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Development of Indirect Competitive Immuno-Assay Method Using SPR Detection for Rapid and Highly Sensitive Measurement of Salivary Cortisol Levels.

Tahara Y, Huang Z, Kiritoshi T, Onodera T, Toko K - Front Bioeng Biotechnol (2014)

Bottom Line: The surface of an Au chip was modified with PEG6-COOH aromatic dialkanethiol self-assembled monolayers and hydrocortisone 3-(O-carboxymethyl) oxime (hydrocortisone 3-CMO) as a cortisol analog.A detection limit of 38 ppt range with a measurement range of 10 ppt-100 ppb was accomplished without the incubation of a mixing solution consisting of standard cortisol and an anti-cortisol antibody, and the time for quantification of cortisol concentration was 8 min from the sample injection.It was found that the results obtained by the cortisol immuno-assay had a good correlation with those obtained by ELISA assay (R = 0.96).

View Article: PubMed Central - PubMed

Affiliation: Graduate School of Information Science and Electrical Engineering, Kyushu University , Fukuoka , Japan.

ABSTRACT
The monitoring of salivary cortisol as a key biomarker of an individual's stress response has been increasingly focused on. This paper describes the development of a novel cortisol immuno-assay method based on an indirect competitive method using a commercially available surface plasmon resonance instrument. The surface of an Au chip was modified with PEG6-COOH aromatic dialkanethiol self-assembled monolayers and hydrocortisone 3-(O-carboxymethyl) oxime (hydrocortisone 3-CMO) as a cortisol analog. A detection limit of 38 ppt range with a measurement range of 10 ppt-100 ppb was accomplished without the incubation of a mixing solution consisting of standard cortisol and an anti-cortisol antibody, and the time for quantification of cortisol concentration was 8 min from the sample injection. We experimentally compared our immuno-assay with a commercialized salivary cortisol enzyme-linked immunosorbent assay (ELISA) kit using human saliva samples. It was found that the results obtained by the cortisol immuno-assay had a good correlation with those obtained by ELISA assay (R = 0.96). Our findings indicate the potential utility of the cortisol immuno-assay for measurements of human salivary cortisol levels.

No MeSH data available.


Comparison of the measurement values obtained with cortisol biosensor and ELISA using human saliva samples.
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Figure 5: Comparison of the measurement values obtained with cortisol biosensor and ELISA using human saliva samples.

Mentions: Figure 5 shows the results of the comparative experiment using human saliva samples. Linear regression analysis was used to determine the relationship between the measurement values obtained by the biosensor and the ELISA. The correlation coefficient was 0.96 and the regression equation relative the values of cortisol obtained using the SPR (CortisolSPR) and ELISA (CortisolELISA) was CortisolELISA = 0.96 × CortisolSPR + 0.339. The values of slope ±95% confidence interval (CI) and intercept ±95% were 0.96 ± 0.239 and 0.339 ± 0.572, respectively. Thus, it is indicated that the cortisol immuno-assay has good correlation and is capable of performing with similar accuracy to the conventional ELISA. It is necessary to develop the technology for improving sample preparation method in a short time.


Development of Indirect Competitive Immuno-Assay Method Using SPR Detection for Rapid and Highly Sensitive Measurement of Salivary Cortisol Levels.

Tahara Y, Huang Z, Kiritoshi T, Onodera T, Toko K - Front Bioeng Biotechnol (2014)

Comparison of the measurement values obtained with cortisol biosensor and ELISA using human saliva samples.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4126460&req=5

Figure 5: Comparison of the measurement values obtained with cortisol biosensor and ELISA using human saliva samples.
Mentions: Figure 5 shows the results of the comparative experiment using human saliva samples. Linear regression analysis was used to determine the relationship between the measurement values obtained by the biosensor and the ELISA. The correlation coefficient was 0.96 and the regression equation relative the values of cortisol obtained using the SPR (CortisolSPR) and ELISA (CortisolELISA) was CortisolELISA = 0.96 × CortisolSPR + 0.339. The values of slope ±95% confidence interval (CI) and intercept ±95% were 0.96 ± 0.239 and 0.339 ± 0.572, respectively. Thus, it is indicated that the cortisol immuno-assay has good correlation and is capable of performing with similar accuracy to the conventional ELISA. It is necessary to develop the technology for improving sample preparation method in a short time.

Bottom Line: The surface of an Au chip was modified with PEG6-COOH aromatic dialkanethiol self-assembled monolayers and hydrocortisone 3-(O-carboxymethyl) oxime (hydrocortisone 3-CMO) as a cortisol analog.A detection limit of 38 ppt range with a measurement range of 10 ppt-100 ppb was accomplished without the incubation of a mixing solution consisting of standard cortisol and an anti-cortisol antibody, and the time for quantification of cortisol concentration was 8 min from the sample injection.It was found that the results obtained by the cortisol immuno-assay had a good correlation with those obtained by ELISA assay (R = 0.96).

View Article: PubMed Central - PubMed

Affiliation: Graduate School of Information Science and Electrical Engineering, Kyushu University , Fukuoka , Japan.

ABSTRACT
The monitoring of salivary cortisol as a key biomarker of an individual's stress response has been increasingly focused on. This paper describes the development of a novel cortisol immuno-assay method based on an indirect competitive method using a commercially available surface plasmon resonance instrument. The surface of an Au chip was modified with PEG6-COOH aromatic dialkanethiol self-assembled monolayers and hydrocortisone 3-(O-carboxymethyl) oxime (hydrocortisone 3-CMO) as a cortisol analog. A detection limit of 38 ppt range with a measurement range of 10 ppt-100 ppb was accomplished without the incubation of a mixing solution consisting of standard cortisol and an anti-cortisol antibody, and the time for quantification of cortisol concentration was 8 min from the sample injection. We experimentally compared our immuno-assay with a commercialized salivary cortisol enzyme-linked immunosorbent assay (ELISA) kit using human saliva samples. It was found that the results obtained by the cortisol immuno-assay had a good correlation with those obtained by ELISA assay (R = 0.96). Our findings indicate the potential utility of the cortisol immuno-assay for measurements of human salivary cortisol levels.

No MeSH data available.