We demonstrate: (1) procedures for collection of salivary samples in cognitive impaired older adults by family caregivers in the home setting, (2) procedures for measuring stress activity via salivary cortisol and alpha amylase, and (3) representative profiles. Protocols that allow researchers to study stress-linked processes advance our understanding of biological sensitivity and susceptibility.
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Method Article
Collecting Saliva and Measuring Salivary Cortisol and Alpha-amylase in Frail Community Residing Older Adults via Family Caregivers
In This Article
Summary
Abstract
Salivary measures have emerged in bio-behavioral research that are easy-to-collect, minimally invasive, and relatively inexpensive biologic markers of stress. This article we present the steps for collection and analysis of two salivary assays in research with frail, community residing older adults-salivary cortisol and salivary alpha amylase. The field of salivary bioscience is rapidly advancing and the purpose of this presentation is to provide an update on the developments for investigators interested in integrating these measures into research on aging. Strategies are presented for instructing family caregivers in collecting saliva in the home, and for conducting laboratory analyses of salivary analytes that have demonstrated feasibility, high compliance, and yield quality specimens. The protocol for sample collection includes: (1) consistent use of collection materials; (2) standardized methods that promote adherence and minimize subject burden; and (3) procedures for controlling certain confounding agents. We also provide strategies for laboratory analyses include: (1) saliva handling and processing; (2) salivary cortisol and salivary alpha amylase assay procedures; and (3) analytic considerations.
Introduction
The theoretical perspective guiding lifespan developmental science emphasizes the need to integrate biological, behavioral, social, and contextual factors into the study of health and behavior1. Contemporary theoretical models champion the notion that individual differences in biological sensitivity and susceptibility to contextual factors may moderate risk versus resilience, and can forecast morbidity and mortality in vulnerable populations. Considerable research attention has focused on these issues early in life development1, but only recently have investigators been able to test the core ideas in studies with medically and cognitively frail o....
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Protocol
1. Recruitment
- Obtain appropriate Institutional Review Board approval of collection protocol with special attention to consenting subjects with impaired capacity and the collection of biological specimens9-10.
- Exclude participants with potential for blood leakage into saliva by assessing oral health (i.e. "Did you observe bleeding gums in the past two days?")11.
2. Preparation of Saliva Collection Supplies
- Prior to scheduling home visit to consented participants, gather and assemble necessary supplies: 4 Salimetrics Children's Swabs (SCS) , 4 swab stora....
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Results
The protocol is meant to provide one example of how salivary biomeasures can be collected in the community setting. The procedures are designed to measure the diurnal profile of salivary alpha amylase and salivary cortisol concentrations (Figure 2).
Analytic Strategy
Cortisol results have been shown to be positively skewed. Therefore it is suggested to use log transformation to the data to yield a normal distribution. Several strategies are available t.......
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Discussion
To advance our understanding of the meaning of individual differences in biological sensitivity and susceptibility in frail older adults investigators will need to engage caregivers in the collection of biological specimens. These collection protocols need to be minimally invasive, easy to accomplish, and yield quality specimens. Since one major challenge is ensuring that caregivers collect the samples at the correct time family caregivers designated as saliva collectors should be screened for ability to understand writt.......
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Disclosures
In the interest of full disclosure, DAG is founder and serves as the Chief Scientific and Strategy Advisor at Salimetrics LLC (State College, PA) and SalivaBio LLC (Baltimore, MD). These relationships are managed by the policies of the committee on conflict of interest at the Johns Hopkins University School of Medicine, and Arizona State University's Office of Research Adherence and Integrity.
Acknowledgements
Study funded by NINR, K23NR012017-01A1 Adrenocortical Response and Neuro-psychiatric Symptoms in Alzheimer's Disease.
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Materials
| Name | Company | Catalog Number | Comments |
|---|---|---|---|
| Salimetrics Children Swab (SCS) | Salimetrics | 5001.06 | store at RT 50/pack |
| Swab storage tube (2 ml) | Sarstedt | 51.1534.004 | 100/pack |
| Barcode lables | CISBR | n/a | contact CISBR to order |
| Color coded polypropylene vial caps (for SST) | Sarstedt | (see below) | |
| White | Sarstedt | 65.793.517 | 1,000/pack |
| Blue | Sarstedt | 65.793.513 | 1,000/pack |
| Yellow | Sarstedt | 65.793.514 | 1,000/pack |
| Green | Sarstedt | 65.793.515 | 1,000/pack |
| Orange | Sarstedt | 65.793.516 | 1,000/pack |
| Red | Sarstedt | 65.793.518 | 1,000/pack |
| Black | Sarstedt | 65.793.519 | 1,000/pack |
| Violet | Sarstedt | 65.793.520 | 1,000/pack |
| Mixed Colors | Sarstedt | 65.793.999 | 1,000/pack |
| Salivary Cortisol Enzyme Immuno Assay Kit | Salimetrics | 1.3102 | Diagnostic Kit |
| Salivary Alpha Amylase Enzyme Immuno Assay Kit | Salimetrics | 1.1902 | |
| Nitrile gloves | VWR | (see below) | |
| X-Small | VWR | 82026-423 | 100/box |
| Small | VWR | 82026-424 | 100/box |
| Medium | VWR | 82026-426 | 100/box |
| Large | VWR | 82026-428 | 100/box |
| X-Large | VWR | 82026-43 | 100/box |
| Insulated Shipping Container | ThermoFisher | Varies based on size | Visit VWR to select size and order |
| 4 in Sample Storage Box (7 x 7) | Sarstedt | 95.064.949 | 1 box holds 49 SSTs |
References
- Granger, D. A., Fortunato, C. F., Beltzer, E. K., Virag, M., Bright, M. A., Out, D. Focus on methodology: Salivary bioscience and research on adolescence: An integrated perspective. Journal of Adolescence. 3 (10), 1081-1095 (2012).
- Savla, J. T., Granger, D. A., Roberto, K. A., Davey, A., Blieszner, R., Gwazdauskas, F.
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