Terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) assay using bench top flow cytometer for evaluation of sperm DNA fragmentation in fertility laboratories: protocol, reference values, and quality control

Journal of Assisted Reproduction and Genetics
Rakesh SharmaAshok Agarwal

Abstract

The purpose of this study is to provide a detailed protocol and quality control steps for measuring sperm DNA fragmentation (SDF) by terminal deoxynucleotidyl transferase deoxyuridine triphosphate (dUTP) nick end labeling (TUNEL) assay using a new bench top flow cytometer, determine the reference value of SDF, and assess sensitivity, specificity, and distribution of SDF in infertile men and controls with proven and unproven fertility. Semen specimens from 95 controls and 261 infertile men referred to a male infertility testing laboratory were tested for SDF by TUNEL assay using Apo-Direct kit and a bench top flow cytometer. Percentage of cells positive for TUNEL was calculated. Inter- and intraobserver variability was examined. TUNEL cutoff value, sensitivity, specificity, and distribution of different cutoff values in controls and infertile patients were calculated. The reference value of SDF by TUNEL assay was 16.8 % with a specificity of 91.6 % and sensitivity of 32.6 %. The positive and negative predictive values were 91.4 and 33.1 %, respectively. The upper limit of DNA damage in infertile men was significantly higher (68.9 %) than that in the controls (19.6 %). TUNEL assay using flow cytometry is a reproducible and easy m...Continue Reading

References

Sep 26, 2001·Reproduction : the Official Journal of the Society for the Study of Fertility·R J Aitken, C Krausz
Jul 26, 2002·Fertility and Sterility·Ramadan A SalehRakesh K Sharma
May 2, 2003·Human Reproduction·Mehdi BenchaibJean François Guérin
Apr 7, 2004·Fertility and Sterility·Ralf HenkelThinus F Kruger
Aug 9, 2005·Human Reproduction·M SergerieG Bleau
Oct 11, 2005·Fertility and Sterility·José Luis FernándezChristopher De Jonge
Dec 9, 2008·Journal of Andrology·Armand Zini, Mark Sigman
Aug 1, 2009·Urology·Sergey I MoskovtsevJ Brendan M Mullen
Sep 19, 2009·Methods in Molecular Biology·Ashok AgarwalRakesh K Sharma
Dec 17, 2009·Journal of Assisted Reproduction and Genetics·Ryan T SchulteGary D Smith
Jun 25, 2010·Urology·Rakesh K SharmaAshok Agarwal
Aug 20, 2010·Systems Biology in Reproductive Medicine·Monica MuratoriElisabetta Baldi
Oct 5, 2010·International Journal of Urology : Official Journal of the Japanese Urological Association·Mary K SamplaskiEdmund Sabanegh
Jan 24, 2012·Fertility and Sterility·Gulfam AhmadLouis Bujan
Sep 21, 2012·Methods in Molecular Biology·Rakesh SharmaAshok Agarwal
Aug 13, 2013·The Journal of Urology·Ashok AgarwalMark Sigman
Aug 21, 2013·Reproductive Biomedicine Online·Sheena E M LewisParviz Gharagozloo
Apr 29, 2014·International Urology and Nephrology·Sandro C EstevesAshok Agarwal
Nov 21, 2014·Reproductive Biology and Endocrinology : RB&E·Saad AlshahraniEdmund Sabanegh
Dec 3, 2014·Fertility and Sterility·Gianpiero D PalermoZev Rosenwaks
Jun 23, 2015·Reproductive Biomedicine Online·Sheena E M Lewis

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Citations

Jul 18, 2016·Journal of Assisted Reproduction and Genetics·Sandro C Esteves
Jul 6, 2017·Cytometry. Part a : the Journal of the International Society for Analytical Cytology·E A ArifulinE V Sheval
Jul 24, 2018·Systems Biology in Reproductive Medicine·Xiao ShiTin-Chiu Li
Oct 14, 2017·Clinical and Experimental Reproductive Medicine·Hee-Jun ChiHum-Dai Park
Mar 12, 2019·Andrologia·Saleem Ali Banihani, Alayna'-Almarddyah Abdullah Al-Khawalde
Apr 17, 2019·Panminerva Medica·Ahmad MajzoubSandro C Esteves
May 2, 2018·Arab Journal of Urology·Manesh Kumar Panner Selvam, Ashok Agarwal
Nov 9, 2017·Current Protocols in Toxicology·Sajal GuptaAshok Agarwal
Oct 12, 2019·Zygote : the Biology of Gametes and Early Embryos·Thais Rose Dos Santos Hamilton, Mayra Elena Ortiz D'Ávila Assumpção
Jul 15, 2020·Andrologia·Marlon Martinez, Ahmad Majzoub
Aug 20, 2020·Andrologia·Kathy A RobertAshok Agarwal
May 28, 2020·Andrologia·Cătălina BarbăroșieRalf Henkel
Jul 25, 2020·Andrologia·Rakesh SharmaRalf Henkel
Mar 16, 2018·International Urology and Nephrology·Matheus Roque, Sandro C Esteves
Mar 4, 2020·Scientific Reports·Ming-Wen Li, K C Kent Lloyd
Sep 7, 2020·Food and Chemical Toxicology : an International Journal Published for the British Industrial Biological Research Association·Alexandra K TsarouchaConstantinos Simopoulos
Jun 5, 2020·ACS Synthetic Biology·Jasmine Puay Suan ChuaSaurabh Nirantar
May 4, 2021·Human Fertility : Journal of the British Fertility Society·Fatemeh AsgariZahra Zandieh

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