Journal of Medical Diagnostic Methods
Open Access

Department of Obstetrics and Gynaecology, Ichikawa General Hospital, Tokyo Dental College, Ichikawa, Japan

Publications

• Research Article   
  Revalidation of the Sperm Chromatin Dispersion Test and the Comet Assay Using Intercomparative Studies Between Purified Human Sperm without and with End-Stage DNA Fragmentation
  Author(s): Satoru Kaneko* and Kiyoshi Takamatsu
  
  Objectives: To perform intercomparative studies to revalidate the principles and quantitative performance of the Sperm Chromatin Dispersion Test (SCD) and Comet Assay (CA). Methods: Human sperm without and with granular fragments in end-stage fragmentation were purified from normozoospermic semen and used as naturally occurring negative and positive controls (NC and PC, respectively). Both SCD and CA extracted nucleoproteins with 2.0 mol/L NaCl, 1.0 mmol/L DTT. SCD determined that DNA damage as inversely proportional to the area of the violet halo. The CA estimated the level of DNA damage electrophoretically from the number of granular fragments; the so-called comet tail, their electrophoretic features were compared with the Single-Cell Pulsed-Field Gel Electrophoresis (SCPFGE). Results: The violet ha.. View More»
  DOI: 10.35248/2168- 9784.23.12.406

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