Clinical utility of sperm DNA fragmentation tests Aboubakr Elnashar Benha University Hospital, Egypt

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  • Clinical utility of sperm DNA

    fragmentation tests Aboubakr Elnashar

    Benha University Hospital, Egypt

    Aboubakr Elnashar

  • CONTENTS 1. Origin of SDF

    2. Etiology of SDF

    3. Assessment of SDF

    4. Correlation between semen parameters &

    SDF

    5. Correlation between clinical parameters &

    SDF

    6. Clinical utility of SDF tests

    2013

    2014

    2015

    Conclusion

    6 Aboubakr Elnashar

  • 1. ORIGIN OF SDF (Marin et al, 2012)

    First line:

    Spermiogenesis/or spermatogenesis

    A breakdown in keys of cell system:

    -Apoptosis

    -DNA repair

    -Chromatin remodelin

    Second line:

    Increase radical oxygen species

    Failure of antioxidant defense system during transport

    through male reproductive tract Aboubakr Elnashar

  • 2. ETIOLOGY OF SDF (Evgini et al, 2014)

    I. Primary Testicular Factors

    1. Abnormal germ cell apoptosis

    2. Advanced paternal age

    3. Exposure to gonadotoxins

    4. Sperm protamine deficiency

    5. High level of ROS

    II. Extratesticular Factors

    1. Smoking

    2. Radiation

    3. Chemotherapy

    4. Genital tract inflammation

    5. Testicular hyperthermia

    6. Varicocele Aboubakr Elnashar

  • 3. ASSESSMENT OF SDF

    Test Principle Method

    TUNEL

    ISNT

    Incorporation of probes

    at the site of damage

    Direct

    SCSA

    SCD

    Comet

    Susceptibility of DBs to

    denature in acid

    solution

    Indirect

    Aniline blue

    Toluidine blue

    Incorporation of probes

    to nuclear proteins

    Chromatin

    incorporation

    (Feijo and Esteves, 2014)

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  • Aboubakr Elnashar

  • Normal= 10

    Fragmented= 4

    DFI= 4X100/10+4

    =28.5%

    normal

    normal

    normal

    normal

    normal

    normal

    normal

    normal

    normal

    fragmented

    fragmented

    fragmented

    fragmented

    normal

    30: male infertility 15-30: RM.

    15: Excellent to Good fertility potential

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  • 4. CORRELATION BETWEEN SEMEN

    PARAMETERS & SDF

    Majority of the studies: an inverse correlation between SDF rate and sperm

    quality (sperm concentration, motility, vitality and

    morphology), irrespective of the age of the subjects (Lopez et al, 2013; Evgini et al, 2014)

    A significant negative correlation between % of morphologically normal spermatozoa and SDF

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  • 5. CORRELATION BETWEEN

    CLINICAL PARAMETERS & SDF

    SDF showing correlations with 1. Fertilization rates

    2. Embryonic development

    3. Implantation

    4. Pregnancy

    5. Abortion rates

    6. Congenital anomalies of the offspring (Evgini et al, 2014)

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  • 6. CLINICAL UTILITY OF SDF TESTS

    ASRM, 2013

    5 questions

    1. Does SDF test predict male fertility with natural

    conception?

    There is an association with increased SDF and reduced fertility in men

    (Simon, Lewis; 2011)

    Insufficient evidence (Level C) to use the test as a predictor of fertility {cut-points have not been clearly

    established and validated}

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  • 2. Does SDF test predict pregnancy with IUI?

    Conflicting A Level II-1study showed a positive predictive value of the SCSA test with DFI 30% associated with a lower PR and LBR. (Bungum et al, 2007)

    Other studies did not confirm the cutoff Another study found no association with DNA integrity and pregnancy with IUI.

    Insufficient evidence (Level C) to recommend the use of SDF tests to predict pregnancy with IUI.

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  • 3. Is SDF predictive of pregnancy with in IVF?

    Meta-analysis (Zini et al, 2011) SDF was associated with a modest but significant.

    reduction in IVF. PR

    Insufficient evidence (Level C) to recommend routine use of SDF testing for patients undergoing

    IVF.

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  • 4. Is SDF predictive of pregnancy with IVF and

    ICSI?

    A meta-analysis (Collins et al, 2008) SDF was significantly associated with PR in IVF/ICSI cycles.

    Association was mild Predictive ability was weak. Test cut-offs were not clearly established.

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  • Meta-analysis [Zini et al, 2011] No consistent relationship between SDF and embryo quality and/or development. The influence of SDF on embryo quality/development may be more significant in ICSI compared to IVF cycles.

    Insufficient evidence (Level C) to recommend routine DNA integrity testing for patients undergoing

    IVF/ICSI.

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  • 5. Is SDF predictive of pregnancy loss?

    A meta-analysis (Zini, 2008) significant association between SDF and pregnancy

    loss after IVF or ICSI

    Insufficient evidence (Level C) to recommend routine DNA integrity testing to predict

    pregnancy loss.

    For diagnostic test 1. Results must be reproducible

    2. Applicable to a given patient

    3. Change management of patient

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  • II. British Fertility Society, 2013

    Testing of SDF shows much promise both as diagnostic test for male infertility and

    prognostic test for ART outcomes.

    SDF closely associated with fertility outcomes including

    Negative relationships with: fertilization, embryo quality, implantation

    Positive relationships with: miscarriage and childhood diseases.

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  • III. Evgni et al, 2014: Clinical indications

    for SDF tests 1.Prolonged idiopathic infertility

    2.Low fertilization rate or bad quality embryos in

    IVF

    3. Implantation failure following IVF

    4.Repeated abortions

    5.Prolonged exposure to toxic environmental

    conditions affecting fertility

    6.Conventional seminal parameters found below

    the reference range

    7.Advanced male partner age

    8.Varicocele patients

    9.Cancer patients

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  • Semen analysis

    Abnormal

    SDF T

    DFI30

    ICSI

    DFI30

    Lifestyle, antioxidant

    SDF T after 3 months

    Normal+ Female factor

    ICSI Failed

    Due to its strong correlation with several aspects of ART

    procedures and further consequences for the offspring: SDF T

    should be integrated in routine clinical practice (Evgni et al, 2014)

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  • Aboubakr Elnashar

    Osman et al, 2015: SR and MA

    LBR increased significantly in couples with low SDF compared with those with high SDF

    After IVF and ICSI, men with low SDF had significantly higher LBR

    A sensitivity analysis: no statistically significant difference in LBR between low and high SDF when

    ICSI was used

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    Conclusion

    There is insufficient evidence to recommend the

    routine use of SDF testing in evaluation and

    treatment of infertile couple {level C}

    ?????????

    For diagnostic test

    1. Results must be reproducible

    2. Applicable to a given patient

    3. Change management of patient

  • Aboubakr Elnashar

    Aboubakr Elnashar Lectures

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