{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,5,5]],"date-time":"2026-05-05T00:47:33Z","timestamp":1777942053861,"version":"3.51.4"},"reference-count":41,"publisher":"MDPI AG","issue":"2","license":[{"start":{"date-parts":[[2023,1,5]],"date-time":"2023-01-05T00:00:00Z","timestamp":1672876800000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100001871","name":"Portuguese Foundation for Science and Technology\/MCTES","doi-asserted-by":"publisher","award":["UIDB\/00709\/2020"],"award-info":[{"award-number":["UIDB\/00709\/2020"]}],"id":[{"id":"10.13039\/501100001871","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100001871","name":"Portuguese Foundation for Science and Technology\/MCTES","doi-asserted-by":"publisher","award":["UIDP\/00709\/2020"],"award-info":[{"award-number":["UIDP\/00709\/2020"]}],"id":[{"id":"10.13039\/501100001871","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Diagnostics"],"abstract":"<jats:p>Cell-free DNA fragments detected in blood and in other biological fluids are released from apoptotic\/necrotic cells. In this study, we analyzed cfDNA levels in follicular fluid (FF) samples from patients with infertility. Samples were collected from 178 infertile women and cfDNA was extracted and quantified by qPCR, using ALU115 and ALU247 primers, and statistical correlations were performed. We found that cfDNA concentration was significantly higher in FF pools from women aged 35 and over than in women under 35 years of age (p = 0.017). We also found that q247 cfDNA levels were significantly higher in women with an associated female factor, such as endometriosis, PCOS and POF, compared with women with no specific cause of infertility (p = 0.033). The concentration of cfDNA did not vary significantly in each group of women with an associated female factor. The concentration of cfDNA was significantly higher in the FF of women that obtained embryos with a high fragmentation rate, compared to embryos with a low fragmentation rate (p = 0.007). Finally, we found that women who did not become pregnant during IVF treatments had higher q247 cfDNA levels (p = 0.043). The quantification of cfDNA could be an important biomarker of follicular micro-environment quality to predict embryo quality and the success of IVF, making them more specific and effective.<\/jats:p>","DOI":"10.3390\/diagnostics13020208","type":"journal-article","created":{"date-parts":[[2023,1,6]],"date-time":"2023-01-06T01:48:19Z","timestamp":1672969699000},"page":"208","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":9,"title":["Cell-Free DNA as a New Biomarker of IVF Success, Independent of Any Infertility Factor, Including Endometriosis"],"prefix":"10.3390","volume":"13","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-1823-2489","authenticated-orcid":false,"given":"Maria Manuel","family":"Casteleiro Alves","sequence":"first","affiliation":[{"name":"Health Sciences Research Centre (CICS), Faculty of Health Sciences, University of Beira Interior (UBI), 6201-506 Covilh\u00e3, Portugal"},{"name":"Assisted Reproduction Laboratory of Academic Hospital of Cova da Beira (CHUCB), 6200-251 Covilh\u00e3, Portugal"}]},{"given":"Lu\u00edsa","family":"Oliani","sequence":"additional","affiliation":[{"name":"School of Sciences, University of Minho, 4710-057 Braga, Portugal"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-4280-0765","authenticated-orcid":false,"given":"Micaela","family":"Almeida","sequence":"additional","affiliation":[{"name":"Health Sciences Research Centre (CICS), Faculty of Health Sciences, University of Beira Interior (UBI), 6201-506 Covilh\u00e3, Portugal"}]},{"given":"Henrique Jos\u00e9","family":"Cardoso","sequence":"additional","affiliation":[{"name":"Health Sciences Research Centre (CICS), Faculty of Health Sciences, University of Beira Interior (UBI), 6201-506 Covilh\u00e3, Portugal"}]},{"given":"Ant\u00f3nio H\u00e9lio","family":"Oliani","sequence":"additional","affiliation":[{"name":"Health Sciences Research Centre (CICS), Faculty of Health Sciences, University of Beira Interior (UBI), 6201-506 Covilh\u00e3, Portugal"},{"name":"Assisted Reproduction Laboratory of Academic Hospital of Cova da Beira (CHUCB), 6200-251 Covilh\u00e3, Portugal"},{"name":"Department of Obstetrics and Gynecology, S\u00e3o Jos\u00e9 do Rio Preto School of Medicine (FAMERP), S\u00e3o Jos\u00e9 do Rio Preto 15090-000, Brazil"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-8550-1645","authenticated-orcid":false,"given":"Luiza","family":"Breitenfeld","sequence":"additional","affiliation":[{"name":"Health Sciences Research Centre (CICS), Faculty of Health Sciences, University of Beira Interior (UBI), 6201-506 Covilh\u00e3, Portugal"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-4783-665X","authenticated-orcid":false,"given":"Ana Cristina","family":"Ramalhinho","sequence":"additional","affiliation":[{"name":"Health Sciences Research Centre (CICS), Faculty of Health Sciences, University of Beira Interior (UBI), 6201-506 Covilh\u00e3, Portugal"},{"name":"Assisted Reproduction Laboratory of Academic Hospital of Cova da Beira (CHUCB), 6200-251 Covilh\u00e3, Portugal"}]}],"member":"1968","published-online":{"date-parts":[[2023,1,5]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"1786","DOI":"10.1093\/humrep\/dex234","article-title":"The International Glossary on Infertility and Fertility Care","volume":"32","author":"Adamson","year":"2017","journal-title":"Hum. Reprod."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"481","DOI":"10.4103\/1008-682X.143737","article-title":"Association of polymorphisms in glutathione S-transferase genes (GSTM1, GSTT1, GSTP1) with idiopathic azoospermia or oligospermia in Sichuan, China","volume":"17","author":"Xiong","year":"2015","journal-title":"Asian J. Androl."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"148","DOI":"10.1016\/j.rbmo.2011.10.011","article-title":"Cryptic sperm defects may be the cause for total fertilization failure in oocyte donor cycles","volume":"24","author":"Goudakou","year":"2012","journal-title":"Reprod. Biomed. Online"},{"key":"ref_4","doi-asserted-by":"crossref","unstructured":"Gullo, G., Cucinella, G., Perino, A., Gullo, D., Segreto, D., Lagan\u00e0, A.S., Buzzaccarini, G., Donarelli, Z., Marino, A., and Allegra, A. (2021). The Gender Gap in the Diagnostic-Therapeutic Journey of the Infertile Couple. Int. J. Environ. Res. Public Health., 18.","DOI":"10.3390\/ijerph18126184"},{"key":"ref_5","doi-asserted-by":"crossref","unstructured":"Gardner, D., Sakkas, D., Seli, E., and Wells, D. (2013). Morphological assessment of blastocyst stage embryos: Types of grading systems and their reported outcomes. Human Gametes and Preimplantation Embryos: Assessment and Diagnosis, Springer Science and Media.","DOI":"10.1007\/978-1-4614-6651-2"},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"1497","DOI":"10.1007\/s10815-018-1223-4","article-title":"Relationship between blastocoel cell-free DNA and day-5 blastocyst morphology","volume":"35","author":"Rule","year":"2018","journal-title":"J. Assist. Reprod. Genet."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"279","DOI":"10.1016\/j.fertnstert.2006.06.025","article-title":"High concentrations of lactoferrin in the follicular fluid correlate with embryo quality during in vitro fertilization cycles","volume":"87","author":"Yanaihara","year":"2007","journal-title":"Fertil. Steril."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"1569","DOI":"10.1016\/j.fertnstert.2008.08.120","article-title":"A proteomic analysis of IVF follicular fluid in women \u226432 years old","volume":"92","author":"Estes","year":"2009","journal-title":"Fertil. Steril."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"40","DOI":"10.1186\/1477-7827-7-40","article-title":"Follicular fluid content and oocyte quality: From single biochemical markers to metabolomics","volume":"7","author":"Revelli","year":"2009","journal-title":"Reprod Biol. Endocrinol."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"500","DOI":"10.1016\/S1472-6483(10)62005-6","article-title":"Novel follicular fluid factors influencing oocyte developmental potential in IVF: A review","volume":"12","author":"Baka","year":"2006","journal-title":"Reprod. Biomed. Online."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"1078","DOI":"10.1016\/j.fertnstert.2012.01.122","article-title":"An investigation into the relationship between the metabolic profile of follicular fluid, oocyte developmental potential, and implantation outcome","volume":"97","author":"Wallace","year":"2012","journal-title":"Fertil. Steril."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"1775","DOI":"10.1016\/j.bbapap.2006.09.001","article-title":"Proteome analysis of human follicular fluid","volume":"1764","author":"Angelucci","year":"2006","journal-title":"Biochim. Et Biophys. Acta (BBA)-Proteins Proteom."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"707","DOI":"10.1016\/0277-5379(87)90266-5","article-title":"Isolation and characterization of DNA from the plasma of cancer patients","volume":"23","author":"Stroun","year":"1987","journal-title":"Eur. J. Cancer Clin. Oncol."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"898","DOI":"10.1158\/1541-7786.MCR-16-0044","article-title":"Cell-free DNA (cfDNA): Clinical Significance and Utility in Cancer Shaped by Emerging Technologies","volume":"14","author":"Volik","year":"2016","journal-title":"Mol. Cancer Res."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"2661","DOI":"10.1093\/humrep\/deu238","article-title":"Cell-free DNA in human follicular fluid as a biomarker of embryo quality","volume":"29","author":"Scalici","year":"2014","journal-title":"Hum Reprod."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"579","DOI":"10.1200\/JCO.2012.45.2011","article-title":"Liquid biopsies: Genotyping circulating tumor DNA","volume":"32","author":"Diaz","year":"2014","journal-title":"J. Clin. Oncol."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"426","DOI":"10.1038\/nrc3066","article-title":"Cell-free nucleic acids as biomarkers in cancer patients","volume":"11","author":"Schwarzenbach","year":"2011","journal-title":"Nat. Rev. Cancer."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"281","DOI":"10.1080\/08916930701358826","article-title":"The origin of extracellular DNA during the clearance of dead and dying cells","volume":"40","author":"Pisetsky","year":"2007","journal-title":"Autoimmunity."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"360","DOI":"10.2174\/187152612804142224","article-title":"Biology of circulating nucleic acids and possible roles in diagnosis and treatment in diabetes and cancer","volume":"12","author":"Gahan","year":"2012","journal-title":"Infect. Disord. Drug Targets."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"827","DOI":"10.1586\/14737159.2013.845088","article-title":"Diagnostic and prognostic value of circulating tumor-related DNA in cancer patients","volume":"13","author":"Marzese","year":"2013","journal-title":"Expert Rev. Mol. Diagn."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"485","DOI":"10.1016\/S0140-6736(97)02174-0","article-title":"Presence of fetal DNA in maternal plasma and serum","volume":"350","author":"Lo","year":"1997","journal-title":"Lancet."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"905","DOI":"10.1093\/humupd\/dmu031","article-title":"Cell-free nucleic acids as non-invasive biomarkers of gynecological cancers, ovarian, endometrial and obstetric disorders and fetal aneuploidy","volume":"20","author":"Traver","year":"2014","journal-title":"Hum. Reprod. Updat."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"593","DOI":"10.1016\/j.gyobfe.2015.07.016","article-title":"Les acides nucl\u00e9iques circulants et infertilit\u00e9 [Circulating nucleic acids and infertility]","volume":"43","author":"Scalici","year":"2015","journal-title":"Gyn\u00e9cologie Obs. Fertil."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"407","DOI":"10.1016\/S1472-6483(10)60100-9","article-title":"Circulating cell-free DNA as a potential biomarker for minimal and mild endometriosis","volume":"18","author":"Zachariah","year":"2009","journal-title":"Reprod. Biomed. Online."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"251","DOI":"10.1007\/s00737-013-0407-2","article-title":"Decrease in cell free DNA levels following participation in stress reduction techniques among women undergoing infertility treatment","volume":"17","author":"Sarid","year":"2014","journal-title":"Arch. Women's Ment. Health."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"e0136172","DOI":"10.1371\/journal.pone.0136172","article-title":"Cell-free DNA in Human Follicular Microenvironment: New Prognostic Biomarker to Predict in vitro Fertilization Outcomes","volume":"10","author":"Traver","year":"2015","journal-title":"PLoS ONE."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"169","DOI":"10.1016\/j.ejogrb.2012.03.003","article-title":"Injection of embryo culture supernatant to the endometrial cavity does not affect outcomes in IVF\/ICSI or oocyte donation cycles: A randomized clinical trial","volume":"162","author":"Prapas","year":"2012","journal-title":"Eur. J. Obstet. Gynecol. Reprod. Biol."},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"1062","DOI":"10.1373\/clinchem.2006.068577","article-title":"Increased integrity of free circulating DNA in sera of patients with colorectal or periampullary cancer: Direct quantitative PCR for ALU repeats","volume":"52","author":"Umetani","year":"2006","journal-title":"Clin. Chem."},{"key":"ref_29","first-page":"241","article-title":"Les acides nucl\u00e9iques du plasma sanguin chez l\u2019homme [Nuclear Acids in Human Blood Plasma]","volume":"142","author":"Mandel","year":"1948","journal-title":"CR Seances Soc. Biol. Fil."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"200","DOI":"10.3109\/10408363.2014.914888","article-title":"Nucleic acid-based biomarkers in body fluids of patients with urologic malignancies","volume":"51","author":"Ralla","year":"2014","journal-title":"Crit. Rev. Clin. Lab. Sci."},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"795","DOI":"10.1016\/j.febslet.2007.01.051","article-title":"Circulating (cell-free) nucleic acids--a promising, non-invasive tool for early detection of several human diseases","volume":"581","author":"Swarup","year":"2007","journal-title":"FEBS Lett."},{"key":"ref_32","first-page":"1659","article-title":"DNA fragments in the blood plasma of cancer patients: Quantitations and evidence for their origin from apoptotic and necrotic cells","volume":"61","author":"Jahr","year":"2001","journal-title":"Cancer Res."},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"47","DOI":"10.5114\/pm.2022.114404","article-title":"Psychological variables in medically assisted reproduction: A systematic review","volume":"21","author":"Burgio","year":"2022","journal-title":"Menopausal Rev."},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"124","DOI":"10.5114\/pm.2022.116437","article-title":"Impact of lifestyle and diet on endometriosis: A fresh look to a busy corner","volume":"21","author":"Habib","year":"2022","journal-title":"Menopausal Rev."},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"7265238","DOI":"10.1155\/2017\/7265238","article-title":"Oxidative Stress and Endometriosis: A Systematic Review of the Literature","volume":"2017","author":"Scutiero","year":"2017","journal-title":"Oxidative Med. Cell. Longev."},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"49","DOI":"10.1186\/1477-7827-10-49","article-title":"The effects of oxidative stress on female reproduction: A review","volume":"10","author":"Agarwal","year":"2012","journal-title":"Reprod. Biol. Endocrinol."},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"237","DOI":"10.1016\/j.exger.2006.01.004","article-title":"Mitochondrial respiration and reactive oxygen species in mitochondrial aging mutants","volume":"41","author":"Sedensky","year":"2006","journal-title":"Exp. Gerontol."},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"55","DOI":"10.1016\/j.ejmech.2015.04.040","article-title":"The role of antioxidants in the chemistry of oxidative stress: A review","volume":"97","author":"Pisoschi","year":"2015","journal-title":"Eur. J. Med. Chem."},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"367","DOI":"10.1093\/humupd\/dmn015","article-title":"Follicle dynamics and anovulation in polycystic ovary syndrome","volume":"14","author":"Franks","year":"2008","journal-title":"Hum. Reprod. Updat."},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"370","DOI":"10.1093\/humupd\/dmt062","article-title":"The physiology and clinical utility of anti-Mullerian hormone in women","volume":"20","author":"Dewailly","year":"2014","journal-title":"Hum. Reprod. Updat."},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"2652","DOI":"10.1093\/humrep\/det314","article-title":"Mitochondrial DNA content in embryo culture medium is significantly associated with human embryo fragmentation","volume":"28","author":"Stigliani","year":"2013","journal-title":"Hum. Reprod."}],"container-title":["Diagnostics"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2075-4418\/13\/2\/208\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,10]],"date-time":"2025-10-10T18:00:38Z","timestamp":1760119238000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2075-4418\/13\/2\/208"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2023,1,5]]},"references-count":41,"journal-issue":{"issue":"2","published-online":{"date-parts":[[2023,1]]}},"alternative-id":["diagnostics13020208"],"URL":"https:\/\/doi.org\/10.3390\/diagnostics13020208","relation":{},"ISSN":["2075-4418"],"issn-type":[{"value":"2075-4418","type":"electronic"}],"subject":[],"published":{"date-parts":[[2023,1,5]]}}}