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Application of 3D Printing Technology in Increasing the Diagnostic Performance of Enzyme-Linked Immunosorbent Assay (ELISA) for Infectious Diseases.

Singh H, Shimojima M, Shiratori T, An le V, Sugamata M, Yang M - Sensors (Basel) (2015)

Bottom Line: Enzyme-linked Immunosorbent Assay (ELISA)-based diagnosis is the mainstay for measuring antibody response in infectious diseases and to support pathogen identification of potential use in infectious disease outbreaks and clinical care of individual patients.The development of laboratory diagnostics using readily available 3D printing technologies provides a timely opportunity for further expansion of this technology into immunodetection systems.Utilizing available 3D printing platforms, a '3D well' was designed and developed to have an increased surface area compared to those of 96-well plates.

View Article: PubMed Central - PubMed

Affiliation: Department of Intelligent Mechanical Systems, Graduate School of System Design, Tokyo Metropolitan University, 6-6 Asahigaoka, Hino, Tokyo 191-0065, Japan. singhha3@gmail.com.

ABSTRACT
Enzyme-linked Immunosorbent Assay (ELISA)-based diagnosis is the mainstay for measuring antibody response in infectious diseases and to support pathogen identification of potential use in infectious disease outbreaks and clinical care of individual patients. The development of laboratory diagnostics using readily available 3D printing technologies provides a timely opportunity for further expansion of this technology into immunodetection systems. Utilizing available 3D printing platforms, a '3D well' was designed and developed to have an increased surface area compared to those of 96-well plates. The ease and rapidity of the development of the 3D well prototype provided an opportunity for its rapid validation through the diagnostic performance of ELISA in infectious disease without modifying current laboratory practices for ELISA. The improved sensitivity of the 3D well of up to 2.25-fold higher compared to the 96-well ELISA provides a potential for the expansion of this technology towards miniaturization and Lab-On-a-Chip platforms to reduce time, volume of reagents and samples needed for such assays in the laboratory diagnosis of infectious and other diseases including applications in other disciplines.

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Related in: MedlinePlus

Validation by IgG ELISA for rubella virus antibody at 1:100 to 1:6400 dilution of primary antibody (OD405: optical density measured at 405 nm). Points represent mean of OD405 values of 6 tests (n = 6) and error bars represent range of OD405 data obtained. * Asterisk denominates statistical significance (Student’s t-test, p < 0.05).
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sensors-15-16503-f004: Validation by IgG ELISA for rubella virus antibody at 1:100 to 1:6400 dilution of primary antibody (OD405: optical density measured at 405 nm). Points represent mean of OD405 values of 6 tests (n = 6) and error bars represent range of OD405 data obtained. * Asterisk denominates statistical significance (Student’s t-test, p < 0.05).

Mentions: The sensitivity of detection of Rubella virus antibodies in the Rubella virus antibody positive sample was higher using the 3D well following surface modification by chemical etching at all primary antibody dilutions used in the validation (1:100, 1:400, 1:1600 and 1:6400) compared to the 96-well [OD405 (3D well vs. 96 well): 1:100 (3.4 vs. 2.9); 1:400 (2.8 vs. 1.7); 1:1600 (1.6 vs. 0.9 and 1:6400 (0.9 vs. 0.4)]. The sensitivity of detection for the Rubella virus antibody positive sample using the 3D well ELISA was 1.2-fold (1:100), 1.6-fold (1:400), 1.7-fold (1:1600) and 2.25-fold (1:6400) (average: 1.7-fold) higher compared to the 96-well ELISA at each dilution point, respectively. Student’s t-test analysis showed that the sensitivity based on the OD405 values obtained for the Rubella virus antibody positive sample at all dilution points using the 3D well were higher than that of the 96-well (p < 0.05) (Figure 4).


Application of 3D Printing Technology in Increasing the Diagnostic Performance of Enzyme-Linked Immunosorbent Assay (ELISA) for Infectious Diseases.

Singh H, Shimojima M, Shiratori T, An le V, Sugamata M, Yang M - Sensors (Basel) (2015)

Validation by IgG ELISA for rubella virus antibody at 1:100 to 1:6400 dilution of primary antibody (OD405: optical density measured at 405 nm). Points represent mean of OD405 values of 6 tests (n = 6) and error bars represent range of OD405 data obtained. * Asterisk denominates statistical significance (Student’s t-test, p < 0.05).
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Related In: Results  -  Collection

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getmorefigures.php?uid=PMC4541890&req=5

sensors-15-16503-f004: Validation by IgG ELISA for rubella virus antibody at 1:100 to 1:6400 dilution of primary antibody (OD405: optical density measured at 405 nm). Points represent mean of OD405 values of 6 tests (n = 6) and error bars represent range of OD405 data obtained. * Asterisk denominates statistical significance (Student’s t-test, p < 0.05).
Mentions: The sensitivity of detection of Rubella virus antibodies in the Rubella virus antibody positive sample was higher using the 3D well following surface modification by chemical etching at all primary antibody dilutions used in the validation (1:100, 1:400, 1:1600 and 1:6400) compared to the 96-well [OD405 (3D well vs. 96 well): 1:100 (3.4 vs. 2.9); 1:400 (2.8 vs. 1.7); 1:1600 (1.6 vs. 0.9 and 1:6400 (0.9 vs. 0.4)]. The sensitivity of detection for the Rubella virus antibody positive sample using the 3D well ELISA was 1.2-fold (1:100), 1.6-fold (1:400), 1.7-fold (1:1600) and 2.25-fold (1:6400) (average: 1.7-fold) higher compared to the 96-well ELISA at each dilution point, respectively. Student’s t-test analysis showed that the sensitivity based on the OD405 values obtained for the Rubella virus antibody positive sample at all dilution points using the 3D well were higher than that of the 96-well (p < 0.05) (Figure 4).

Bottom Line: Enzyme-linked Immunosorbent Assay (ELISA)-based diagnosis is the mainstay for measuring antibody response in infectious diseases and to support pathogen identification of potential use in infectious disease outbreaks and clinical care of individual patients.The development of laboratory diagnostics using readily available 3D printing technologies provides a timely opportunity for further expansion of this technology into immunodetection systems.Utilizing available 3D printing platforms, a '3D well' was designed and developed to have an increased surface area compared to those of 96-well plates.

View Article: PubMed Central - PubMed

Affiliation: Department of Intelligent Mechanical Systems, Graduate School of System Design, Tokyo Metropolitan University, 6-6 Asahigaoka, Hino, Tokyo 191-0065, Japan. singhha3@gmail.com.

ABSTRACT
Enzyme-linked Immunosorbent Assay (ELISA)-based diagnosis is the mainstay for measuring antibody response in infectious diseases and to support pathogen identification of potential use in infectious disease outbreaks and clinical care of individual patients. The development of laboratory diagnostics using readily available 3D printing technologies provides a timely opportunity for further expansion of this technology into immunodetection systems. Utilizing available 3D printing platforms, a '3D well' was designed and developed to have an increased surface area compared to those of 96-well plates. The ease and rapidity of the development of the 3D well prototype provided an opportunity for its rapid validation through the diagnostic performance of ELISA in infectious disease without modifying current laboratory practices for ELISA. The improved sensitivity of the 3D well of up to 2.25-fold higher compared to the 96-well ELISA provides a potential for the expansion of this technology towards miniaturization and Lab-On-a-Chip platforms to reduce time, volume of reagents and samples needed for such assays in the laboratory diagnosis of infectious and other diseases including applications in other disciplines.

Show MeSH
Related in: MedlinePlus