PMID- 22992911
OWN - NLM
STAT- MEDLINE
DCOM- 20130124
LR  - 20220410
IS  - 1940-6029 (Electronic)
IS  - 1064-3745 (Linking)
VI  - 927
DP  - 2013
TI  - Sperm chromatin structure assay (SCSA(R)).
PG  - 147-64
AB  - The SCSA((R)) is the pioneering assay for the detection of damaged sperm DNA and 
      altered proteins in sperm nuclei via flow cytometry of acridine orange (AO) 
      stained sperm. The SCSA((R)) is considered to be the most precise and repeatable 
      test providing very unique, dual parameter data (red vs. green fluorescence) on a 
      1,024 x 1,024 channel scale, not only on DNA fragmentation but also on abnormal 
      sperm characterized by lack of normal exchange of histones to protamines. Raw 
      semen/sperm aliquots or purified sperm can be flash frozen, placed in a box with 
      dry ice and shipped by overnight courier to an experienced SCSA((R)) lab. The 
      samples are individually thawed, prepared, and analyzed in  approximately 10 min. Of 
      significance, data on 5,000 individual sperm are recorded on a 1,024 x 1,024 dot 
      plot of green (native DNA) and red (broken DNA) fluorescence. Repeat measurements 
      have virtually identical dot plot patterns demonstrating that the low pH 
      treatment that opens up the DNA strands at the sites of breaks and staining by 
      acridine orange (AO) are highly precise and repeatable (CVs of 1-3%) and the same 
      between fresh and frozen samples. SCSAsoft((R)) software transforms the X-Y data to 
      total DNA stainability versus red/red + green fluoresence (DFI) providing a more 
      accurate determination of % DFI as well as the more sensitive value of standard 
      deviation of DFI (SD DFI) as demonstrated by animal fertility and dose-response 
      toxicology studies. The current established clinical threshold is 25% DFI for 
      placing a man into a statistical probability of the following: (a) longer time to 
      natural pregnancy, (b) low odds of IUI pregnancy, (c) more miscarriages, or (d) 
      no pregnancy. Changes in lifestyle as well as medical intervention can lower the 
      %DFI to increase the probability of natural pregnancy. Couples of men with >25% 
      DFI are counseled to try ICSI and when in the >50% range may consider TESE/ICSI. 
      The SCSA((R)) simultaneously determines the % of sperm with high DNA stainability 
      (%HDS) related to retained nuclear histones consistent with immature sperm; high 
      HDS values are predictive of pregnancy failure.The SCSA((R)) is considered to be 
      the most technician friendly, time- and cost-efficient, precise and repeatable 
      DNA fragmentation assay, with the most data and the only fragmentation assay with 
      an accepted clinical threshold for placing a man at risk for infertility. SCSA((R)) 
      data are more predictive of male factor infertility than classical semen 
      analyses.
FAU - Evenson, Donald P
AU  - Evenson DP
AD  - SCSA Diagnostics, Volga, SD, USA. don@scsadiagnostics.com
LA  - eng
PT  - Journal Article
PL  - United States
TA  - Methods Mol Biol
JT  - Methods in molecular biology (Clifton, N.J.)
JID - 9214969
RN  - 0 (Chromatin)
RN  - 0 (Mutagens)
RN  - F30N4O6XVV (Acridine Orange)
SB  - IM
MH  - Acridine Orange
MH  - Animals
MH  - Chromatin/chemistry/drug effects/*metabolism
MH  - DNA Fragmentation/drug effects
MH  - Embryo Loss
MH  - Female
MH  - Flow Cytometry/*methods
MH  - Genes, Lethal
MH  - Humans
MH  - Infertility, Male/diagnosis/genetics/metabolism
MH  - Male
MH  - Mice
MH  - Mutagens
MH  - Mutation
MH  - Pregnancy
MH  - Reproducibility of Results
MH  - Semen Analysis/methods
MH  - Spermatozoa/drug effects/*metabolism
EDAT- 2012/09/21 06:00
MHDA- 2013/01/25 06:00
CRDT- 2012/09/21 06:00
PHST- 2012/09/21 06:00 [entrez]
PHST- 2012/09/21 06:00 [pubmed]
PHST- 2013/01/25 06:00 [medline]
AID - 10.1007/978-1-62703-038-0_14 [doi]
PST - ppublish
SO  - Methods Mol Biol. 2013;927:147-64. doi: 10.1007/978-1-62703-038-0_14.