The impact of saliva collection and processing methods on CRP, IgE, and Myoglobin immunoassays

Clinical and Translational Medicine - Tập 1 Số 1 - 2012
Roslinda Mohamed1, Jennifer L. Campbell1, Justin J. Cooper‐White1,2, Goce Dimeski3,4, Chamindie Punyadeera5,1,2
1The Australian Institute for Bioengineering and Nanotechnology St‐Lucia Australia
2The School of Chemical Engineering and the University of Queensland Queensland Australia
3Chemical Pathology Princess Alexandra Hospital Pathology Queensland Woolloongabba Australia
4School of Medicine Southside Clinical School The University of Queensland Herston Australia
5Saliva Translational Research Group Tissue Engineering and Microfluidic Laboratory the Australian Institute for Bioengineering and Nanotechnology The University of Queensland Old Cooper Road 4072 St. Lucia Queensland Australia

Tóm tắt

AbstractBackgroundOwing to its ease of collection, saliva is potentially the sample of choice in diagnosis. Salivary biomolecules have provided a porthole in surveying a person's health and well‐being. Our study aims were (1) to demonstrate the effects of pre‐analytical steps, collection and pre‐processing techniques on salivary protein detection and (2) to establish an indication of salivary reference intervals for 3 biomolecules of clinical interest.MethodsSaliva samples were collected from participants (n = 25, ages 20–35 years) using the following methods: no stimulation (resting/unstimulated), mechanical, and acid stimulation. The saliva was prepared for analysis by: unprocessed, post standard centrifugation in a container without any additives, and centrifugation using Centrifugal Filter Unit (Amicon® Ultra‐0.5). AlphaLisa® assays were used to measure the levels of C‐Reactive Protein (CRP), Immunoglobin (IgE) and myoglobin in saliva samples.ResultsSaliva flow rates were lowest with the resting/drooling collection method. The lowest total protein concentration was with acid stimulation. Unstimulated and mechanically stimulated collections produced no effect on the CRP and IgE levels while myoglobin levels were highest with the unstimulated collection. Acid stimulation had a negative impact on the measured concentrations of IgE and myoglobin (except for CRP levels).ConclusionMechanical stimulation was the most viable option for collecting saliva without affecting the levels of CRP and myoglobin. The processing methods had an adverse effect on the concentration of total protein as well as on CRP and IgE concentrations.

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Tài liệu tham khảo

10.1067/mpr.2001.113778

Yeh C‐K, 2010, Current development of saliva/oral fluid‐based diagnostics, Texas Dental J, 127, 651

10.1021/pr900675w

SchulzBL Cooper‐WhiteJ PunyadeeraCK: Saliva proteome research: current status and future outlook.Crit Rev Biotechnol2012 doi:https://doi.org/10.3109/07388551.2012.687361.

10.1373/clinchem.2010.153767

Worthman CM, 1990, Sensitive salivary estradiol assay for monitoring ovarian function, Clin Chem, 36, 1769, 10.1093/clinchem/36.10.1769

10.1016/j.jim.2011.07.013

Henson BS, 2010, Collection, Storage, and Processing of Saliva Samples for Downstream Molecular Applications, Methods in Molecular Biology, 666, 21, 10.1007/978-1-60761-820-1_2

Agha Hosseini F, 2012, Relationship of stimulated whole saliva cortisol level with the severity of a feeling of dry mouth in menopausal women, Gerodontology, 29, 43, 10.1111/j.1741-2358.2010.00403.x

10.1111/j.1749-6632.1993.tb18359.x

10.1113/jphysiol.1964.sp007315

Raff H, 2010, Biomarkers: salivary cortisol or cortisone?, Nat Rev Endocrinol, 6, 658, 10.1038/nrendo.2010.192

10.1002/bies.200900013

10.1016/j.jdent.2004.10.009

10.5005/jcdp-9-3-72

10.1038/sj.bdj.4807861

10.1016/j.bbi.2012.02.006

10.1016/j.bbi.2012.01.019

10.1373/clinchem.2009.142240

10.1146/annurev.immunol.21.120601.141103

Stone MJ, 1977, Serum myoglobin level as diagnostic test in patients with acute myocardial infarction, Br Heart J, 39, 375, 10.1136/hrt.39.4.375

10.1016/j.archoralbio.2011.02.007

Ambatipudi KS, 2009, Quantitative analysis of age specific variation in the abundance of human female parotid salivary proteins, J Proteome Res, 8, 5093, 10.1021/pr900478h

10.1161/ATVBAHA.107.157164

10.1208/aapsj0902029

10.1016/j.archoralbio.2007.06.009

10.1021/pr060189z

Marshall T, 1987, Electrophoresis indicates protein loss on centrifugation of whole saliva, Clin Chem, 33, 1263, 10.1093/clinchem/33.7.1263

Levine R, 2004, The scientific basis of oral health education, Community Dent Health, 21, 131

Sreebny LM, 1987, Xerostomia. A neglected symptom, Arch Intern Med, 147, 1333, 10.1001/archinte.1987.00370070145022

Navazesh M, 2008, Measuring salivary flow: Challenges and opportunities, J Am Dent Assoc, 139, 35S, 10.14219/jada.archive.2008.0353

Lipps BV, 2008, Part 3: Elevated levels of the salivary protein immunoglobulin E, nerve growth factor, myoglobin, and insulin in patients with type 2 diabetes or with autoimmunity can be prevented by LT‐10‐treatment, Toxin Reviews, 27, 109, 10.1080/15569540802125530

10.1016/0278-6915(89)90122-1

Stokes JR, 2007, Viscoelasticity of human whole saliva collected after acid and mechanical stimulation, Biorheology, 44, 141

deSouzaFTA et al: Burning mouth syndrome: a therapeutic approach involving mechanical salivary stimulation.Headache: The Journal of Head and Face Pain2011:e180–5.

10.1016/j.forsciint.2005.02.028

10.1177/154405910708600802

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Clinical and Translational Medicine

Tập 1 Số 1

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