{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,7,7]],"date-time":"2026-07-07T15:09:02Z","timestamp":1783436942630,"version":"3.54.6"},"reference-count":10,"publisher":"Bentham Science Publishers Ltd.","issue":"1","license":[{"start":{"date-parts":[[2017,6,30]],"date-time":"2017-06-30T00:00:00Z","timestamp":1498780800000},"content-version":"unspecified","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/legalcode"}],"content-domain":{"domain":["openmedicinalchemistryjournal.com","benthamopen.com"],"crossmark-restriction":true},"short-container-title":["TOMCJ"],"published-print":{"date-parts":[[2017,6,30]]},"abstract":"\n Objectives:<\/jats:title>\n Normalize the quantification of residual DNA from Vero cells in the rabies vaccine for use in human VAHV I, by quantitative PCR in real time and the design of primers that amplified, highly repetitive sequences of Cercopithecus aethiops<\/jats:italic> and a constitutive gene according to sequences reported in the GenBank and quantifying the residual DNA in the vaccine VAHV I in three consecutive batches according to the standard set by the World Health Organization.<\/jats:p>\n <\/jats:sec>\n \n Methods:<\/jats:title>\n A real time quantitative method based on SYBR Green chemistry has been applied for the quantification of residual DNA (resDNA) using highly repetitive DNA (Alu) and a housekeeping gene (B-actin) as target sequences.<\/jats:p>\n <\/jats:sec>\n \n Results:<\/jats:title>\n The sensitivity achieved with this white sequence is within the reported limits and who are between 5 and 50 pg. For real time PCR optimization with Alu-p53, different concentrations of MgCl2 (0.5, 0.75, 1.0, 1.25 and 1.5 mm) in combination with three different concentrations of primers (75, 100 and 150nM) were used. pDNA in concentration of 1x107<\/jats:sup> copies \/ ul was used as template. Optimal concentrations were 1.25 mM MgCl2 and 100nM primers. To level of detection of 1.53 ng\/ul was found for p53-Alu and Alu-Glob and 0.39 ng\/ul for B-actin with gDNA curves.<\/jats:p>\n <\/jats:sec>\n \n Conclusion:<\/jats:title>\n Quantification of resDNA of vaccine VAHV I with close-ups of B-actin was normalized. Reached a sensitivity of 30 pg of resDNA\/dose VAHV I, with close-ups of B-actin. Found, in three consecutive batches, an amount less than 10 ng\/dose, these results suggest that the production process ensures vaccine resDNA removal, meeting international requirements for biological products for use in humans that use continuous cell lines for production.<\/jats:p>\n <\/jats:sec>","DOI":"10.2174\/1874104501711010066","type":"journal-article","created":{"date-parts":[[2017,7,12]],"date-time":"2017-07-12T02:21:12Z","timestamp":1499826072000},"page":"66-80","update-policy":"https:\/\/doi.org\/10.2174\/crossmark_policy","source":"Crossref","is-referenced-by-count":6,"title":["Standardization of DNA Residual Quantification Method of Vero Cell Rabies Vaccine for Human Use"],"prefix":"10.2174","volume":"11","author":[{"given":"Janeth del Carmen Arias","family":"Palacios","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Carlos Alberto Barrero","family":"Barreto","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jos\u00e9 Salvador Monta\u00f1a","family":"Lara","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"\u00c1ngela Mar\u00eda Londo\u00f1o","family":"Navas","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"965","reference":[{"key":"ref1","unstructured":"Belotto A, Schneider MC, Leanes LF, Correa E, Tamayo H, Fern\u00e1ndez E. \n Situaci\u00f3n actual de la rabia en Am\u00e9rica Latina. \n Organizaci\u00f3n Panamericana de la Salud 2003."},{"key":"ref2","unstructured":"Muniappan B, Thilly W. \n PCR assay to detect Vero cell DNA in vaccines. \n MIT. Cambridge, MA, USA : Center for Environmental Health Sciences and Division of Toxicology. 1996."},{"key":"ref3","first-page":"279","volume":"82","author":"Lovatt A.","year":"2002","unstructured":"Lovatt A. \n Applications of quantitative PCR in the biosafety and genetic stability assessment of biotechnology products. \n J Biotechnol \n 2002; \n 82\n (3)\n : 279-300.","journal-title":"J. Biotechnol."},{"key":"ref4","doi-asserted-by":"publisher","first-page":"123","DOI":"10.1006\/biol.2001.0283","volume":"29","author":"Lokteff M.","year":"2001","unstructured":"Lokteff M, Klinguer-Hamour C, Julien E, et al. \n Residual DNA quantification in clinical batches of BBG2Na, a recombinant subunit vaccine against human respiratory syncytial virus. \n Biologicals \n 2001; \n 29\n (2)\n : 123-32.","journal-title":"Biologicals"},{"key":"ref5","doi-asserted-by":"publisher","first-page":"511","DOI":"10.1016\/j.vaccine.2004.06.014","volume":"23","author":"Frazatti-Gallina N.M.","year":"2004","unstructured":"Frazatti-Gallina NM, Mour\u00e3o-Fuches RM, Paoli RL, et al. \n Vero-cell rabies vaccine produced using serum-free medium. \n Vaccine \n 2004; \n 23\n (4)\n : 511-7.","journal-title":"Vaccine"},{"key":"ref6","doi-asserted-by":"publisher","first-page":"53","DOI":"10.1016\/S0166-0934(03)00191-5","volume":"112","author":"Broberg E.K.","year":"2003","unstructured":"Broberg EK, Nyg\u00e5rdas M, Salmi AA, Hukkanen V. \n Low copy number detection of herpes simplex virus type 1 mRNA and mouse Th1 type cytokine mRNAs by Light Cycler quantitative real-time PCR. \n J Virol Methods \n 2003; \n 112\n (1-2)\n : 53-65.","journal-title":"J. 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Methods"},{"key":"ref7","doi-asserted-by":"publisher","unstructured":"Meuer S, Wittwer C, Nakagawara K, Wittwer K, Hahn M, Kaul K. \n Rapid Cycle Real-time PCR, Methods and Applications. \n Heidelberg: Springer Press 2001; pp. \n \n \n 21-34.","DOI":"10.1007\/978-3-642-59524-0"},{"key":"ref8","unstructured":"Green M, Sambrook J. \n Molecular Cloning: A Laboratory Manual. 4th ed. \n Cold Spring Harbor Laboratory 2014."},{"key":"ref9","first-page":"2947","volume":"21","author":"Thompson JD","year":"2007","unstructured":"Thompson JD, Higgins DC, Gibson TJ, Larkin MA, Blackshields G, Brown TJ. \n Clustal W and Clustal X version \n 20 Bioinformatics 23 \n 2007; \n 21\n : 2947-8.","journal-title":"2.0 Bioinformatics 23"},{"key":"ref10","doi-asserted-by":"publisher","first-page":"4673","DOI":"10.1093\/nar\/22.22.4673","volume":"22","author":"Thompson J.D.","year":"1994","unstructured":"Thompson JD, Higgins DG, Gibson TJ. \n CLUSTAL W: improving the sensitivity of progressive multiple sequence alignment through sequence weighting, position-specific gap penalties and weight matrix choice. \n Nucleic Acids Res \n 1994; \n 22\n (22)\n : 4673-80.","journal-title":"Nucleic Acids Res."}],"container-title":["The Open Medicinal Chemistry Journal"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/openmedicinalchemistryjournal.com\/contents\/volumes\/V11\/TOMCJ-11-66\/TOMCJ-11-66.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/openmedicinalchemistryjournal.com\/contents\/volumes\/V11\/TOMCJ-11-66\/TOMCJ-11-66.xml","content-type":"application\/xml","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/openmedicinalchemistryjournal.com\/contents\/volumes\/V11\/TOMCJ-11-66\/TOMCJ-11-66.pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2019,4,15]],"date-time":"2019-04-15T00:35:42Z","timestamp":1555288542000},"score":1,"resource":{"primary":{"URL":"https:\/\/openmedicinalchemistryjournal.com\/VOLUME\/11\/PAGE\/66\/"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2017,6,30]]},"references-count":10,"journal-issue":{"issue":"1","published-print":{"date-parts":[[2017,6,30]]}},"alternative-id":["TOMCJ-11-66"],"URL":"https:\/\/doi.org\/10.2174\/1874104501711010066","relation":{},"ISSN":["1874-1045"],"issn-type":[{"value":"1874-1045","type":"print"}],"subject":[],"published":{"date-parts":[[2017,6,30]]},"assertion":[{"value":"Peer Reviewed","order":0,"name":"review_status","label":"Review Status","group":{"name":"peer_review_details","label":"Peer Review Details"}},{"value":"Single blind","order":1,"name":"review_process","label":"Review Process","group":{"name":"peer_review_details","label":"Peer Review Details"}},{"value":"Checked with iThenticate","order":0,"name":"screening_status","label":"Screening Status","group":{"name":"plagiarism_screening","label":"Plagiarism Screening"}},{"value":"2017-01-14","order":0,"name":"received","label":"Received","group":{"name":"publication_history","label":"Publication History"}},{"value":"2017-03-30","order":1,"name":"revised","label":"Revised","group":{"name":"publication_history","label":"Publication History"}},{"value":"2017-04-11","order":2,"name":"accepted","label":"Accepted","group":{"name":"publication_history","label":"Publication History"}},{"value":"2017-06-30","order":3,"name":"published","label":"Published","group":{"name":"publication_history","label":"Publication History"}}]}}