The use of bioluminescent enzyme bioassay for the analysis of human saliva: Advantages and disadvantages

• Published in: Luminescence (Chichester, England) England), 2024-05, Vol.39 (5), p.e4776-n/a
• Main Authors: Kolenchukova, Oksana A., Dedora, Anastasia O., Stepanova, Lyudmila V., Kravchuk, Vlada U., Kratasyuk, Valentina A.
• Format: Article
• Language: English
• Subjects: Bio-assays; Bioassays; Biological Assay; Biological effects; Bioluminescence; biomarkers; Cortisol; Enzyme immunoassay; Enzymes; Hormones; Humans; Hydrocortisone - analysis; Hydrocortisone - metabolism; Immunoassay; Immunoassays; Immunoglobulin A; Immunoglobulin A, Secretory - analysis; Immunoglobulin A, Secretory - metabolism; Luciferases - chemistry; Luciferases - metabolism; Luminescence; Luminescent Measurements - methods; NADH:FMN oxidoreductase + reductase; Nicotinamide adenine dinucleotide; Oxidoreductase; Oxidoreductases; Pharyngitis; Physiological effects; Saliva; Saliva - chemistry; Saliva - enzymology; Spectrophotometry; Training
• ISSN: 1522-7235; 1522-7243
• DOI: 10.1002/bio.4776

The purpose of the work was to find optimal conditions for bioluminescent enzymatic analysis of saliva (based on the use of NADH:FMN oxidoreductase + luciferase) and then to determine the biological effect of using bioluminescence assay of saliva to study the physiological state of the body under normal and pathological conditions. The saliva of snowboarders and students were studied in the “rest–training” model. The saliva of patients diagnosed with acute pharyngitis was examined in the “sick–healthy” model. Bioluminescence assay was performed with a lyophilized and immobilized bi‐enzyme system using cuvette, plate, and portable luminometers. The concentrations of secretory immunoglobulin A (sIgA) and cortisol were determined by enzyme immunoassay, and the total protein content was measured by spectrophotometric method. The activity of the bioluminescent system enzymes increased as the amount and volume of saliva in the sample was decreased. The cuvette and plate luminometers were sensitive to changes in the luminescence intensity in saliva assay. Luminescence intensity correlated with the concentrations of sIgA and cortisol. The integrated bioluminescent index for saliva was reduced in the “rest–training” model and increased in the “sick–healthy” model. Thus, the non‐invasive bioluminescent saliva analysis may be a promising tool for assessing the health of the population. The work is devoted to the study of the mechanisms of bioluminescent testing of saliva, depending on the type of luminometer, the amount of saliva, and the effect of its metabolites on bioluminescent glow, as well as the possibility of testing saliva by bioluminescent method during exercise and in the presence of pathology.