{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,31]],"date-time":"2026-10-31T00:38:46Z","timestamp":1793407126205,"version":"4.5.0"},"reference-count":41,"publisher":"Frontiers Media SA","license":[{"start":{"date-parts":[[2024,7,19]],"date-time":"2024-07-19T00:00:00Z","timestamp":1721347200000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":["frontiersin.org"],"crossmark-restriction":true},"short-container-title":["Front. Cell. Infect. Microbiol."],"abstract":"<jats:sec><jats:title>Introduction<\/jats:title><jats:p>Human cytomegalovirus (HCMV) is the most common viral infection seen in newborns. The major route of transmission for acquired human cytomegalovirus infection is breast milk from mothers who are HCMV seropositive to the infants. Thus, a rapid, economical, and simple method to perform HCMV test in breast milk is crucial and necessary for preventing acquired HCMV infection, especially in underdeveloped regions with limited laboratory resources.<\/jats:p><\/jats:sec><jats:sec><jats:title>Methods<\/jats:title><jats:p>In this study, an effective technique for the detection of HCMV was constructed by combining multienzyme isothermal rapid amplification (MIRA) and lateral flow chromatography strip (LFD). Primers for the conserved HCMV sequence UL83 were utilized for MIRA-LFD testing.<\/jats:p><\/jats:sec><jats:sec><jats:title>Results<\/jats:title><jats:p>Our results showed that the entire MIRA reaction could be completed in 12 minutes at 37\u00b0C, and LFD outcomes could be observed visibly after 10 minutes. The detection sensitivity of this method reached 50 copy\/\u03bcl. Samples of breast milk were examined to compare MIRA-LFD and conventional qPCR. The accuracy of MIRA-LFD was 100%.<\/jats:p><\/jats:sec><jats:sec><jats:title>Discussion<\/jats:title><jats:p>The straightforward, rapid, economic features of the test can provide the significant advantages for the prevention of breast milk-acquired cytomegalovirus infection, particularly in resource-limited locations with high seroprevalence of cytomegalovirus.<\/jats:p><\/jats:sec>","DOI":"10.3389\/fcimb.2024.1430302","type":"journal-article","created":{"date-parts":[[2024,7,19]],"date-time":"2024-07-19T05:10:22Z","timestamp":1721365822000},"update-policy":"https:\/\/doi.org\/10.3389\/crossmark-policy","source":"Crossref","is-referenced-by-count":14,"title":["Rapid detection of human cytomegalovirus by multienzyme isothermal rapid amplification and lateral flow dipsticks"],"prefix":"10.3389","volume":"14","author":[{"given":"Ming-hui","family":"Liu","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Xiaochong","family":"Guo","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Mao-ling","family":"Sun","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jia-lun","family":"Li","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Shu-han","family":"Liu","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yun-zhou","family":"Chen","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Dong-yi","family":"Wang","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Lan","family":"Wang","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yu-zhang","family":"Li","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jun","family":"Yao","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yang","family":"Li","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yu-qing","family":"Pan","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1965","published-online":{"date-parts":[[2024,7,19]]},"reference":[{"key":"B1","doi-asserted-by":"publisher","first-page":"438","DOI":"10.1093\/cid\/ciy945","article-title":"Short-term pasteurization of breast milk to prevent postnatal cytomegalovirus transmission in very preterm infants","volume":"69","author":"Bapistella","year":"2019","journal-title":"Clin. Infect. diseases: an Off. Publ. Infect. Dis. Soc. America"},{"key":"B2","doi-asserted-by":"publisher","first-page":"679","DOI":"10.1007\/s00216-017-0620-3","article-title":"Helicase-dependent isothermal amplification: a novel tool in the development of molecular-based analytical systems for rapid pathogen detection","volume":"410","author":"Barreda-Garc\u00eda","year":"2018","journal-title":"Analytical bioanalytical Chem."},{"key":"B3","doi-asserted-by":"publisher","first-page":"1","DOI":"10.1146\/annurev-phyto-021621-115027","article-title":"Point-of-care DNA amplification for disease diagnosis and management","volume":"60","author":"Botella","year":"2022","journal-title":"Annu. Rev. Phytopathol."},{"key":"B4","doi-asserted-by":"publisher","DOI":"10.3389\/fcimb.2021.678703","article-title":"Rapid detection of SARS-CoV-2 using duplex reverse transcription-multienzyme isothermal rapid amplification in a point-of-care testing","volume":"11","author":"Chen","year":"2021","journal-title":"Front. Cell. infection Microbiol."},{"key":"B5","doi-asserted-by":"publisher","first-page":"30","DOI":"10.1016\/j.aca.2014.09.037","article-title":"Bioanalytical applications of isothermal nucleic acid amplification techniques","volume":"853","author":"Deng","year":"2015","journal-title":"Anal Chim Acta"},{"key":"B6","doi-asserted-by":"publisher","DOI":"10.1155\/2015\/218080","article-title":"Pregnancy outcomes of mothers with detectable CMV-specific IgM antibodies: a three-year review in a large irish tertiary referral maternity hospital","volume":"2015","author":"Drew","year":"2015","journal-title":"Infect. Dis. obstetrics gynecology"},{"key":"B7","doi-asserted-by":"publisher","DOI":"10.1128\/jcm.00283-23","article-title":"Multicenter evaluation of the diaSorin molecular simplexa congenital CMV direct PCR test on neonatal saliva and urine specimens","volume":"61","author":"Dunn","year":"2023","journal-title":"J. Clin. Microbiol."},{"key":"B8","doi-asserted-by":"publisher","first-page":"845","DOI":"10.1128\/JCM.01528-08","article-title":"Cross-priming amplification for rapid detection of Mycobacterium tuberculosis in sputum specimens","volume":"47","author":"Fang","year":"2009","journal-title":"J. Clin. Microbiol."},{"key":"B9","doi-asserted-by":"publisher","first-page":"660","DOI":"10.3390\/nu13020660","article-title":"Literature Review and an Italian Hospital Experience about Post-Natal CMV Infection Acquired by Breast-Feeding in Very Low and\/or Extremely Low Birth Weight Infants","volume":"13","author":"Garofoli","year":"2021","journal-title":"Nutrients"},{"key":"B10","doi-asserted-by":"publisher","first-page":"513","DOI":"10.1016\/S0140-6736(00)04043-5","article-title":"Epidemiology of transmission of cytomegalovirus from mother to preterm infant by breastfeeding","volume":"357","author":"Hamprecht","year":"2001","journal-title":"Lancet"},{"key":"B11","doi-asserted-by":"publisher","first-page":"757","DOI":"10.1016\/S0140-6736(67)91369-4","article-title":"Serum gamma-G-globulin levels and gestational age in premature babies","volume":"1","author":"Hobbs","year":"1967","journal-title":"Lancet"},{"key":"B12","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pbio.0040222","article-title":"DNA amplification and detection made simple (relatively)","volume":"4","author":"Hoff","year":"2006","journal-title":"PloS Biol."},{"key":"B13","doi-asserted-by":"publisher","first-page":"1475","DOI":"10.1586\/14737159.2015.1090877","article-title":"Recombinase polymerase amplification: Emergence as a critical molecular technology for rapid, low-resource diagnostics","volume":"15","author":"James","year":"2015","journal-title":"Expert Rev. Mol. diagnostics"},{"key":"B14","doi-asserted-by":"publisher","DOI":"10.3389\/fmolb.2022.853339","article-title":"Development of a polymerase spiral reaction-based isothermal assay for rapid identification of thrips palmi","volume":"9","author":"Jangra","year":"2022","journal-title":"Front. Mol. Biosci."},{"key":"B15","doi-asserted-by":"publisher","first-page":"660","DOI":"10.1016\/j.chom.2022.04.007","article-title":"Maternal immune protection against infectious diseases","volume":"30","author":"Langel","year":"2022","journal-title":"Cell Host Microbe"},{"key":"B16","doi-asserted-by":"publisher","first-page":"659","DOI":"10.1016\/j.foodchem.2018.07.010","article-title":"Inter-laboratory validation of visual loop-mediated isothermal amplification assays for GM contents screening","volume":"274","author":"Li","year":"2019","journal-title":"Food Chem."},{"key":"B17","doi-asserted-by":"publisher","first-page":"19","DOI":"10.1016\/j.trac.2017.10.015","article-title":"Recombinase polymerase amplification: Basics, applications and recent advances","volume":"98","author":"Lobato","year":"2018","journal-title":"Trends Analyt Chem."},{"key":"B18","doi-asserted-by":"publisher","first-page":"929","DOI":"10.1093\/cid\/ciw394","article-title":"Incidence, risk factors, and morbidity of acquired postnatal cytomegalovirus infection among preterm infants fed maternal milk in a highly seropositive population","volume":"63","author":"Martins-Celini","year":"2016","journal-title":"Clin. Infect. diseases: an Off. Publ. Infect. Dis. Soc. America"},{"key":"B19","doi-asserted-by":"publisher","first-page":"225","DOI":"10.1002\/jmv.10277","article-title":"Quantitation of human cytomegalovirus in recipients of solid organ transplants by real-time quantitative PCR and pp65 antigenemia","volume":"69","author":"Mengelle","year":"2003","journal-title":"J. Med. Virol."},{"key":"B20","doi-asserted-by":"publisher","first-page":"904","DOI":"10.1097\/INF.0000000000003671","article-title":"Breast milk and saliva for postnatal cyto\u2020megalovirus screening among very low birth weight infants","volume":"41","author":"Mukhopadhyay","year":"2022","journal-title":"Pediatr. Infect. Dis. J."},{"key":"B21","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pone.0227143","article-title":"Characterization of a universal screening approach for congenital CMV infection based on a highly-sensitive, quantitative, multiplex real-time PCR assay","volume":"15","author":"Nagel","year":"2020","journal-title":"PloS One"},{"key":"B22","doi-asserted-by":"publisher","DOI":"10.1542\/peds.2021-054272","article-title":"Promoting human milk and breastfeeding for the very low birth weight infant","volume":"148","author":"Parker","year":"2021","journal-title":"Pediatrics"},{"key":"B23","doi-asserted-by":"publisher","first-page":"64","DOI":"10.1002\/ana.410310112","article-title":"Lethal cytomegalovirus infection in preterm infants: clinical, radiological, and neuropathological findings","volume":"31","author":"Perlman","year":"1992","journal-title":"Ann. Neurol."},{"key":"B24","doi-asserted-by":"publisher","DOI":"10.1542\/peds.2021-055368","article-title":"An ethical analysis of newborn congenital cytomegalovirus screening","volume":"149","author":"Pesch","year":"2022","journal-title":"Pediatrics"},{"key":"B25","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pbio.0040204","article-title":"DNA detection using recombination proteins","volume":"4","author":"Piepenburg","year":"2006","journal-title":"PloS Biol."},{"key":"B26","doi-asserted-by":"publisher","first-page":"Cd002971","DOI":"10.1002\/14651858.CD002971.pub4","article-title":"Formula versus donor breast milk for feeding preterm or low birth weight infants","volume":"6","author":"Quigley","year":"2018","journal-title":"Cochrane Database systematic Rev."},{"key":"B27","doi-asserted-by":"publisher","first-page":"63","DOI":"10.1007\/s00430-015-0427-9","article-title":"Incidence of CMV co-infection in HIV-positive women and their neonates in a tertiary referral center: a cohort study","volume":"205","author":"Reitter","year":"2016","journal-title":"Med. Microbiol. Immunol."},{"key":"B28","doi-asserted-by":"publisher","first-page":"845","DOI":"10.1002\/jmv.v87.5","article-title":"Evaluation of cytomegalovirus infection in low-birth weight children by breast milk using a real-time polymerase chain reaction assay","volume":"87","author":"Romero-G\u00f3mez","year":"2015","journal-title":"J. Med. Virol."},{"key":"B29","doi-asserted-by":"publisher","DOI":"10.1002\/rmv.2399","article-title":"A systematic review and meta-analysis of the prevalence of human cytomegalovirus shedding in seropositive pregnant women","volume":"32","author":"Sapuan","year":"2022","journal-title":"Rev. Med. Virol."},{"key":"B30","doi-asserted-by":"publisher","first-page":"73","DOI":"10.1002\/rmv.484","article-title":"Acquisition of human cytomegalovirus infection in infants via breast milk: natural immunization or cause for concern","volume":"16","author":"Schleiss","year":"2006","journal-title":"Rev. Med. Virol."},{"key":"B31","doi-asserted-by":"publisher","first-page":"267","DOI":"10.1016\/j.jpeds.2016.10.024","article-title":"Congenital cytomegalovirus among children with cerebral palsy","volume":"181","author":"Smithers-Sheedy","year":"2017","journal-title":"J. Pediatr."},{"key":"B32","doi-asserted-by":"publisher","first-page":"1073","DOI":"10.1056\/NEJM198005083021908","article-title":"Breast milk and the risk of cytomegalovirus infection","volume":"302","author":"Stagno","year":"1980","journal-title":"New Engl. J. Med."},{"key":"B33","doi-asserted-by":"publisher","DOI":"10.3389\/fmolb.2021.763079","article-title":"Simple and feasible detection of hepatitis B virus via combination of multienzyme isothermal rapid amplification and lateral flow dipstick strip","volume":"8","author":"Sun","year":"2021","journal-title":"Front. Mol. Biosci."},{"key":"B34","doi-asserted-by":"publisher","DOI":"10.3389\/fcimb.2022.1004183","article-title":"Real-time multiple cross displacement amplification assay for rapid and sensitive detection of Hemophilus influenzae","volume":"12","author":"Sun","year":"2022","journal-title":"Front. Cell. infection Microbiol."},{"key":"B35","doi-asserted-by":"publisher","first-page":"233","DOI":"10.1080\/21691401.2023.2203198","article-title":"Simple and feasible detection of hepatitis a virus using reverse transcription multienzyme isothermal rapid amplification and lateral flow dipsticks without standard PCR laboratory","volume":"51","author":"Sun","year":"2023","journal-title":"Artif. cells nanomedicine Biotechnol."},{"key":"B36","doi-asserted-by":"publisher","first-page":"154","DOI":"10.24875\/rmimss.M20000013","article-title":"[Detection of cytomegalovirus in plasma of patient\u2019s HIV by duplex real time PCR]","volume":"58","author":"Valdez-Salazar","year":"2020","journal-title":"Rev. Med. del Instituto Mexicano del Seguro Soc."},{"key":"B37","doi-asserted-by":"publisher","first-page":"2378","DOI":"10.1128\/JCM.06800-11","article-title":"Clinical significance of low cytomegalovirus DNA levels in human plasma","volume":"50","author":"Waggoner","year":"2012","journal-title":"J. Clin. Microbiol."},{"key":"B38","doi-asserted-by":"publisher","DOI":"10.1016\/j.aca.2022.339479","article-title":"Nicking enzyme-free strand displacement amplification-assisted CRISPR-Cas-based colorimetric detection of prostate-specific antigen in serum samples","volume":"1195","author":"Wang","year":"2022","journal-title":"Analytica chimica Acta"},{"key":"B39","doi-asserted-by":"publisher","first-page":"970","DOI":"10.1039\/c3mb70304e","article-title":"Isothermal amplified detection of DNA and RNA","volume":"10","author":"Yan","year":"2014","journal-title":"Mol. Biosyst."},{"key":"B40","doi-asserted-by":"publisher","first-page":"1333","DOI":"10.1542\/peds.111.6.1333","article-title":"Evaluation of cytomegalovirus infections transmitted via breast milk in preterm infants with a real-time polymerase chain reaction assay","volume":"111","author":"Yasuda","year":"2003","journal-title":"Pediatrics"},{"key":"B41","doi-asserted-by":"publisher","DOI":"10.1002\/rmv.2034","article-title":"Estimation of the worldwide seroprevalence of cytomegalovirus: a systematic review and meta-analysis","volume":"29","author":"Zuhair","year":"2019","journal-title":"Rev. Med. Virol."}],"container-title":["Frontiers in Cellular and Infection Microbiology"],"original-title":[],"link":[{"URL":"https:\/\/www.frontiersin.org\/articles\/10.3389\/fcimb.2024.1430302\/full","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2024,7,19]],"date-time":"2024-07-19T05:10:29Z","timestamp":1721365829000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.frontiersin.org\/articles\/10.3389\/fcimb.2024.1430302\/full"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2024,7,19]]},"references-count":41,"alternative-id":["10.3389\/fcimb.2024.1430302"],"URL":"https:\/\/doi.org\/10.3389\/fcimb.2024.1430302","relation":{},"ISSN":["2235-2988"],"issn-type":[{"value":"2235-2988","type":"electronic"}],"subject":[],"published":{"date-parts":[[2024,7,19]]},"article-number":"1430302"}}