The Sperm Chromatin Structure Assay (SCSA(®)) and other sperm DNA fragmentation tests for evaluation of sperm nuclear DNA integrity as related to fertility.

摘要: Thirty-five years ago the pioneering paper in Science (240:1131) on relationship between sperm DNA integrity and pregnancy outcome was featured as cover issue showing a fluorescence photomicrograph of red green stained sperm. The flow cytometry data showed very significant difference fertile subfertile bulls men. This study utilized heat (100°C, 5min) to denature at sites strand breaks followed by staining with acridine orange (AO) measurements 5000 individual double (ds) single (ss) fluorescence. Later, protocol changed low pH breaks; acid procedures produced same results. SCSA are advantageously dual parameter 1024 channels (degrees) both Hundreds publications use test animals humans have validated highly useful for determining male breeding soundness. is rapid, non-biased cytometer machine measurement providing robust statistical exceptional precision repeatability. Many genotoxic experiments excellent dose response coefficient variation that further being powerful assay integrity. Twelve following introduction test, terminal deoxynucleotidyl transferase-mediated fluorescein-dUTP nick end labelling (TUNEL) (1993) introduced only other cytometric fragmentation. However, TUNEL can also be done light microscopy much less robustness. COMET (1998) Sperm Chromatin Dispersion (SCD; HALO) (2003) tests were microscope don't require cytometer. Since these measure 50-200 per sample, they suffer from lack robustness measurements. Only has an exact standardization fixed protocol. many variations make it difficult compare thresholds risk factor infertility. Data four fragmentation plus (AOT) correlated various degrees.