{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,5]],"date-time":"2026-09-05T18:07:23Z","timestamp":1788631643441,"version":"build-2803163510"},"reference-count":47,"publisher":"Public Library of Science (PLoS)","issue":"11","license":[{"start":{"date-parts":[[2025,11,14]],"date-time":"2025-11-14T00:00:00Z","timestamp":1763078400000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"Laboratory of Medical Investigation (LIM) 52, Faculty of Medicine, University of S\u00c3\u00a3o Paulo, Brazil"}],"content-domain":{"domain":["www.plosone.org"],"crossmark-restriction":false},"short-container-title":["PLoS One"],"abstract":"<jats:sec id=\"sec001\">\n                    <jats:title>Background<\/jats:title>\n                    <jats:p>Data on the dynamics of Herpesviruses and Torque teno virus (TTV) in plasma from twin pregnancies are limited, despite their potential to provide insights into maternal immune status in this high-risk population.<\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec id=\"sec002\">\n                    <jats:title>Methods<\/jats:title>\n                    <jats:p>A convenience sample of plasma was obtained from 54 healthy women with twin pregnancies and no adverse outcomes. The plasma samples were collected at 8\u201315, 17\u201325, and 26\u201335 weeks of gestation at Careggi University Hospital, Florence, Italy. Real-time PCR detected Herpesviruses and TTV DNA, determined viral titers, and evaluated associations with cytokine levels and pregnancy-related parameters.<\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec id=\"sec003\">\n                    <jats:title>Results<\/jats:title>\n                    <jats:p>Herpesvirus were detected in 10 women (18.5%): three for herpes simplex virus (HSV)-1, three for human herpesvirus (HHV)-6, one for both HSV-1 and HHV-6, and one each for HSV-2, cytomegalovirus (CMV), or Epstein\u2013Barr virus (EBV). Only one woman was positive throughout all trimesters (HHV-6). TTV DNA was detected in 42 women (77.7%), persistently in 26 (48.1%) and transiently in 16 (29.6%). No associations were found between viral detection and plasma interleukin (IL)-1\u03b2, IL-6, tumor necrosis factor-\u03b1 (TNF-\u03b1), or pregnancy-related parameters, except for a higher prevalence of TTV in women with previous deliveries (p\u2009=\u20090.0402).<\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec id=\"sec004\">\n                    <jats:title>Conclusion<\/jats:title>\n                    <jats:p>The high frequency and variable viral titer of TTV with the transient circulation of some herpesviruses appear unaffected by the immunological adaptations inherent to this setting of twin pregnancies without adverse outcomes, as supported by the absence of correlation between inflammatory markers and viral dynamics. Nonetheless, ongoing monitoring of obstetric and postnatal outcomes remains warranted.<\/jats:p>\n                  <\/jats:sec>","DOI":"10.1371\/journal.pone.0335200","type":"journal-article","created":{"date-parts":[[2025,11,14]],"date-time":"2025-11-14T18:42:35Z","timestamp":1763145755000},"page":"e0335200","update-policy":"https:\/\/doi.org\/10.1371\/journal.pone.corrections_policy","source":"Crossref","is-referenced-by-count":2,"title":["Circulation of Herpesvirus and Alphatorquevirus DNA in each trimester in asymptomatic women pregnant with twins"],"prefix":"10.1371","volume":"20","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-5659-1052","authenticated-orcid":true,"given":"Tania Regina","family":"Tozetto-Mendoza","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Layla","family":"Honorato","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Mariane Pereira","family":"Brito","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Noely Evangelista","family":"Ferreira","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Iara M.","family":"Linhares","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Angela","family":"Silvano","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-5516-5177","authenticated-orcid":true,"given":"Antonio","family":"Charlys da Costa","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Silvia Helena","family":"Lima","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-8706-3925","authenticated-orcid":true,"given":"Viola","family":"Seravalli","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Mariarosaria","family":"Di Tommaso","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-5655-8108","authenticated-orcid":true,"given":"Maria Cassia","family":"Mendes-Correa","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Steven S.","family":"Witkin","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"340","published-online":{"date-parts":[[2025,11,14]]},"reference":[{"key":"pone.0335200.ref001","first-page":"21","article-title":"Classification of TTV and related viruses (anelloviruses)","volume":"331","author":"P Biagini","year":"2009","journal-title":"Curr Top Microbiol Immunol"},{"issue":"1","key":"pone.0335200.ref002","doi-asserted-by":"crossref","first-page":"121","DOI":"10.1006\/viro.2002.1545","article-title":"TT virus replicates in stimulated but not in nonstimulated peripheral blood mononuclear cells","volume":"301","author":"LF Mariscal","year":"2002","journal-title":"Virology"},{"key":"pone.0335200.ref003","doi-asserted-by":"crossref","first-page":"1283454","DOI":"10.3389\/fmed.2023.1283454","article-title":"Editorial: Torquetenovirus: predictive biomarker or innocent bystander in pathogenesis","volume":"10","author":"TR Tozetto-Mendoza","year":"2023","journal-title":"Front Med (Lausanne)"},{"key":"pone.0335200.ref004","doi-asserted-by":"crossref","first-page":"998","DOI":"10.3389\/fimmu.2020.00998","article-title":"Torque Teno Virus as a Potential Biomarker for Complications and Survival After Allogeneic Hematopoietic Stem Cell Transplantation","volume":"11","author":"A Pradier","year":"2020","journal-title":"Front Immunol"},{"issue":"4","key":"pone.0335200.ref005","doi-asserted-by":"crossref","first-page":"667","DOI":"10.1093\/infdis\/jiu209","article-title":"Torque Teno virus viremia correlates with intensity of maintenance immunosuppression in adult orthotopic liver transplant","volume":"210","author":"D Focosi","year":"2014","journal-title":"J Infect Dis"},{"issue":"11","key":"pone.0335200.ref006","doi-asserted-by":"crossref","first-page":"291","DOI":"10.5500\/wjt.v10.i11.291","article-title":"Torque teno virus in liver diseases and after liver transplantation","volume":"10","author":"A Mrzljak","year":"2020","journal-title":"World J Transplant"},{"issue":"5","key":"pone.0335200.ref007","doi-asserted-by":"crossref","first-page":"246","DOI":"10.1046\/j.1469-0691.2000.00069.x","article-title":"Transfusion-transmitted virus infection in HIV-1-seropositive patients","volume":"6","author":"M Debiaggi","year":"2000","journal-title":"Clin Microbiol Infect"},{"issue":"3","key":"pone.0335200.ref008","doi-asserted-by":"crossref","first-page":"334","DOI":"10.3390\/v17030334","article-title":"Torque Teno Virus: Lights and Shades","volume":"17","author":"P Brani","year":"2025","journal-title":"Viruses"},{"key":"pone.0335200.ref009","doi-asserted-by":"crossref","first-page":"1061230","DOI":"10.3389\/fcimb.2023.1061230","article-title":"The plasma virome in longitudinal samples from pregnant patients","volume":"13","author":"MJ Stout","year":"2023","journal-title":"Front Cell Infect Microbiol"},{"key":"pone.0335200.ref010","doi-asserted-by":"crossref","first-page":"1191938","DOI":"10.3389\/fmed.2023.1191938","article-title":"Maternal plasma and salivary anelloviruses in pregnancy and preterm birth","volume":"10","author":"C Kyathanahalli","year":"2023","journal-title":"Front Med (Lausanne)"},{"key":"pone.0335200.ref011","doi-asserted-by":"crossref","first-page":"998849","DOI":"10.3389\/fmicb.2022.998849","article-title":"Torquetenovirus in pregnancy: Correlation with vaginal microbiome, metabolome and pro-inflammatory cytokines","volume":"13","author":"S Morselli","year":"2022","journal-title":"Front Microbiol"},{"issue":"11","key":"pone.0335200.ref012","doi-asserted-by":"crossref","first-page":"2075","DOI":"10.1007\/s43032-020-00227-1","article-title":"Torquetenovirus Titer in Vaginal Secretions from Pregnant and Postpartum Women: Association with Absence of Lactobacillus crispatus and Levels of Lactic Acid and Matrix Metalloproteinase-8","volume":"27","author":"TR Tozetto-Mendoza","year":"2020","journal-title":"Reprod Sci"},{"key":"pone.0335200.ref013","doi-asserted-by":"crossref","first-page":"809312","DOI":"10.3389\/fmed.2021.809312","article-title":"The Torque Teno Virus Titer in Saliva Reflects the Level of Circulating CD4+ T Lymphocytes and HIV in Individuals Undergoing Antiretroviral Maintenance Therapy","volume":"8","author":"L Honorato","year":"2022","journal-title":"Front Med (Lausanne)"},{"issue":"1","key":"pone.0335200.ref014","doi-asserted-by":"crossref","first-page":"24691","DOI":"10.1038\/s41598-024-73870-2","article-title":"Torque teno virus: a potential marker of immune reconstitution in youths with vertically acquired HIV","volume":"14","author":"L Tarancon-Diez","year":"2024","journal-title":"Sci Rep"},{"issue":"8","key":"pone.0335200.ref015","doi-asserted-by":"crossref","first-page":"1470","DOI":"10.3390\/v13081470","article-title":"De Novo Polycomb Recruitment: Lessons from Latent Herpesviruses","volume":"13","author":"SA Dochnal","year":"2021","journal-title":"Viruses"},{"issue":"4","key":"pone.0335200.ref016","doi-asserted-by":"crossref","first-page":"590","DOI":"10.1016\/j.jcv.2014.10.002","article-title":"Cytomegalovirus DNAemia in pregnant women","volume":"61","author":"MG Revello","year":"2014","journal-title":"J Clin Virol"},{"issue":"5","key":"pone.0335200.ref017","doi-asserted-by":"crossref","first-page":"743","DOI":"10.1093\/cid\/ciy166","article-title":"Cytomegalovirus Shedding in Seropositive Pregnant Women From a High-Seroprevalence Population: The Brazilian Cytomegalovirus Hearing and Maternal Secondary Infection Study","volume":"67","author":"NG Barbosa","year":"2018","journal-title":"Clin Infect Dis"},{"key":"pone.0335200.ref018","doi-asserted-by":"crossref","first-page":"676988","DOI":"10.3389\/fcimb.2021.676988","article-title":"Vertical Transmission and Discordance of Cytomegalovirus in Twin Pregnancies","volume":"11","author":"J Hutton","year":"2021","journal-title":"Front Cell Infect Microbiol"},{"issue":"9","key":"pone.0335200.ref019","doi-asserted-by":"crossref","first-page":"1370","DOI":"10.3390\/v16091370","article-title":"Analysis of HSV1\/2 Infection Reveals an Association between HSV-2 Reactivation and Pregnancy","volume":"16","author":"S Dovrat","year":"2024","journal-title":"Viruses"},{"key":"pone.0335200.ref020","doi-asserted-by":"crossref","first-page":"48","DOI":"10.1016\/j.jcv.2018.04.016","article-title":"The risk of herpes simplex virus and human cytomegalovirus infection during pregnancy upon adverse pregnancy outcomes: A meta-analysis","volume":"104","author":"T-L Shi","year":"2018","journal-title":"J Clin Virol"},{"issue":"6","key":"pone.0335200.ref021","doi-asserted-by":"crossref","first-page":"3075","DOI":"10.3390\/ijerph18063075","article-title":"SARS-CoV-2 Infection and Inflammatory Response in a Twin Pregnancy","volume":"18","author":"A Trombetta","year":"2021","journal-title":"Int J Environ Res Public Health"},{"issue":"1","key":"pone.0335200.ref022","doi-asserted-by":"crossref","first-page":"10813","DOI":"10.1038\/s41598-024-61504-6","article-title":"Vaccination in twin pregnancies: comparison between immunization before conception and during pregnancy","volume":"14","author":"R Svirsky","year":"2024","journal-title":"Sci Rep"},{"key":"pone.0335200.ref023","doi-asserted-by":"crossref","first-page":"1634430","DOI":"10.3389\/fimmu.2025.1634430","article-title":"Hyporesponsive fetus-specific T cell responses in multiparous human pregnancy","volume":"16","author":"JC Morgan","year":"2025","journal-title":"Front Immunol"},{"key":"pone.0335200.ref024","doi-asserted-by":"crossref","first-page":"156668","DOI":"10.1016\/j.cyto.2024.156668","article-title":"Changes in cytokine and sequestosome-1 levels during twin pregnancy progression: Association with outcome","volume":"180","author":"A Silvano","year":"2024","journal-title":"Cytokine"},{"issue":"2","key":"pone.0335200.ref025","doi-asserted-by":"crossref","first-page":"311","DOI":"10.3201\/eid1002.030759","article-title":"Real-time reverse transcription-polymerase chain reaction assay for SARS-associated coronavirus","volume":"10","author":"SL Emery","year":"2004","journal-title":"Emerg Infect Dis"},{"issue":"24","key":"pone.0335200.ref026","doi-asserted-by":"crossref","first-page":"11999","DOI":"10.1128\/JVI.75.24.11999-12004.2001","article-title":"Dynamics of persistent TT virus infection, as determined in patients treated with alpha interferon for concomitant hepatitis C virus infection","volume":"75","author":"F Maggi","year":"2001","journal-title":"J Virol"},{"key":"pone.0335200.ref027","doi-asserted-by":"crossref","first-page":"1161091","DOI":"10.3389\/fmed.2023.1161091","article-title":"Characterization of Torquetenovirus in amniotic fluid at the time of in utero fetal surgery: correlation with early premature delivery and respiratory distress","volume":"10","author":"TR Tozetto-Mendoza","year":"2023","journal-title":"Front Med (Lausanne)"},{"issue":"3","key":"pone.0335200.ref028","doi-asserted-by":"crossref","first-page":"220","DOI":"10.1590\/0074-02760160354","article-title":"Diagnosis of human herpes virus 1 and 2 (HHV-1 and HHV-2): use of a synthetic standard curve for absolute quantification by real time polymerase chain reaction","volume":"112","author":"LRP Lima","year":"2017","journal-title":"Mem Inst Oswaldo Cruz"},{"issue":"4","key":"pone.0335200.ref029","doi-asserted-by":"crossref","first-page":"326","DOI":"10.1016\/j.jcv.2008.03.005","article-title":"Challenges in designing a Taqman-based multiplex assay for the simultaneous detection of Herpes simplex virus types 1 and 2 and Varicella-zoster virus","volume":"42","author":"M Weidmann","year":"2008","journal-title":"J Clin Virol"},{"issue":"1","key":"pone.0335200.ref030","doi-asserted-by":"crossref","first-page":"773","DOI":"10.1186\/s12879-019-4309-4","article-title":"Multiplex qPCR facilitates identification of betaherpesviruses in patients with acute liver failure of unknown etiology","volume":"19","author":"JV Raposo","year":"2019","journal-title":"BMC Infect Dis"},{"issue":"1","key":"pone.0335200.ref031","doi-asserted-by":"crossref","first-page":"1785801","DOI":"10.1080\/20002297.2020.1785801","article-title":"Longitudinal study on oral shedding of human betaherpesviruses 6 and 7 in renal transplant recipients reveals active replication","volume":"12","author":"JV Raposo","year":"2020","journal-title":"J Oral Microbiol"},{"issue":"3","key":"pone.0335200.ref032","doi-asserted-by":"crossref","first-page":"271","DOI":"10.1016\/j.bjid.2013.07.011","article-title":"Duplex realtime PCR method for Epstein-Barr virus and human DNA quantification: its application for post-transplant lymphoproliferative disorders detection","volume":"18","author":"MD Fellner","year":"2014","journal-title":"Braz J Infect Dis"},{"issue":"7","key":"pone.0335200.ref033","doi-asserted-by":"crossref","first-page":"3601","DOI":"10.1128\/JVI.78.7.3601-3620.2004","article-title":"Concurrent expression of latent and a limited number of lytic genes with immune modulation and antiapoptotic function by Kaposi\u2019s sarcoma-associated herpesvirus early during infection of primary endothelial and fibroblast cells and subsequent decline of lytic gene expression","volume":"78","author":"HH Krishnan","year":"2004","journal-title":"J Virol"},{"issue":"1","key":"pone.0335200.ref034","doi-asserted-by":"crossref","DOI":"10.23937\/2378-3648\/1410013","article-title":"Application of Synthetic Standard Curves for Absolute Quantification of Hepatitis A and E by Real-Time PCR","volume":"2","author":"RS Tourinho","year":"2015","journal-title":"J Genet Genome Res"},{"issue":"3","key":"pone.0335200.ref035","doi-asserted-by":"crossref","first-page":"144","DOI":"10.1002\/rmv.715","article-title":"Chromosomally integrated human herpesvirus 6: questions and answers","volume":"22","author":"PE Pellett","year":"2012","journal-title":"Rev Med Virol"},{"issue":"9","key":"pone.0335200.ref036","doi-asserted-by":"crossref","first-page":"1636","DOI":"10.1080\/14767058.2020.1763298","article-title":"Alphatorquevirus is the most prevalent virus identified in blood from a matched maternal-infant preterm cohort","volume":"35","author":"P Sloan","year":"2022","journal-title":"J Matern Fetal Neonatal Med"},{"issue":"2","key":"pone.0335200.ref037","doi-asserted-by":"crossref","first-page":"110556","DOI":"10.1016\/j.ygeno.2022.110556","article-title":"Comprehensive analysis of circulating viral DNA in maternal plasma at population-scale using low-pass whole-genome sequencing","volume":"115","author":"V Eldem","year":"2023","journal-title":"Genomics"},{"issue":"6","key":"pone.0335200.ref038","doi-asserted-by":"crossref","first-page":"2035","DOI":"10.1086\/315115","article-title":"Reactivation of human herpesvirus 6 during pregnancy","volume":"180","author":"H Dahl","year":"1999","journal-title":"J Infect Dis"},{"key":"pone.0335200.ref039","doi-asserted-by":"crossref","first-page":"942","DOI":"10.1016\/j.ajog.2005.05.081","article-title":"Differential expression of immune system-related components in midtrimester amniotic fluid from singleton and twin pregnancies","volume":"193","author":"SC Perni","year":"2005","journal-title":"Am J Obstet Gynecol"},{"issue":"9","key":"pone.0335200.ref040","doi-asserted-by":"crossref","DOI":"10.1097\/INF.0000000000004890","article-title":"Inherited Chromosomally Integrated HHV-6: Diagnosis and Clinical Features","volume":"44","author":"H Miura","year":"2025","journal-title":"Pediatr Infect Dis J"},{"issue":"7","key":"pone.0335200.ref041","doi-asserted-by":"crossref","first-page":"194","DOI":"10.3390\/v9070194","article-title":"Latency, Integration, and Reactivation of Human Herpesvirus-6","volume":"9","author":"SN Pantry","year":"2017","journal-title":"Viruses"},{"issue":"2","key":"pone.0335200.ref042","article-title":"Genomic Analyses from Non-invasive Prenatal Testing Reveal Genetic Associations, Patterns of Viral Infections, and Chinese Population History","volume":"175","author":"S Liu","year":"2018","journal-title":"Cell"},{"issue":"3","key":"pone.0335200.ref043","doi-asserted-by":"crossref","first-page":"199","DOI":"10.1111\/aji.12355","article-title":"Viral infections during pregnancy","volume":"73","author":"M Silasi","year":"2015","journal-title":"Am J Reprod Immunol"},{"issue":"2","key":"pone.0335200.ref044","first-page":"28","article-title":"Importance of chromosomally integrated HHV-6A and -6B in the diagnosis of active HHV-6 infection","volume":"15","author":"DA Clark","year":"2008","journal-title":"Herpes"},{"issue":"3","key":"pone.0335200.ref045","doi-asserted-by":"crossref","first-page":"513","DOI":"10.1542\/peds.2007-2838","article-title":"Chromosomal integration of human herpesvirus 6 is the major mode of congenital human herpesvirus 6 infection","volume":"122","author":"CB Hall","year":"2008","journal-title":"Pediatrics"},{"issue":"2","key":"pone.0335200.ref046","doi-asserted-by":"crossref","first-page":"104","DOI":"10.1016\/j.eimc.2020.04.003","article-title":"Absence of maternal HHV-6-specific T-cell response in the setting of human Herpesvirus-6 chromosomal integration: Potential impact on congenital infection","volume":"39","author":"I Machuca","year":"2021","journal-title":"Enferm Infecc Microbiol Clin (Engl Ed)"},{"issue":"1","key":"pone.0335200.ref047","doi-asserted-by":"crossref","first-page":"10201","DOI":"10.1038\/s41598-017-09857-z","article-title":"Influence of Pregnancy History on the Vaginal Microbiome of Pregnant Women in their First Trimester","volume":"7","author":"D Nasioudis","year":"2017","journal-title":"Sci Rep"}],"container-title":["PLOS One"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/dx.plos.org\/10.1371\/journal.pone.0335200","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,11,14]],"date-time":"2025-11-14T18:42:40Z","timestamp":1763145760000},"score":1,"resource":{"primary":{"URL":"https:\/\/dx.plos.org\/10.1371\/journal.pone.0335200"}},"subtitle":[],"editor":[{"given":"Simone","family":"Agostini","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"editor"}]}],"short-title":[],"issued":{"date-parts":[[2025,11,14]]},"references-count":47,"journal-issue":{"issue":"11","published-online":{"date-parts":[[2025,11,14]]}},"URL":"https:\/\/doi.org\/10.1371\/journal.pone.0335200","relation":{},"ISSN":["1932-6203"],"issn-type":[{"value":"1932-6203","type":"electronic"}],"subject":[],"published":{"date-parts":[[2025,11,14]]}}}