Double-stranded sperm DNA fragmentation assessed using comet assay is associated with recurrent pregnancy loss

AJ Drakeley; RL Lunt; E Newton; RC Gregoire; L Watkins; K Bambang; AM Ellard; C Ledgerwood; L Rae; A Sloan; M Lawlor; L Poots; L Moore; Elizabeth Bailie; J Ribas-Maynou; Tara C. B. Moore

doi:10.1016/j.rbmo.2025.105184

Double-stranded sperm DNA fragmentation assessed using comet assay is associated with recurrent pregnancy loss

AJ Drakeley
, RL Lunt
, E Newton
, RC Gregoire
, L Watkins
, K Bambang
, AM Ellard
, C Ledgerwood
, L Rae
, A Sloan
, M Lawlor
, L Poots
, L Moore
, Elizabeth Bailie
, J Ribas-Maynou
, Tara C. B. Moore

Research output: Contribution to journal › Article › peer-review

2 Citations (Scopus)

23 Downloads (Pure)

Abstract

Research question
Do men attending a recurrent miscarriage clinic have high double-stranded sperm DNA damage compared with a sperm donor population?
Design
This prospective cohort study included 100 men attending a recurrent miscarriage clinic, and 81 sperm donors from a European sperm bank who had proven fertility. All semen samples were evaluated using the Examen Lab alkaline (Exact) comet assay, which identifies the global (single and double strand) DNA damage, and the neutral (Extend) comet assay, measuring only double-stranded sperm DNA fragmentation (dsSDF).
Results
Semen analysis showed that 66 male partners of women with a history of recurrent miscarriage were classified as being within normal parameters. Of these, however, 66 men (61%) had raised global SDF, and 52% had raised dsSDF. When evaluating each test separately, the Exact comet (global SDF) assay presented an area under the curve (AUC) of 0.690 (95% CI 0.623–0.756), with the neutral Extend comet (dsSDF only) assay having an AUC of 0.876 (95% CI 0.834–0.914), and the incidence of damage showed an improved AUC of 0.909 (95% CI 0.874–0.940).
Conclusions
This study of male factor SDF in a large cohort of men attending a recurrent miscarriage clinic, where they are rarely the focus of clinical investigation, shows a strong association with dsSDF and male factor-driven miscarriage contribution, highlighting the importance of male investigation in couples experiencing recurrent pregnancy loss.

Original language	English
Article number	105184
Pages (from-to)	1-33
Number of pages	33
Journal	Reproductive biomedicine online
Volume	52
Issue number	1
Early online date	4 Aug 2025
DOIs	https://doi.org/10.1016/j.rbmo.2025.105184
Publication status	Published (in print/issue) - 30 Jan 2026

Bibliographical note

Publisher Copyright:
© 2025 The Author(s)

Funding

The study was part-funded using an unrestricted medical educational grant provided by Merck Serono Limited (0111897641) to the Liverpool Women’s Hospital.

Funders	Funder number
Liverpool Women's Hospital
Merck	0111897641

UN SDGs

This output contributes to the following UN Sustainable Development Goals (SDGs)

SDG 3 Good Health and Well-being

Keywords

Recurrent miscarriage
comet assay
DNA fragmentation
Double-stranded DNA break
Comet assay
Prospective Studies
Humans
Semen Analysis
Male
Comet Assay
Pregnancy
Spermatozoa/metabolism
DNA Fragmentation
Female
Adult
Abortion, Habitual/genetics

Access to Document

10.1016/j.rbmo.2025.105184Licence: CC BY

PIIS1472648325003918Final published version, 1.88 MBLicence: CC BY

Cite this

Drakeley, AJ., Lunt, RL., Newton, E., Gregoire, RC., Watkins, L., Bambang, K., Ellard, AM., Ledgerwood, C., Rae, L., Sloan, A., Lawlor, M., Poots, L., Moore, L., Bailie, E., Ribas-Maynou, J., & Moore, T. C. B. (2026). Double-stranded sperm DNA fragmentation assessed using comet assay is associated with recurrent pregnancy loss. Reproductive biomedicine online, 52(1), 1-33. Article 105184. https://doi.org/10.1016/j.rbmo.2025.105184

@article{168bb43ec3454db69b57d4f2ab591382,

title = "Double-stranded sperm DNA fragmentation assessed using comet assay is associated with recurrent pregnancy loss",

abstract = "Research question Do men attending a recurrent miscarriage clinic have high double-stranded sperm DNA damage compared with a sperm donor population? Design This prospective cohort study included 100 men attending a recurrent miscarriage clinic, and 81 sperm donors from a European sperm bank who had proven fertility. All semen samples were evaluated using the Examen Lab alkaline (Exact) comet assay, which identifies the global (single and double strand) DNA damage, and the neutral (Extend) comet assay, measuring only double-stranded sperm DNA fragmentation (dsSDF). Results Semen analysis showed that 66 male partners of women with a history of recurrent miscarriage were classified as being within normal parameters. Of these, however, 66 men (61\%) had raised global SDF, and 52\% had raised dsSDF. When evaluating each test separately, the Exact comet (global SDF) assay presented an area under the curve (AUC) of 0.690 (95\% CI 0.623–0.756), with the neutral Extend comet (dsSDF only) assay having an AUC of 0.876 (95\% CI 0.834–0.914), and the incidence of damage showed an improved AUC of 0.909 (95\% CI 0.874–0.940). Conclusions This study of male factor SDF in a large cohort of men attending a recurrent miscarriage clinic, where they are rarely the focus of clinical investigation, shows a strong association with dsSDF and male factor-driven miscarriage contribution, highlighting the importance of male investigation in couples experiencing recurrent pregnancy loss.",

keywords = "Recurrent miscarriage, comet assay, DNA fragmentation, Double-stranded DNA break, Comet assay, Prospective Studies, Humans, Semen Analysis, Male, Comet Assay, Pregnancy, Spermatozoa/metabolism, DNA Fragmentation, Female, Adult, Abortion, Habitual/genetics",

author = "AJ Drakeley and RL Lunt and E Newton and RC Gregoire and L Watkins and K Bambang and AM Ellard and C Ledgerwood and L Rae and A Sloan and M Lawlor and L Poots and L Moore and Elizabeth Bailie and J Ribas-Maynou and Moore, \{Tara C. B.\}",

note = "Publisher Copyright: {\textcopyright} 2025 The Author(s)",

year = "2026",

month = jan,

day = "30",

doi = "10.1016/j.rbmo.2025.105184",

language = "English",

volume = "52",

pages = "1--33",

journal = "Reproductive biomedicine online",

issn = "1472-6491",

publisher = "Elsevier Ltd",

number = "1",

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Drakeley, AJ, Lunt, RL, Newton, E, Gregoire, RC, Watkins, L, Bambang, K, Ellard, AM, Ledgerwood, C, Rae, L, Sloan, A, Lawlor, M, Poots, L, Moore, L, Bailie, E, Ribas-Maynou, J & Moore, TCB 2026, 'Double-stranded sperm DNA fragmentation assessed using comet assay is associated with recurrent pregnancy loss', Reproductive biomedicine online, vol. 52, no. 1, 105184, pp. 1-33. https://doi.org/10.1016/j.rbmo.2025.105184

TY - JOUR

T1 - Double-stranded sperm DNA fragmentation assessed using comet assay is associated with recurrent pregnancy loss

AU - Drakeley, AJ

AU - Lunt, RL

AU - Newton, E

AU - Gregoire, RC

AU - Watkins, L

AU - Bambang, K

AU - Ellard, AM

AU - Ledgerwood, C

AU - Rae, L

AU - Sloan, A

AU - Lawlor, M

AU - Poots, L

AU - Moore, L

AU - Bailie, Elizabeth

AU - Ribas-Maynou, J

AU - Moore, Tara C. B.

PY - 2026/1/30

Y1 - 2026/1/30

N2 - Research question Do men attending a recurrent miscarriage clinic have high double-stranded sperm DNA damage compared with a sperm donor population? Design This prospective cohort study included 100 men attending a recurrent miscarriage clinic, and 81 sperm donors from a European sperm bank who had proven fertility. All semen samples were evaluated using the Examen Lab alkaline (Exact) comet assay, which identifies the global (single and double strand) DNA damage, and the neutral (Extend) comet assay, measuring only double-stranded sperm DNA fragmentation (dsSDF). Results Semen analysis showed that 66 male partners of women with a history of recurrent miscarriage were classified as being within normal parameters. Of these, however, 66 men (61%) had raised global SDF, and 52% had raised dsSDF. When evaluating each test separately, the Exact comet (global SDF) assay presented an area under the curve (AUC) of 0.690 (95% CI 0.623–0.756), with the neutral Extend comet (dsSDF only) assay having an AUC of 0.876 (95% CI 0.834–0.914), and the incidence of damage showed an improved AUC of 0.909 (95% CI 0.874–0.940). Conclusions This study of male factor SDF in a large cohort of men attending a recurrent miscarriage clinic, where they are rarely the focus of clinical investigation, shows a strong association with dsSDF and male factor-driven miscarriage contribution, highlighting the importance of male investigation in couples experiencing recurrent pregnancy loss.

AB - Research question Do men attending a recurrent miscarriage clinic have high double-stranded sperm DNA damage compared with a sperm donor population? Design This prospective cohort study included 100 men attending a recurrent miscarriage clinic, and 81 sperm donors from a European sperm bank who had proven fertility. All semen samples were evaluated using the Examen Lab alkaline (Exact) comet assay, which identifies the global (single and double strand) DNA damage, and the neutral (Extend) comet assay, measuring only double-stranded sperm DNA fragmentation (dsSDF). Results Semen analysis showed that 66 male partners of women with a history of recurrent miscarriage were classified as being within normal parameters. Of these, however, 66 men (61%) had raised global SDF, and 52% had raised dsSDF. When evaluating each test separately, the Exact comet (global SDF) assay presented an area under the curve (AUC) of 0.690 (95% CI 0.623–0.756), with the neutral Extend comet (dsSDF only) assay having an AUC of 0.876 (95% CI 0.834–0.914), and the incidence of damage showed an improved AUC of 0.909 (95% CI 0.874–0.940). Conclusions This study of male factor SDF in a large cohort of men attending a recurrent miscarriage clinic, where they are rarely the focus of clinical investigation, shows a strong association with dsSDF and male factor-driven miscarriage contribution, highlighting the importance of male investigation in couples experiencing recurrent pregnancy loss.

KW - Recurrent miscarriage

KW - comet assay

KW - DNA fragmentation

KW - Double-stranded DNA break

KW - Comet assay

KW - Prospective Studies

KW - Humans

KW - Semen Analysis

KW - Male

KW - Comet Assay

KW - Pregnancy

KW - Spermatozoa/metabolism

KW - DNA Fragmentation

KW - Female

KW - Adult

KW - Abortion, Habitual/genetics

UR - https://www.scopus.com/pages/publications/105024227301

U2 - 10.1016/j.rbmo.2025.105184

DO - 10.1016/j.rbmo.2025.105184

M3 - Article

C2 - 41371000

SN - 1472-6491

VL - 52

SP - 1

EP - 33

JO - Reproductive biomedicine online

JF - Reproductive biomedicine online

IS - 1

M1 - 105184

ER -

Double-stranded sperm DNA fragmentation assessed using comet assay is associated with recurrent pregnancy loss

Abstract

Bibliographical note

Funding

UN SDGs

Keywords

Access to Document

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