PMID- 16242181
OWN - NLM
STAT- MEDLINE
DCOM- 20060509
LR  - 20220330
IS  - 0093-691X (Print)
IS  - 0093-691X (Linking)
VI  - 65
IP  - 5
DP  - 2006 Mar 15
TI  - Clinical aspects of sperm DNA fragmentation detection and male infertility.
PG  - 979-91
AB  - Over the past 25 years, various methods have been developed to measure sperm DNA 
      strand breaks in situ. Currently, there are four major tests of sperm DNA 
      fragmentation, including the Comet, Tunel, sperm chromatin structure assay (SCSA) 
      and the acridine orange test (AOT). The Comet assay is a light microscope 
      technique where the sperm cells are mixed with melted agarose and then placed on 
      a glass slide. The cells are lysed and then subjected to horizontal 
      electrophoresis. The Tunel assay, another light microscope technique, transfers 
      labeled nucleotide to the 3'OH group of a broken DNA strand with the use of 
      terminal deoxynucleotidyl transferase. The fluorescence intensity of each scored 
      sperm is determined as a "yes" or "no" for sperm on a light microscope slide or 
      by channels of fluorescent intensity in a flow cytometer. The light 
      microscope-based AOT, uses the metachromatic properties of acridine orange to 
      stain sperm cells. The SCSA treats sperm with low pH to denature DNA at the sites 
      of DNA strand breaks, followed by acridine orange (AO) staining of green for 
      native DNA and red for denatured DNA as measured by flow cytometry (FCM) as well 
      as % sperm with high DNA stainability (HDS: immature sperm with intact DNA 
      related to decreased fertilization rates). The SCSA method has defined a 27-30% 
      DNA fragmentation index (DFI) as the point in which a man is placed into a 
      statistical category of taking a longer time to in vivo pregnancy, intra uterine 
      insemination (IUI) and more routine in vitro fertilization (IVF) cycles or no 
      pregnancy. Current data suggest that intracytoplasmic sperm injection (ICSI) may 
      help overcome the diminished pregnancy prognosis with high DFI over the other ART 
      or natural methods.
FAU - Evenson, Donald P
AU  - Evenson DP
AD  - Department of Chemistry and Biochemistry, South Dakota State University, 
      Brookings, 57007, USA. scsa@brookings.net
FAU - Wixon, Regina
AU  - Wixon R
LA  - eng
PT  - Journal Article
PT  - Review
DEP - 20051019
PL  - United States
TA  - Theriogenology
JT  - Theriogenology
JID - 0421510
RN  - F30N4O6XVV (Acridine Orange)
SB  - IM
MH  - Acridine Orange
MH  - Animals
MH  - Comet Assay/veterinary
MH  - DNA Fragmentation/*physiology
MH  - Female
MH  - Flow Cytometry/veterinary
MH  - Humans
MH  - In Situ Nick-End Labeling/veterinary
MH  - Infertility, Male/diagnosis/*veterinary
MH  - Male
MH  - Pregnancy
MH  - Reproductive Techniques, Assisted/*veterinary
MH  - Sex Chromatin/ultrastructure
MH  - Spermatozoa/chemistry/*physiology/ultrastructure
RF  - 45
EDAT- 2005/10/26 09:00
MHDA- 2006/05/10 09:00
CRDT- 2005/10/26 09:00
PHST- 2005/10/26 09:00 [pubmed]
PHST- 2006/05/10 09:00 [medline]
PHST- 2005/10/26 09:00 [entrez]
AID - S0093-691X(05)00396-1 [pii]
AID - 10.1016/j.theriogenology.2005.09.011 [doi]
PST - ppublish
SO  - Theriogenology. 2006 Mar 15;65(5):979-91. doi: 
      10.1016/j.theriogenology.2005.09.011. Epub 2005 Oct 19.