{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,11,2]],"date-time":"2026-11-02T19:31:08Z","timestamp":1793647868594,"version":"4.5.1"},"reference-count":60,"publisher":"Springer Science and Business Media LLC","issue":"1","license":[{"start":{"date-parts":[[2024,2,19]],"date-time":"2024-02-19T00:00:00Z","timestamp":1708300800000},"content-version":"tdm","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0"},{"start":{"date-parts":[[2024,2,19]],"date-time":"2024-02-19T00:00:00Z","timestamp":1708300800000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0"}],"funder":[{"DOI":"10.13039\/501100002347","name":"Bundesministerium f\u00fcr Bildung und Forschung","doi-asserted-by":"publisher","award":["13N15829"],"award-info":[{"award-number":["13N15829"]}],"id":[{"id":"10.13039\/501100002347","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100005721","name":"Universit\u00e4t Bielefeld","doi-asserted-by":"crossref","id":[{"id":"10.13039\/501100005721","id-type":"DOI","asserted-by":"crossref"}]}],"content-domain":{"domain":["link.springer.com"],"crossmark-restriction":false},"short-container-title":["Sci Rep"],"abstract":"<jats:title>Abstract<\/jats:title><jats:p>Molecular diagnostics is an increasingly important clinical tool, especially in routine sampling. We evaluated two non-invasive methods (oral swabs and mouthwashes) for sampling nucleic acids from the oral\/pharyngeal area. We created a workflow from sample collection (n\u2009=\u200959) to RT-qPCR based analysis. The samples were further characterized in terms of their cellular composition as well as the purity, degradation and microbial content of the derived DNA\/RNA. We determined the optimal housekeeping genes applicable for these types of samples. The cellular composition indicated that mouthwashes contained more immune cells and bacteria. Even though the protocol was not specifically optimized to extract bacterial RNA it was possible to derive microbial RNA, from both sampling methods. Optimizing the protocol allowed us to generate stable quantities of DNA\/RNA. DNA\/RNA purity parameters were not significantly different between the two sampling methods. Even though integrity analysis demonstrated a high level of degradation of RNA, corresponding parameters confirmed their sequencing potential. RT-qPCR analysis determined <jats:italic>TATA-Box Binding Protein<\/jats:italic> as the most favorable housekeeping gene. In summary, we have developed a robust method suitable for multiple downstream diagnostic techniques. This protocol can be used as a foundation for further research endeavors focusing on developing molecular diagnostics for the oropharyngeal cavity. <\/jats:p>","DOI":"10.1038\/s41598-024-54179-6","type":"journal-article","created":{"date-parts":[[2024,2,19]],"date-time":"2024-02-19T09:03:19Z","timestamp":1708333399000},"update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":9,"title":["Characterization of non-invasive oropharyngeal samples and nucleic acid isolation for molecular diagnostics"],"prefix":"10.1038","volume":"14","author":[{"given":"Leonie","family":"Hose","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Matthias","family":"Sch\u00fcrmann","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Inga","family":"Mennebr\u00f6cker","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Rayoung","family":"Kim","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Tobias","family":"Busche","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Peter","family":"Goon","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Holger","family":"Sudhoff","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"297","published-online":{"date-parts":[[2024,2,19]]},"reference":[{"key":"54179_CR1","doi-asserted-by":"publisher","first-page":"8761860","DOI":"10.1155\/2019\/8761860","volume":"2019","author":"A Roi","year":"2019","unstructured":"Roi, A. et al. A new approach for the diagnosis of systemic and oral diseases based on salivary biomolecules. Dis. Mark. 2019, 8761860. https:\/\/doi.org\/10.1155\/2019\/8761860 (2019).","journal-title":"Dis. Mark."},{"key":"54179_CR2","doi-asserted-by":"publisher","first-page":"6406","DOI":"10.1038\/s41598-021-85120-w","volume":"11","author":"N Bostanci","year":"2021","unstructured":"Bostanci, N. et al. Validation and verification of predictive salivary biomarkers for oral health. Sci. Rep. 11, 6406. https:\/\/doi.org\/10.1038\/s41598-021-85120-w (2021).","journal-title":"Sci. Rep."},{"key":"54179_CR3","doi-asserted-by":"publisher","first-page":"85","DOI":"10.1080\/14737159.2017.1404906","volume":"18","author":"LMRB Arantes","year":"2018","unstructured":"Arantes, L. M. R. B., de Carvalho, A. C., Melendez, M. E. & LopesCarvalho, A. Serum, plasma and saliva biomarkers for head and neck cancer. Expert Rev. Mol. Diagn. 18, 85\u2013112. https:\/\/doi.org\/10.1080\/14737159.2017.1404906 (2018).","journal-title":"Expert Rev. Mol. Diagn."},{"key":"54179_CR4","doi-asserted-by":"publisher","first-page":"949-57.e1-7","DOI":"10.1053\/j.gastro.2009.11.010","volume":"138","author":"L Zhang","year":"2010","unstructured":"Zhang, L. et al. Salivary transcriptomic biomarkers for detection of resectable pancreatic cancer. Gastroenterology 138, 949-57.e1\u20137. https:\/\/doi.org\/10.1053\/j.gastro.2009.11.010 (2010).","journal-title":"Gastroenterology"},{"key":"54179_CR5","first-page":"232","volume":"20","author":"GL McMichael","year":"2009","unstructured":"McMichael, G. L. et al. DNA from buccal swabs suitable for high-throughput SNP multiplex analysis. J. Biomol. Tech. JBT 20, 232\u2013235 (2009).","journal-title":"J. Biomol. Tech. JBT"},{"key":"54179_CR6","doi-asserted-by":"publisher","first-page":"148","DOI":"10.1590\/s0103-64402007000200012","volume":"18","author":"M Aidar","year":"2007","unstructured":"Aidar, M. & Line, S. R. P. A simple and cost-effective protocol for DNA isolation from buccal epithelial cells. Braz. Dent. J. 18, 148\u2013152. https:\/\/doi.org\/10.1590\/s0103-64402007000200012 (2007).","journal-title":"Braz. Dent. J."},{"issue":"262\u20134","key":"54179_CR7","doi-asserted-by":"publisher","first-page":"266","DOI":"10.2144\/04372RR03","volume":"37","author":"A Michalczyk","year":"2004","unstructured":"Michalczyk, A., Varigos, G., Smith, L. & Ackland, M. L. Fresh and cultured buccal cells as a source of mRNA and protein for molecular analysis. Biotechniques 37(262\u20134), 266\u2013269. https:\/\/doi.org\/10.2144\/04372RR03 (2004).","journal-title":"Biotechniques"},{"key":"54179_CR8","doi-asserted-by":"publisher","first-page":"127","DOI":"10.5858\/2001-125-0127-UOBCCI","volume":"125","author":"EM Heath","year":"2001","unstructured":"Heath, E. M. et al. Use of buccal cells collected in mouthwash as a source of DNA for clinical testing. Arch. Pathol. Lab. Med. 125, 127\u2013133. https:\/\/doi.org\/10.5858\/2001-125-0127-UOBCCI (2001).","journal-title":"Arch. Pathol. Lab. Med."},{"key":"54179_CR9","doi-asserted-by":"publisher","DOI":"10.1007\/978-90-481-3261-4","volume-title":"Molecular Diagnostics: Promises and Possibilities","author":"M Debnath","year":"2010","unstructured":"Debnath, M. & Bisen, P. S. Molecular Diagnostics: Promises and Possibilities 1st edn. (Springer, 2010).","edition":"1"},{"key":"54179_CR10","doi-asserted-by":"publisher","first-page":"745","DOI":"10.1111\/j.1601-0825.2011.01808.x","volume":"17","author":"RW Hart","year":"2011","unstructured":"Hart, R. W. et al. Point-of-care oral-based diagnostics. Oral Dis. 17, 745\u2013752. https:\/\/doi.org\/10.1111\/j.1601-0825.2011.01808.x (2011).","journal-title":"Oral Dis."},{"key":"54179_CR11","first-page":"152","volume":"8","author":"HY Liu","year":"2019","unstructured":"Liu, H. Y. et al. Polymerase chain reaction and its application in the diagnosis of infectious keratitis. Med. Hypothesis Discov. Innov. Ophthalmol. J. 8, 152\u2013155 (2019).","journal-title":"Med. Hypothesis Discov. Innov. Ophthalmol. J."},{"key":"54179_CR12","doi-asserted-by":"publisher","first-page":"112","DOI":"10.2144\/04371RR03","volume":"37","author":"K Dheda","year":"2004","unstructured":"Dheda, K. et al. Validation of housekeeping genes for normalizing RNA expression in real-time PCR. BioTechniques 37, 112\u2013114. https:\/\/doi.org\/10.2144\/04371RR03 (2004) (116, 118\u2013119).","journal-title":"BioTechniques"},{"key":"54179_CR13","doi-asserted-by":"publisher","first-page":"2312","DOI":"10.1038\/s41598-022-05670-5","volume":"12","author":"P Ostheim","year":"2022","unstructured":"Ostheim, P. et al. Examining potential confounding factors in gene expression analysis of human saliva and identifying potential housekeeping genes. Sci. Rep. 12, 2312. https:\/\/doi.org\/10.1038\/s41598-022-05670-5 (2022).","journal-title":"Sci. Rep."},{"key":"54179_CR14","doi-asserted-by":"publisher","first-page":"5245","DOI":"10.1158\/0008-5472.CAN-04-0496","volume":"64","author":"CL Andersen","year":"2004","unstructured":"Andersen, C. L., Jensen, J. L. & \u00d8rntoft, T. F. Normalization of real-time quantitative reverse transcription-PCR data: A model-based variance estimation approach to identify genes suited for normalization, applied to bladder and colon cancer data sets. Cancer Res. 64, 5245\u20135250. https:\/\/doi.org\/10.1158\/0008-5472.CAN-04-0496 (2004).","journal-title":"Cancer Res."},{"key":"54179_CR15","doi-asserted-by":"publisher","first-page":"164","DOI":"10.3390\/horticulturae8020164","volume":"8","author":"W Xue","year":"2022","unstructured":"Xue, W. et al. Evaluation of reference genes for quantitative PCR in Eustoma grandiflorum under different experimental conditions. Horticulturae 8, 164. https:\/\/doi.org\/10.3390\/horticulturae8020164 (2022).","journal-title":"Horticulturae"},{"key":"54179_CR16","doi-asserted-by":"publisher","first-page":"138","DOI":"10.5482\/HAMO-17-01-0001","volume":"37","author":"C Mannhalter","year":"2017","unstructured":"Mannhalter, C. Neue entwicklungen in der molekularbiologischen diagnostik. Hamostaseologie 37, 138\u2013151. https:\/\/doi.org\/10.5482\/HAMO-17-01-0001 (2017).","journal-title":"Hamostaseologie"},{"key":"54179_CR17","unstructured":"Andrews, S. FastQC: A Quality Control Tool for High Throughput Sequence Data (2010)."},{"key":"54179_CR18","doi-asserted-by":"publisher","first-page":"1509","DOI":"10.1101\/gr.079558.108","volume":"18","author":"JC Marioni","year":"2008","unstructured":"Marioni, J. C., Mason, C. E., Mane, S. M., Stephens, M. & Gilad, Y. RNA-seq: An assessment of technical reproducibility and comparison with gene expression arrays. Genome Res. 18, 1509\u20131517. https:\/\/doi.org\/10.1101\/gr.079558.108 (2008).","journal-title":"Genome Res."},{"key":"54179_CR19","doi-asserted-by":"publisher","first-page":"57","DOI":"10.1038\/nrg2484","volume":"10","author":"Z Wang","year":"2009","unstructured":"Wang, Z., Gerstein, M. & Snyder, M. RNA-Seq: A revolutionary tool for transcriptomics. Nat. Rev. Genet. 10, 57\u201363. https:\/\/doi.org\/10.1038\/nrg2484 (2009).","journal-title":"Nat. Rev. Genet."},{"issue":"Suppl 5","key":"54179_CR20","doi-asserted-by":"publisher","first-page":"v8","DOI":"10.1093\/annonc\/mdv298","volume":"26","author":"E Senkus","year":"2015","unstructured":"Senkus, E. et al. Primary breast cancer: ESMO Clinical Practice Guidelines for diagnosis, treatment and follow-up. Ann. Oncol. 26(Suppl 5), v8-30. https:\/\/doi.org\/10.1093\/annonc\/mdv298 (2015).","journal-title":"Ann. Oncol."},{"key":"54179_CR21","doi-asserted-by":"publisher","first-page":"376","DOI":"10.1016\/j.ncrna.2023.05.001","volume":"8","author":"P Rajendran","year":"2023","unstructured":"Rajendran, P. et al. Salivaomics to decode non-coding RNAs in oral cancer. A narrative review. Non-coding RNA Res. 8, 376\u2013384. https:\/\/doi.org\/10.1016\/j.ncrna.2023.05.001 (2023).","journal-title":"Non-coding RNA Res."},{"key":"54179_CR22","doi-asserted-by":"publisher","first-page":"257","DOI":"10.1038\/nrg.2016.10","volume":"17","author":"SA Byron","year":"2016","unstructured":"Byron, S. A., van Keuren-Jensen, K. R., Engelthaler, D. M., Carpten, J. D. & Craig, D. W. Translating RNA sequencing into clinical diagnostics: Opportunities and challenges. Nat. Rev. Genet. 17, 257\u2013271. https:\/\/doi.org\/10.1038\/nrg.2016.10 (2016).","journal-title":"Nat. Rev. Genet."},{"key":"54179_CR23","first-page":"706","volume":"36","author":"GR Ogden","year":"1992","unstructured":"Ogden, G. R., Cowpe, J. G. & Green, M. Cytobrush and wooden spatula for oral exfoliative cytology. A comparison. Acta Cytol. 36, 706\u2013710 (1992).","journal-title":"Acta Cytol."},{"key":"54179_CR24","doi-asserted-by":"publisher","first-page":"6944","DOI":"10.1038\/s41598-018-25311-0","volume":"8","author":"C Theda","year":"2018","unstructured":"Theda, C. et al. Quantitation of the cellular content of saliva and buccal swab samples. Sci. Rep. 8, 6944. https:\/\/doi.org\/10.1038\/s41598-018-25311-0 (2018).","journal-title":"Sci. Rep."},{"key":"54179_CR25","doi-asserted-by":"publisher","first-page":"501","DOI":"10.1002\/dc.23918","volume":"46","author":"MM Salih","year":"2018","unstructured":"Salih, M. M., El-Esawy, B. H., Abd El Hafez, A. & Abd El-Hafez, A. Cytomorphologic patterns of Pap and PAS-stained oral exfoliative cytology smears in adult Saudi diabetic patients as compared to healthy controls. Diagn. Cytopathol. 46, 501\u2013506. https:\/\/doi.org\/10.1002\/dc.23918 (2018).","journal-title":"Diagn. Cytopathol."},{"key":"54179_CR26","doi-asserted-by":"publisher","DOI":"10.4172\/2157-7145.1000233","author":"D Saini","year":"2014","unstructured":"Saini, D. Usefulness of nucleic acids (DNA\/RNA) from buccal cells isolated from mouthwashes using a modified method. J. For. Res. https:\/\/doi.org\/10.4172\/2157-7145.1000233 (2014).","journal-title":"J. For. Res."},{"key":"54179_CR27","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pone.0229791","volume":"15","author":"R Sullivan","year":"2020","unstructured":"Sullivan, R. et al. An optimised saliva collection method to produce high-yield, high-quality RNA for translational research. PloS one 15, e0229791. https:\/\/doi.org\/10.1371\/journal.pone.0229791 (2020).","journal-title":"PloS one"},{"key":"54179_CR28","doi-asserted-by":"publisher","first-page":"24","DOI":"10.1186\/1755-8794-3-24","volume":"3","author":"DM Kupfer","year":"2010","unstructured":"Kupfer, D. M., White, V. L., Jenkins, M. C. & Burian, D. Examining smoking-induced differential gene expression changes in buccal mucosa. BMC Med. Genomics 3, 24. https:\/\/doi.org\/10.1186\/1755-8794-3-24 (2010).","journal-title":"BMC Med. Genomics"},{"key":"54179_CR29","doi-asserted-by":"publisher","first-page":"116","DOI":"10.1002\/lio2.516","volume":"6","author":"JL Geiger","year":"2021","unstructured":"Geiger, J. L. et al. Clinical trials optimizing investigator and self-collection of buccal cells for RNA yield. Laryngosc. Invest. Otolaryngol. 6, 116\u2013121. https:\/\/doi.org\/10.1002\/lio2.516 (2021).","journal-title":"Laryngosc. Invest. Otolaryngol."},{"key":"54179_CR30","doi-asserted-by":"publisher","first-page":"e237","DOI":"10.1016\/j.rodmex.2018.01.014","volume":"21","author":"MV Madera Anaya","year":"2017","unstructured":"Madera Anaya, M. V. & Su\u00e1rez Causado, A. Evaluation of two RNA extraction methods in children\u2019s saliva. Rev. Odontol. Mex. 21, e237\u2013e243. https:\/\/doi.org\/10.1016\/j.rodmex.2018.01.014 (2017).","journal-title":"Rev. Odontol. Mex."},{"key":"54179_CR31","doi-asserted-by":"publisher","first-page":"9349132","DOI":"10.1155\/2020\/9349132","volume":"2020","author":"T Matsubara","year":"2020","unstructured":"Matsubara, T. et al. DV200 index for assessing RNA integrity in next-generation sequencing. BioMed. Res. Int. 2020, 9349132. https:\/\/doi.org\/10.1155\/2020\/9349132 (2020).","journal-title":"BioMed. Res. Int."},{"key":"54179_CR32","doi-asserted-by":"publisher","first-page":"5647","DOI":"10.1038\/s41467-021-25805-y","volume":"12","author":"E Porcu","year":"2021","unstructured":"Porcu, E. et al. Differentially expressed genes reflect disease-induced rather than disease-causing changes in the transcriptome. Nat. Commun. 12, 5647. https:\/\/doi.org\/10.1038\/s41467-021-25805-y (2021).","journal-title":"Nat. Commun."},{"key":"54179_CR33","doi-asserted-by":"publisher","first-page":"581","DOI":"10.3109\/10520295.2015.1048289","volume":"90","author":"JL Maron","year":"2015","unstructured":"Maron, J. L. & Johnson, K. L. Comparative performance analyses of commercially available products for salivary collection and nucleic acid processing in the newborn. Biotech. Histochem. 90, 581\u2013586. https:\/\/doi.org\/10.3109\/10520295.2015.1048289 (2015).","journal-title":"Biotech. Histochem."},{"key":"54179_CR34","doi-asserted-by":"publisher","first-page":"237","DOI":"10.1016\/j.ymeth.2010.01.010","volume":"50","author":"C Becker","year":"2010","unstructured":"Becker, C., Hammerle-Fickinger, A., Riedmaier, I. & Pfaffl, M. W. mRNA and microRNA quality control for RT-qPCR analysis. Methods (San Diego, Calif.) 50, 237\u2013243. https:\/\/doi.org\/10.1016\/j.ymeth.2010.01.010 (2010).","journal-title":"Methods (San Diego, Calif.)"},{"key":"54179_CR35","doi-asserted-by":"publisher","first-page":"6152","DOI":"10.1073\/pnas.092140899","volume":"99","author":"DK Nam","year":"2002","unstructured":"Nam, D. K. et al. Oligo(dT) primer generates a high frequency of truncated cDNAs through internal poly(A) priming during reverse transcription. Proc. Natl. Acad. Sci. USA 99, 6152\u20136156. https:\/\/doi.org\/10.1073\/pnas.092140899 (2002).","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"54179_CR36","doi-asserted-by":"publisher","first-page":"11147","DOI":"10.1038\/s41598-020-67825-6","volume":"10","author":"P Ostheim","year":"2020","unstructured":"Ostheim, P. et al. Overcoming challenges in human saliva gene expression measurements. Sci. Rep. 10, 11147. https:\/\/doi.org\/10.1038\/s41598-020-67825-6 (2020).","journal-title":"Sci. Rep."},{"key":"54179_CR37","doi-asserted-by":"publisher","first-page":"229","DOI":"10.1016\/j.ymeth.2005.09.003","volume":"37","author":"SD Ginsberg","year":"2005","unstructured":"Ginsberg, S. D. RNA amplification strategies for small sample populations. Methods (San Diego, Calif.) 37, 229\u2013237. https:\/\/doi.org\/10.1016\/j.ymeth.2005.09.003 (2005).","journal-title":"Methods (San Diego, Calif.)"},{"key":"54179_CR38","doi-asserted-by":"publisher","first-page":"509","DOI":"10.1373\/clinchem.2003.026161","volume":"50","author":"A St\u00e5hlberg","year":"2004","unstructured":"St\u00e5hlberg, A., H\u00e5kansson, J., Xian, X., Semb, H. & Kubista, M. Properties of the reverse transcription reaction in mRNA quantification. Clin. Chem. 50, 509\u2013515. https:\/\/doi.org\/10.1373\/clinchem.2003.026161 (2004).","journal-title":"Clin. Chem."},{"key":"54179_CR39","doi-asserted-by":"publisher","first-page":"1","DOI":"10.1093\/bioinformatics\/btx504","volume":"34","author":"KE Kaczor-Urbanowicz","year":"2018","unstructured":"Kaczor-Urbanowicz, K. E. et al. Novel approaches for bioinformatic analysis of salivary RNA sequencing data for development. Bioinformatics (Oxford, England) 34, 1\u20138. https:\/\/doi.org\/10.1093\/bioinformatics\/btx504 (2018).","journal-title":"Bioinformatics (Oxford, England)"},{"key":"54179_CR40","doi-asserted-by":"publisher","first-page":"1081424","DOI":"10.3389\/fgene.2023.1081424","volume":"14","author":"A Kumar","year":"2023","unstructured":"Kumar, A. et al. Microbial contamination and composition of oral samples subjected to clinical whole genome sequencing. Front. Genet. 14, 1081424. https:\/\/doi.org\/10.3389\/fgene.2023.1081424 (2023).","journal-title":"Front. Genet."},{"key":"54179_CR41","doi-asserted-by":"publisher","first-page":"R23","DOI":"10.1186\/gb-2012-13-3-r23","volume":"13","author":"G Giannoukos","year":"2012","unstructured":"Giannoukos, G. et al. Efficient and robust RNA-seq process for cultured bacteria and complex community transcriptomes. Genome Biol. 13, R23. https:\/\/doi.org\/10.1186\/gb-2012-13-3-r23 (2012).","journal-title":"Genome Biol."},{"key":"54179_CR42","doi-asserted-by":"publisher","first-page":"1085","DOI":"10.1373\/clinchem.2017.285072","volume":"64","author":"F Li","year":"2018","unstructured":"Li, F. et al. Characterization of human salivary extracellular RNA by next-generation sequencing. Clin. Chem. 64, 1085\u20131095. https:\/\/doi.org\/10.1373\/clinchem.2017.285072 (2018).","journal-title":"Clin. Chem."},{"key":"54179_CR43","doi-asserted-by":"publisher","first-page":"856","DOI":"10.1016\/j.bbrc.2003.11.177","volume":"313","author":"A Radoni\u0107","year":"2004","unstructured":"Radoni\u0107, A. et al. Guideline to reference gene selection for quantitative real-time PCR. Biochem. Biophys. Res. Commun. 313, 856\u2013862. https:\/\/doi.org\/10.1016\/j.bbrc.2003.11.177 (2004).","journal-title":"Biochem. Biophys. Res. Commun."},{"key":"54179_CR44","doi-asserted-by":"publisher","first-page":"107","DOI":"10.1007\/BF03260053","volume":"8","author":"N Janssens","year":"2004","unstructured":"Janssens, N., Janicot, M., Perera, T. & Bakker, A. Housekeeping genes as internal standards in cancer research. Mol. Diagn. 8, 107\u2013113. https:\/\/doi.org\/10.1007\/BF03260053 (2004).","journal-title":"Mol. Diagn."},{"key":"54179_CR45","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pone.0122515","volume":"10","author":"W de Spiegelaere","year":"2015","unstructured":"de Spiegelaere, W. et al. Reference gene validation for RT-qPCR, a note on different available software packages. PloS one 10, e0122515. https:\/\/doi.org\/10.1371\/journal.pone.0122515 (2015).","journal-title":"PloS one"},{"key":"54179_CR46","doi-asserted-by":"publisher","first-page":"988","DOI":"10.1373\/clinchem.2005.063206","volume":"52","author":"NJ Park","year":"2006","unstructured":"Park, N. J., Li, Y., Yu, T., Brinkman, B. M. N. & Wong, D. T. Characterization of RNA in saliva. Clin. Chem. 52, 988\u2013994. https:\/\/doi.org\/10.1373\/clinchem.2005.063206 (2006).","journal-title":"Clin. Chem."},{"key":"54179_CR47","doi-asserted-by":"publisher","first-page":"64","DOI":"10.1186\/1471-2199-9-64","volume":"9","author":"N Silver","year":"2008","unstructured":"Silver, N. et al. Selection of housekeeping genes for gene expression studies in the adult rat submandibular gland under normal, inflamed, atrophic and regenerative states. BMC Mol. Biol. 9, 64. https:\/\/doi.org\/10.1186\/1471-2199-9-64 (2008).","journal-title":"BMC Mol. Biol."},{"key":"54179_CR48","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pone.0158462","volume":"11","author":"JL Martin","year":"2016","unstructured":"Martin, J. L. Validation of reference genes for oral cancer detection panels in a prospective blinded cohort. PloS one 11, e0158462. https:\/\/doi.org\/10.1371\/journal.pone.0158462 (2016).","journal-title":"PloS one"},{"key":"54179_CR49","doi-asserted-by":"publisher","first-page":"389","DOI":"10.1152\/physiolgenomics.00025.2005","volume":"21","author":"RD Barber","year":"2005","unstructured":"Barber, R. D., Harmer, D. W., Coleman, R. A. & Clark, B. J. GAPDH as a housekeeping gene: Analysis of GAPDH mRNA expression in a panel of 72 human tissues. Physiol. Genomics 21, 389\u2013395. https:\/\/doi.org\/10.1152\/physiolgenomics.00025.2005 (2005).","journal-title":"Physiol. Genomics"},{"key":"54179_CR50","first-page":"155","volume":"15","author":"SA Bustin","year":"2004","unstructured":"Bustin, S. A. & Nolan, T. Pitfalls of quantitative real-time reverse-transcription polymerase chain reaction. J. Biomolx. Tech. JBT 15, 155\u2013166 (2004).","journal-title":"J. Biomolx. Tech. JBT"},{"key":"54179_CR51","doi-asserted-by":"publisher","first-page":"42","DOI":"10.1186\/1741-7007-12-42","volume":"12","author":"I Gallego Romero","year":"2014","unstructured":"Gallego Romero, I., Pai, A. A., Tung, J. & Gilad, Y. RNA-seq: Impact of RNA degradation on transcript quantification. BMC Biol. 12, 42. https:\/\/doi.org\/10.1186\/1741-7007-12-42 (2014).","journal-title":"BMC Biol."},{"key":"54179_CR52","doi-asserted-by":"publisher","first-page":"1601","DOI":"10.1007\/s10529-006-9127-2","volume":"28","author":"S Fleige","year":"2006","unstructured":"Fleige, S. et al. Comparison of relative mRNA quantification models and the impact of RNA integrity in quantitative real-time RT-PCR. Biotechnol. Lett. 28, 1601\u20131613. https:\/\/doi.org\/10.1007\/s10529-006-9127-2 (2006).","journal-title":"Biotechnol. Lett."},{"key":"54179_CR53","doi-asserted-by":"publisher","first-page":"292","DOI":"10.1038\/ng1203-292","volume":"35","author":"H Auer","year":"2003","unstructured":"Auer, H. et al. Chipping away at the chip bias: RNA degradation in microarray analysis. Nat. Genet. 35, 292\u2013293. https:\/\/doi.org\/10.1038\/ng1203-292 (2003).","journal-title":"Nat. Genet."},{"key":"54179_CR54","doi-asserted-by":"publisher","first-page":"3","DOI":"10.1186\/1471-2199-7-3","volume":"7","author":"A Schroeder","year":"2006","unstructured":"Schroeder, A. et al. The RIN: An RNA integrity number for assigning integrity values to RNA measurements. BMC Mol. Biol. 7, 3. https:\/\/doi.org\/10.1186\/1471-2199-7-3 (2006).","journal-title":"BMC Mol. Biol."},{"key":"54179_CR55","doi-asserted-by":"publisher","first-page":"35","DOI":"10.1016\/j.fsigen.2015.03.005","volume":"17","author":"M-H Lin","year":"2015","unstructured":"Lin, M.-H., Jones, D. F. & Fleming, R. Transcriptomic analysis of degraded forensic body fluids. For. Sci. Int. Genet. 17, 35\u201342. https:\/\/doi.org\/10.1016\/j.fsigen.2015.03.005 (2015).","journal-title":"For. Sci. Int. Genet."},{"key":"54179_CR56","doi-asserted-by":"publisher","first-page":"e101","DOI":"10.1182\/blood-2011-03-339705","volume":"118","author":"JW Rowley","year":"2011","unstructured":"Rowley, J. W. et al. Genome-wide RNA-seq analysis of human and mouse platelet transcriptomes. Blood 118, e101\u2013e111. https:\/\/doi.org\/10.1182\/blood-2011-03-339705 (2011).","journal-title":"Blood"},{"key":"54179_CR57","doi-asserted-by":"publisher","DOI":"10.1002\/0471142727.mb0412s94","author":"DC Christodoulou","year":"2011","unstructured":"Christodoulou, D. C., Gorham, J. M., Herman, D. S. & Seidman, J. G. Construction of normalized RNA-seq libraries for next-generation sequencing using the crab duplex-specific nuclease. Curr. Protoc. Mol. Biol. https:\/\/doi.org\/10.1002\/0471142727.mb0412s94 (2011).","journal-title":"Curr. Protoc. Mol. Biol."},{"key":"54179_CR58","doi-asserted-by":"publisher","first-page":"753","DOI":"10.1186\/1756-0500-7-753","volume":"7","author":"EA Chen","year":"2014","unstructured":"Chen, E. A. et al. Effect of RNA integrity on uniquely mapped reads in RNA-Seq. BMC Res. Notes 7, 753. https:\/\/doi.org\/10.1186\/1756-0500-7-753 (2014).","journal-title":"BMC Res. Notes"},{"key":"54179_CR59","doi-asserted-by":"publisher","first-page":"11","DOI":"10.1186\/s12920-020-0664-7","volume":"13","author":"RA Yao","year":"2020","unstructured":"Yao, R. A., Akinrinade, O., Chaix, M. & Mital, S. Quality of whole genome sequencing from blood versus saliva derived DNA in cardiac patients. BMC Med. Genomics 13, 11. https:\/\/doi.org\/10.1186\/s12920-020-0664-7 (2020).","journal-title":"BMC Med. Genomics"},{"key":"54179_CR60","doi-asserted-by":"publisher","first-page":"1437","DOI":"10.1016\/j.ygeno.2019.08.014","volume":"112","author":"O Ibrahim","year":"2020","unstructured":"Ibrahim, O., Sutherland, H. G., Haupt, L. M. & Griffiths, L. R. Saliva as a comparable-quality source of DNA for whole exome sequencing on ion platforms. Genomics 112, 1437\u20131443. https:\/\/doi.org\/10.1016\/j.ygeno.2019.08.014 (2020).","journal-title":"Genomics"}],"container-title":["Scientific Reports"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.nature.com\/articles\/s41598-024-54179-6.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/www.nature.com\/articles\/s41598-024-54179-6","content-type":"text\/html","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/www.nature.com\/articles\/s41598-024-54179-6.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2024,2,19]],"date-time":"2024-02-19T09:10:22Z","timestamp":1708333822000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.nature.com\/articles\/s41598-024-54179-6"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2024,2,19]]},"references-count":60,"journal-issue":{"issue":"1","published-online":{"date-parts":[[2024,12]]}},"alternative-id":["54179"],"URL":"https:\/\/doi.org\/10.1038\/s41598-024-54179-6","relation":{},"ISSN":["2045-2322"],"issn-type":[{"value":"2045-2322","type":"electronic"}],"subject":[],"published":{"date-parts":[[2024,2,19]]},"assertion":[{"value":"18 October 2023","order":1,"name":"received","label":"Received","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"9 February 2024","order":2,"name":"accepted","label":"Accepted","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"19 February 2024","order":3,"name":"first_online","label":"First Online","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"The authors declare no competing interests.","order":1,"name":"Ethics","group":{"name":"EthicsHeading","label":"Competing interests"}}],"article-number":"4061"}}