperm DNA Fragmentation Index (DFI) and high DNA stainability (HDS) were measured by the sperm chromatin structure (SCSA) technique (Evenson et al., 2002) routinely used in our laboratory (Pecou et al., 2009; Ahmad et al., 2012).”
MethodInfluential
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“However, abnormal chromatin structure can increase the susceptibility to in situ acid-induced DNA denaturation, and the Sperm Chromatin Structure Assay (SCSA) (Evenson et al. 2002) measures the proportion of sperm being denatured after exposure to low pH.”
Activation of the immune system and sperm DNA fragmentation are associated with idiopathic oligoasthenoteratospermia in men with couple subfertility.
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“Other less commonly used techniques involve detecting defective sperm organelles and DNA [10,11] and the analysis of seminal plasma [12,13].”
Method
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“The stability of sperm chromatin construction was assessed through the utilization of the sperm chromatin structure test, following previous methods [60, 61] employing an acridine orange (AO) fluorescence staining kit (Genmed Scientifics Inc., HL14018.”
Method
Effects of Chokeberries (Aronia spp.) on Cytoprotective and Cardiometabolic Markers and Semen Quality in 109 Mildly Hypercholesterolemic Danish Men: A Prospective, Double-Blinded, Randomized, Crossover Trial
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“This test is based on the staining protocol for the SCSA® [58] but with a more detailed quality control in the flow.”
Method
Telomere Distribution in Human Sperm Heads and Its Relation to Sperm Nuclear Morphology: A New Marker for Male Factor Infertility?
Turner, K. J., Watson, E. M., Skinner, B. M., & Griffin, D. K. (2021) · International Journal of Molecular Sciences · doi:10.3390/ijms22147599 · Cited by 12
DNA Fragmentation in Human Spermatozoa and Pregnancy Rates after Intrauterine Insemination. Should the DFI Threshold Be Lowered?
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“The flow cytometric assay is believed to be the most objective and robust way to analyse DNA fragmentation as it, among other advantages, provides an objective analysis of thousands of cells in a short period of time [3,4].”
Background
Compound heterozygosity for novel AURKC mutations in an infertile man with macrozoospermia
Bai, S., Hu, X., Zhao, Y., Li, W., Wan, Y., Jin, R.-T., Wang, Y.-S., Guo, T., Tong, X., & Xu, B. (2020) · Andrologia · doi:10.1111/and.13663 · Cited by 11
DNA Fragmentation in Viable and Non-Viable Spermatozoa Discriminates Fertile and Subfertile Subjects with Similar Accuracy
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“[37] to explain why subjects with sDF over a threshold of 30% resulted infertile despite the remaining 70% of spermatozoa, apparently without DNA damage.”
Background
Ovine sperm DNA oxidation quantification using an 8-OHdG immunodetection assay.
Soria-Meneses, P. J., Jurado-Campos, A., Montoro, V., Soler, A., Garde, J., & Fernández-Santos, M. D. R. (2019) · Reproduction in domestic animals = Zuchthygiene · doi:10.1111/rda.13533 · Cited by 11
Sperm chromatin alterations in fertile and subfertile bulls.
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Improved methods of DNA extraction from human spermatozoa that mitigate experimentally-induced oxidative DNA damage
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Induced sub-lethal oxidative damage affects osmotic tolerance and cryosurvival of spermatozoa.
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“Flow cytometric analysis of chromatin stability The sperm chromatin structure assay (SCSA) was used to evaluate chromatin integrity (Evenson et al. 2002).”
Method
Mitochondrial DNA Copy Number in Spermatozoa of Fertile Stallions.
Orsztynowicz, M., Pawlak, P., Podstawski, Z., Niżański, W., Partyka, A., Gotowiecka, M., Kosiniak-Kamysz, K., & Lechniak, D. (2016) · Reproduction in domestic animals = Zuchthygiene · doi:10.1111/rda.12689 · Cited by 11
DNA fragmentation of human sperm can be detected by ligation-mediated real-time polymerase chain reaction.
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Semen quality assessments and their significance in reproductive technology.
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“The SCSA predicts infertility when the DFI exceeds 30% in natural conception (Evenson et al. 2002, Evenson and Wixon 2008), justifying its clinical application.”
Background
Sperm DNA Fragmentation Assays Correlate with Sperm Abnormal Morphology and the Pregnancy Outcome
Fortunato, A., Leo, R., Casale, S., Nacchia, G., Liguori, F., & Tosti, E. (2013) · doi:10.4172/2375-4508.1000101 · Cited by 11
“Although a recognized threshold value for DFI has not been established yet, in this work we referred to some recent studies that have indicated: i) 30% as the cut off value above that natural pregnancy is not possible to be achieved [3,24] ii) DFI > 30% provides significantly lower embryo implantation rate and pregnan…”
BackgroundResult
Causes et implications cliniques des altérations de l’ADN des spermatozoïdes
Hazout, A., Ménézo, Y., Madelenat, P., Yazbeck, C., Selva, J., & Cohen-Bacrie, P. (2008) · doi:10.1016/j.gyobfe.2008.07.017 · Cited by 11
Effect of repeated sequential ejaculation on sperm DNA integrity in subfertile males with asthenozoospermia
Hussein, T., Elariny, A. F., Elabd, M., Elgarem, Y. F., & Elsawy, M. (2008) · Andrologia · doi:10.1111/j.1439-0272.2008.00861.x · Cited by 11
“The clinical significance of this assessment lies in its association not only with natural conception rates, but also with assisted reproduction success rates (Evenson et al., 2002).”
Background
CONSERVATION SCIENCE IN A TERRORIST AGE: THE IMPACT OF AIRPORT SECURITY SCREENING ON THE VIABILITY AND DNA INTEGRITY OF FROZEN FELID SPERMATOZOA
Gloor, K., Winget, D., & Swanson, W. (2006) · Journal of zoo and wildlife medicine · doi:10.1638/05-072.1 · Cited by 11
Lonidamine transiently affects spermatogenesis in pubertal CD1 mice.
Traina, M. E., Guarino, M., Urbani, E., Saso, L., Eleuteri, P., Cordelli, E., Rescia, M., Leter, G., & Spanô, M. (2005) · Contraception · doi:10.1016/j.contraception.2005.04.008 · Cited by 11
“Adopting the guidelines described in a recent publication [24] and using the dedicated SCSASoft software (SCSA Diagnostic, Brookings, SD, USA) for offline data analysis, we have expressed the extent of DNA denaturation in terms of a DNA fragmentation index (DFI), which is the ratio of red to total (red+green) fluoresc…”
Method
The higher risk for sperm DNA damage in infertile men.
Gill, K., Rosiak-Gill, A., Jakubik, J., Patorski, L., Lukaszuk, M., & Piasecka, M. (2019) · Ginekologia Polska · doi:10.5603/gp.2019.0117 · Cited by 11
Oxidative Stress, Sperm DNA Fragmentation, or Both? Optimizing Test Selection in Male Infertility Evaluation
Kaltsas, A., Papaharitou, S., Sengupta, P., Saleh, R., & Agarwal, A. (2026) · Antioxidants · doi:10.3390/antiox15030293 · Cited by 11
“…harmonized protocols and population-based reference ranges are still lacking for key assays, particularly TUNEL, Comet, chemilumines-cent ROS, TAC/FRAP, and ORP, so laboratories therefore rely on internally derived cut-offs that complicate cross-center comparison and meta-analysis [148,161–163].”
Impact of sperm DNA fragmentation in couples with unexplained recurrent pregnancy loss: A cross‐sectional study
Solanki, M., Joseph, T., Muthukumar, K., Samuel, P., Aleyamma, T. K., & Kamath, M. S. (2024) · Journal of obstetrics and gynaecology research · doi:10.1111/jog.16041 · Cited by 9
“Secondary outcomes included the proportion of men with high and low DFI (as defined as a cut-off DFI ≥ 30% 14,25 ) in the unexplained RPL versus control groups.”
Determination of Ram (Ovis aries) Sperm DNA Damage Due to Oxidative Stress: 8-OHdG Immunodetection Assay vs. SCSA®
Soria-Meneses, P. J., Jurado-Campos, A., Gómez-Rubio, V., Sánchez-Ajofrín, I., Soler, A., Garde, J., & Fernández-Santos, M. D. R. (2022) · Animals · doi:10.3390/ani12233286 · Cited by 10
The relationship between reactive oxygen species, DNA fragmentation, and sperm parameters in human sperm using simplified sucrose vitrification with or without triple antioxidant supplementation
Juanpanich, T., Suttirojpattana, T., Parnpai, R., & Vutyavanich, T. (2022) · Clinical and Experimental Reproductive Medicine · doi:10.5653/cerm.2021.05120 · Cited by 10
“HDS is considered as indicating immature spermatozoa [24].”
BackgroundMethodInfluential
Drying and temperature induced conformational changes of nucleic acids and stallion sperm chromatin in trehalose preservation formulations
Brogna, R., Fan, J., Sieme, H., Wolkers, W. F., & Oldenhof, H. (2021) · Scientific Reports · doi:10.1038/s41598-021-93569-y · Cited by 10
Importance of a semen analysis report for determining the relationship between SCSA sperm DNA fragmentation index and assisted reproductive technology pregnancy rate.
Cheng, H.-B., Han, M.-T., Ding, J., Wang, F.-X., Wang, G.-G., Shen, L.-Y., Wang, J.-X., Zheng, B., Meng, Q., Wang, W., et al. (2020) · Reproductive Biology · doi:10.1016/j.repbio.2019.09.002 · Cited by 10
Effects of cashew gum and nanoparticles on cooled stallion semen
Loureiro, K., Lima-Verde, I., Johannisson, A., Ntallaris, T., Jager, A., Štěpánek, P., Mendonça, M. D. C., Severino, P., & Morrell, J. (2020) · Acta Veterinaria Scandinavica · doi:10.1186/s13028-020-00530-6 · Cited by 10
Comparative Cytotoxicity and Genotoxicity Assessments of Chitosan Amino Acid Derivative Nanoparticles Toward Human Breast Cancer Cell Lines
Zaboon, M., Saleh, A., & Al-Lami, H. (2019) · Egyptian Journal of Chemistry · doi:10.21608/ejchem.2019.10806.1708 · Cited by 10
SCSA results correlated with rate of motility reduction after ejaculation in Asthenozoospermia
Fard, Z. M., Naghdi, M., Salehi, P., Shahrokhi, S. Z., Ajami, A., Deemeh, M., & Meshkibaf, M. (2018) · Andrologia · doi:10.1111/and.13146 · Cited by 10
In vitro studies of Norwegian Red bovine semen immobilized and cryopreserved in alginate solid gel network
Alm-Kristiansen, A., Gaustad, E., Bai, G., Standerholen, F., Klinkenberg, G., Kommisrud, E., & Waterhouse, K. (2018) · Reproduction in domestic animals = Zuchthygiene · doi:10.1111/rda.13115 · Cited by 10
“The SCSA® measures the susceptibility of sperm nuclear DNA to acid- induced denaturation in situ (Evenson et al., 2002) and is inversely related to chromatin organization and DNA strand breaks (Sailer, Jost, & Evenson, 1995; Waterhouse et al., 2006).”
BackgroundMethodInfluential
Cross Breeding Promotes Deterioration of Semen Quality in Cattle Bulls
Khan, I., Khan, R., Qureshi, M. S., Usman, T., Khan, A., Ullah, Z., & Rehman, H. (2017) · doi:10.17582/journal.pjz/2018.50.1.97.103 · Cited by 10
The Effect of Oxidative Stress on Thawed Bulk-Sorted Red Deer Sperm.
Anel-López, L., García-Álvarez, O., Parrilla, I., Olmo, D. D. D., Fernández-Santos, Soler, A., Maroto-Morales, A., Ortiz, J., Alkmin, D., Tarantini, T., et al. (2016) · Reproduction in domestic animals = Zuchthygiene · doi:10.1111/rda.12694 · Cited by 10
“Chromatin stability was assessed following the SCSA (Sperm Chromatin Structure Assay), based on the susceptibility of sperm DNA to acid-induced denaturation in situ and on the subsequent staining with the metachromatic fluorescent dye acridine orange (Evenson et al. 2002).”
Method
Effect of cryopreservation on the sperm DNA fragmentation dynamics of the bottlenose dolphin (Tursiops truncatus).
Sánchez-Calabuig, M., López-Fernández, C., Johnston, S. D., Blyde, D., Cooper, J., Harrison, K., de la Fuente, J. M., & Gosálvez, J. (2015) · Reproduction in domestic animals = Zuchthygiene · doi:10.1111/rda.12474 · Cited by 10
“A number of techniques for assessing SDF have been developed including the comet assay (Hughes et al. 1996; Aravindan et al. 1997; Singh et al. 1998; Gwo et al. 2003; Fraser and Strzezek 2004), the TUNEL assay (terminal deoxynucleotidyl transferase-mediated ISSN 0936–6768 © 2015 Blackwell Verlag GmbH dUTP nick end lab…”
BackgroundMethod
Characteristics of high-quality Asian elephant (Elephas maximus) ejaculates and in vitro sperm quality after prolonged chilled storage and directional freezing.
O'Brien, J., Steinman, K., Montano, G., Love, C., Saiers, R. L., & Robeck, T. (2013) · Reproduction, Fertility and Development · doi:10.1071/rd12129 · Cited by 10
“For evaluation of sperm DNA integrity, a sample aliquot (55 mL) was snap frozen in liquid nitrogen at each step and retrospectively analysed for the susceptibility of spermatozoa to DNA denaturation using the sperm chromatin structure assay (SCSA; Evenson et al. 2002).”
Method
Assessment of buffalo (Bubalus bubalis) sperm DNA fragmentation using a sperm chromatin dispersion test.
Pawar, K., & Kaul, G. (2011) · Reproduction in domestic animals = Zuchthygiene · doi:10.1111/j.1439-0531.2011.01766.x · Cited by 10
APEX/Ref-1 (apurinic/apyrimidic endonuclease DNA-repair gene) expression in human and ascidian (Ciona intestinalis) gametes and embryos.
El-Mouatassim, S., Bilotto, S., Russo, G., Tosti, E., & Ménézo, Y. (2007) · Molecular human reproduction · doi:10.1093/molehr/gam038 · Cited by 10
“More recently, careful attention has been paid to the quality of sperm DNA, and new methods have been developed to evaluate paternal DNA integrity, which can be totally independent of all semen parameters, including sperm morphology, concentration and motility (Evenson et al., 2002; Oger et al., 2003).”
[Gamete and embryo protection against oxidative stress during medically assisted reproduction].
Ménézo, Y., & Guérin, P. (2005) · Bulletin de l'Académie Nationale de Médecine · doi:10.1016/s0001-4079(19)33547-2 · Cited by 10
Epigallocatechin‐3‐gallate (EGCG) and green tea polyphenols do not improve stallion semen parameters during cooling at 4°C
Bucci, D., Spinaci, M., Mislei, B., Gadani, B., Rizzato, G., Love, C., Tamanini, C., Galeati, G., & Mari, G. (2017) · Reproduction in domestic animals = Zuchthygiene · doi:10.1111/rda.12891 · Cited by 10
Reproductive and Developmental Toxicity of Metals
Apostoli, P., Telišman, S., & Sager, P. R. (2007) · Elsevier eBooks · doi:10.1016/b978-012369413-3/50067-7 · Cited by 27
Distinguishing between fertilization failure and early pregnancy loss when identifying male-mediated adverse pregnancy outcomes.
Perreault, S. (2003) · Advances in Experimental Medicine and Biology · doi:10.1007/978-1-4419-9190-4_16 · Cited by 10
Sperm Quality in Young Bull Semen Can Be Improved by Single Layer Centrifugation
Lima-Verde, I., Hurri, E., Ntallaris, T., Johannisson, A., Stålhammar, H., & Morrell, J. (2022) · Animals · doi:10.3390/ani12182435 · Cited by 9
“The method is a modification of [18] as reported by [19].”
Method
Expression of membrane fusion proteins in spermatozoa and total fertilisation failure during in vitro fertilisation
Enoiu, S. I., Nygaard, M. B., Bungum, M., Ziebe, S., Petersen, M. R., & Almstrup, K. (2022) · Andrology · doi:10.1111/andr.13215 · Cited by 11
Advanced sperm testing
Softness, K. A., Trussler, J. T., & Carrasquillo, R. J. (2020) · Current Opinion in Urology · doi:10.1097/mou.0000000000000761 · Cited by 13
Sperm DNA fragmentation: overcoming standardization obstacles
Majzoub, A., Agarwal, A., & Esteves, S. (2017) · Translational Andrology and Urology · doi:10.21037/tau.2017.07.10 · Cited by 9
A data‐driven search for semen‐related phenotypes in conception delay
Patel, C. J., Sundaram, R., & Louis, G. M. B. (2016) · Andrology · doi:10.1111/andr.12288 · Cited by 12
“SCSA was measured on the first specimen (Evenson et al., 2002) and using a Coulter Epics Elite Flow Cytometer and software (SCSA diagnostics, Brookings, SD, USA).”
Method
The Importance of Semen Quality in AI Programs and Advances in Laboratory Analyses for Semen Characteristics Assessment
Oliveira, L. Z., Monteiro, F. M., de Arruda, R. D., & Celeghini, E. C. (2013) · doi:10.5772/52022 · Cited by 9
Combination of swim-up and density gradient separation methods effectively eliminate DNA damaged sperm.
Ghumman, S., Adiga, S., Upadhya, D., Kalthur, G., Jayaraman, V., Rao, S., & Kumar, P. (2011) · Journal of the Turkish-German Gynecological Association · doi:10.5152/jtgga.2011.35 · Cited by 9
“There has been increased concern regarding the role of sperm DNA integrity in male infertility (1, 2).”
Background
Modern Andrology
Patrizio, P., Sanguineti, F., & Sakkas, D. (2008) · Annals of the New York Academy of Sciences · doi:10.1196/annals.1434.021 · Cited by 9
Chapter 6 – Semen Evaluation
Sieme, H. (2009) · doi:10.1016/b978-1-4160-5234-0.00006-4 · Cited by 9
Zishen Yutai Pill improves sperm quality and reduces testicular inflammation in experimental varicocele rats
Peng, M.-L., Wang, W., Zhu, W., Bai, Y., Ning, N., Huang, Q.-L., Pang, X., Zhou, J., Zhang, H.-P., & Zhao, K. (2023) · Heliyon · doi:10.1016/j.heliyon.2023.e17161 · Cited by 8
“Epididymal sperm DNA fragmentation index (DFI) was analyzed using SCSA (sperm chromatin structure assay) as previously described [26].”
Method
Molecular analysis of microorganisms in the semen and their impact on semen parameters.
Suárez, J. P., Hernández, J. C., & Maya, W. C. C. (2022) · Archivio Italiano di Urologia e Andrologia · doi:10.4081/aiua.2022.2.199 · Cited by 8
“Sperm mitochondrial membrane potential (24), sperm membrane integrity (25), chromatin structure assay (26), sperm membrane lipoperoxidation (27), and intracellular levels of reactive oxygen species (ROS) (24) were evaluated by ﬂow cytometry ( Fortessa-Becton Dickinson, NJ, USA ), according to…”
The Effect of Sperm DNA Fragmentation on In Vitro Fertilization Outcomes for Women With Polycystic Ovary Syndrome
Wang, H., Li, H., Zhu, J., Xu, J., Jiang, Y., Chen, W., Sun, Y., & Yang, Q. (2022) · Frontiers in Endocrinology · doi:10.3389/fendo.2022.822786 · Cited by 7
“However, the SCSA is different from other tests, which is carried out under a normalized protocol (31) and has definite and clinically valuable cutoff levels for evaluating spermatogenic potential (32).”
Background
Post-thaw semen quality in young bull ejaculates before being accepted for commercial semen doses.
Hurri, E., Lima-Verde, I., Johannisson, A., Stålhammar, H., Ntallaris, T., & Morrell, J. (2022) · The Veterinary Record · doi:10.1002/vetr.1386 · Cited by 8
Sperm deoxyribonucleic acid fragmentation: predictors, fertility outcomes, and assays among infertile males
Rios, J., Coward, R., Hansen, K., Barnhart, K., Cedars, M., Legro, R., Diamond, M., Krawetz, S., Usadi, R., Baker, V., et al. (2021) · F&S Reports · doi:10.1016/j.xfre.2021.06.003 · Cited by 8
“analysis was performed, and 3 aliquots of neat semen were stored at 80 C and shipped to the Utah Andrology Laboratory for DNA fragmentation analysis by SCSA (19), TUNEL, and Comet assays (20).”
Season does not have a deleterious effect on proportions of stallion seminal plasma proteins
Johannisson, A., Al-Essawe, E., Al-Saffar, A., Karkehabadi, S., Lima-Verde, I., Wulf, M., Aurich, C., & Morrell, J. (2020) · Journal of reproduction and development · doi:10.1262/jrd.2019-072 · Cited by 8
Assessing the potential of HSPA2 and ADAM2 as two biomarkers for human sperm selection
Heidari, M., Darbani, S., Darbandi, M., Lakpour, N., Fathi, Z., Zarnani, A., Zeraati, H., Akhondi, M., & Sadeghi, M. (2018) · Human Fertility · doi:10.1080/14647273.2018.1534277 · Cited by 8
“The assessment of DFI was carried out using the sperm chromatin structure assay (SCSA) method according to Evenson et al. (2002).”
Method
Association of hsa-miR-145 overexpression in human testicular cells with male infertility
Zhang, L., Ding, X., Nie, S., Li‐Ling, J., Zhang, Y., Zhang, H., Chen, L., Li, L., & Ding, M. (2015) · Molecular Medicine Reports · doi:10.3892/mmr.2015.3273 · Cited by 8
“In at least one-third of male infertility cases, the cause is elusive (2) and is referred to as idiopathic infertility.”
Localisation and quantification of alkali-labile sites in human spermatozoa by DNA breakage detection-fluorescencein situhybridisation
Cortés-Gutiérrez, E. I., Dávila-Rodríguez, M. I., Cerda-Flores, R. M., Fernández, J. L., López‐Fernández, C., Tovar, A. R. A., & Gosálvez, J. (2014) · Andrologia · doi:10.1111/and.12250 · Cited by 12
“DNA damage in the male germ line is associated with fertilisation failure, impaired pre- and post-implantation development and poor pregnancy outcomes (Evenson et al., 2002).”
Background
Validation of a field based chromatin dispersion assay to assess sperm DNA fragmentation in the bottlenose dolphin (Tursiops truncatus).
Sánchez-Calabuig, M., López-Fernández, C., Martínez-Nevado, E., Perez-Gutierrez, J., de la Fuente, J. M., Johnston, S. D., Blyde, D., Harrison, K., & Gosalvez, J. (2014) · Reproduction in domestic animals = Zuchthygiene · doi:10.1111/rda.12364 · Cited by 8
“The assessment of DNA fragmentation represents an important tool for the prediction of male infertility (Evenson et al. 2002; Agarwal and Allamaneni 2005).”
BackgroundMethod
Evaluation of sperm functional parameters in normozoospermic infertile individuals
Mayorga-Torres, B. J. M., Cardona-Maya, W., Cadavid, Á., & Camargo, M. (2013) · doi:10.1016/j.acuroe.2012.06.007 · Cited by 8
Significance of extruded nuclear chromatin (regional nuclear shape malformation) in human spermatozoa: implications for ICSI.
Mauri, A., Oliveira, J. B., Baruffi, R., Petersen, C., Vagnini, L., Massaro, F., Silva, L. F. I., Nicoletti, A., & Franco, J. (2011) · International Journal of Andrology · doi:10.1111/j.1365-2605.2010.01119.x · Cited by 8
“…negatively with rates of fertilization, embryonic cleavage, implantation and live birth and positively with miscarriage rates (Larson et al., 2000; Evenson et al., 2002; Morris et al., 2002; Benchaib et al., 2003; Carrell et al., 2003; Saleh et al., 2003; Agarwal & Allameneni, 2004; Evenson &…”
Factors affecting fecundity among sperm donors: a multivariate analysis
Hu, J., Zhu, W.-B., Liu, W., & Fan, L.-Q. (2011) · Andrologia · doi:10.1111/j.1439-0272.2009.01036.x · Cited by 8
“Sperm chromatin structure assay (SCSA) The SCSA protocol has been described by Evenson and Jost (Evenson et al., 1999, 2002).”
Method
Detection of DNA damage in sperm
Henkel, R. (2007) · doi:10.3109/9780203090626-24 · Cited by 8
Sperm Chromatin Structure Assay (SCSA®): Evolution from Origin to Clinical Utility
Evenson, D. (2018) · doi:10.1007/978-3-319-71815-6_4 · Cited by 8
Self-citation
Oxidative Stress in Unexplained Male Infertility
Doshi, S., Sharma, R., & Agarwal, A. (2015) · doi:10.1007/978-1-4939-2140-9_10 · Cited by 8
Understanding the Spermatozoon
Neri, Q. V., Hu, J., Rosenwaks, Z., & Palermo, G. D. (2014) · Methods in molecular biology · doi:10.1007/978-1-4939-0659-8_5 · Cited by 9
A machine learning approach using semen parameters and sperm mitochondrial DNA copy number to predict couples’ fecundity
Sawant, S., Paskavitz, A. L., Nowak, K., Rosati, A., Westling, T., Bertolla, R. P., Whitcomb, B. W., & Pilsner, J. R. (2025) · F&S Reports · doi:10.1016/j.xfre.2025.05.002 · Cited by 4
“DNA integrity parameters, such as DNA fragmentation index (DFI) and high DNA stain-ability (HDS), were measured using the sperm chromatin structural assay (5, 20).”
Semen quality parameters including metabolites, sperm production traits and fertility in young Norwegian Red AI bulls
Narud, B., Khezri, A., Nordborg, A., Klinkenberg, G., Zeremichael, T. T., Stenseth, E.-B., Heringstad, B., Kommisrud, E., & Myromslien, F. D. (2021) · Livestock Science · doi:10.1016/j.livsci.2021.104803 · Cited by 7
Sperm chromatin dispersion test (SCDt) for the assessment of sperm DNA fragmentation in black tiger prawn, Penaeus monodon
Feng, T., Gosálvez, J., López-Fernández, C., Arroyo, F., Paterson, B., & Johnston, S. (2018) · doi:10.1016/j.aquaculture.2018.03.041 · Cited by 7
A pilot study on sperm DNA damage in β-thalassemia major: is there a role for antioxidants?
Elsedfy, H., Sanctis, V. D., Ahmed, A. T., Mohamed, N. R., Arafa, M., & Elalfy, M. S. (2018) · Acta bio-medica : Atenei Parmensis · doi:10.23750/abm.v89i1.6836 · Cited by 7
“Increased sperm DNA FI was consistently associated with reduced fertility in several studies (32-34).”
Background
Aneuploidy frequency in spermatozoa of Egyptian men with normal and abnormal semen parameters using fluorescence in situ hybridisation
Younan, D., Sorour, A., & Genedy, R. (2015) · Andrologia · doi:10.1111/and.12251 · Cited by 7
“Accumulating evidence shows that aneuploidies could be indicative of male subfertility regardless of sperm concentration, per cent motility and morphology (Evenson et al., 2002).”
Background
Evaluation of chromatin integrity of motile bovine spermatozoa capacitated in vitro
Rečková, Z., Machatkova, M., Rybář, R., Horakova, J., Hulínská, P., & Máchal, L. (2008) · Zygote (Cambridge. Print) · doi:10.1017/s0967199408004772 · Cited by 7
“3Department of Animal Breeding, Mendel University of Farming and Forestry, Zemedelska 1, 613 00 Brno, Czech Republic. been described by Evenson et al. (2002).”
Background
[Indications for IMSI].
Cohen-Bacrie, P., Dumont, M., Junca, A., Belloc, S., & Hazout, A. (2007) · Journal de gynecologie, obstetrique et biologie de la reproduction · doi:10.1016/s0368-2315(07)78741-0 · Cited by 7
Apoptosis in Ejaculated Spermatozoa and in the Normal and Pathological Testes: Abortive Apoptosis and Sperm Chromatin Damage
Sakkas, D., & El-Fakahany, H. (2018) · doi:10.1007/978-3-319-71815-6_12 · Cited by 7
Sperm DNA damage causes, assessment and relationship with fertility: A review
Lone, S., Shah, N., Yadav, H., Wagay, M. A., Singh, A., & Sinha, R. (2017) · doi:10.5958/2277-3371.2017.00010.9 · Cited by 7
“well establish differences between chromatin packaging density between humans and rams, the %DFI in rams is much lower (Evenson et al., 2002).”
Background
Sperm Quality and Function Tests
Yeung, C., & Cooper, T. (2010) · doi:10.1007/978-3-540-78355-8_10 · Cited by 7
“In the latter assay, the ratio of red (R) and green (G) [R/R+G] provides the DNA fragmentation index (DI) refl ecting DNA strand breaks (Evenson et al. 2002).”
BackgroundInfluential
Impact Of Air Pollution On Reproductive Health In Northern Bohemia
Rubes, J., Selevan, S., Sram, R., Evenson, D., & Perreault, S. (2007) · doi:10.1007/1-4020-4829-7_10 · Cited by 7
Self-citation
Effects of long-term cryopreservation on sperm quality and oxidative stress in Manchega rams (Ovis aries)
Soria-Meneses, P. J., Jurado-Campos, A., de Juan, A. R., Angulo, V. M., Soler, A. J., Garde, J., Fernández, M. R., & Fernández-Santos, M. D. R. (2025) · Reproduction · doi:10.1530/rep-24-0402 · Cited by 6
“Flow cytometry data were processed to obtain %DFI (% of spermatozoa with DFI >25) and %HDS, which is the percentage of spermatozoa with green ﬂ uorescence higher than channel 600 of 1,024 channels (Evenson et al. 2002).”
High sperm DNA stainability might not be an accurate predictive indicator of male fertility and assisted reproductive technology outcomes
Shen, L.-Y., Zhang, C., Wang, G.-G., Fu, X., Yang, S., & Wang, J. (2025) · Frontiers in Endocrinology · doi:10.3389/fendo.2025.1510114 · Cited by 6
“This assay is based on the characteristics of Acridine Orange (AO), which is a cell-permeable dye that shows ﬂ uoresces green when bound to native double-stranded DNA and yellow/red when bound to single-stranded DNA (12).”
The impact of female BMI on sperm DNA damage repair ability of oocytes and early embryonic development potential in intracytoplasmic sperm injection cycles
Jiang, Y., Shen, Z., Xu, J., Zhu, J., Wang, H., Chen, W., Sun, Y., & Yang, Q.-L. (2023) · Frontiers in Endocrinology · doi:10.3389/fendo.2023.1168010 · Cited by 6
“Even if a sperm with normal motility and morphology is selected and injected directly into the cytoplasm of amature oocyte during the ICSI cycle, the selected spermatozoa may have a defective genome with a high DFI (41).”
Sperm competition increases sperm production and quality in Cataglyphis desert ants
Degueldre, F., & Aron, S. (2023) · Proceedings of the Royal Society B · doi:10.1098/rspb.2023.0216 · Cited by 6
The Quality and Fertilizing Potential of Red Deer (Cervus elaphus L.) Epididymal Spermatozoa Stored in a Liquid State
Dziekońska, A., Koziorowska-Gilun, M., Kordan, W., Neuman, N. M., Kotlarczyk, A. M., & Korzekwa, A. (2022) · International Journal of Molecular Sciences · doi:10.3390/ijms232314591 · Cited by 6

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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