Detection of Plasmodium Aldolase Using a Smartphone and Microfluidic Enzyme Linked Immunosorbent Assay

Background. Malaria control efforts are limited in rural areas. A low-cost system to monitor response without the use of electricity is needed. Plasmodium aldolase is a malaria biomarker measured using enzyme linked immunosorbent assay (ELISA) techniques. A three-part system using ELISA was developed consisting of a microfluidic chip, hand crank centrifuge, and a smartphone. Methods. A circular microfluidic chip was fabricated using clear acrylic and a CO2 laser. A series of passive valves released reagents at precise times based upon centrifugal force. Color change was measured via smartphone camera using an application programmed in Java. The microchip was compared to a standard 96-well sandwich ELISA. Results. Results from standard ELISA were compared to microchip at varying concentrations (1–10 ng/mL). Over 15 different microfluidic patterns were tested, and a final prototype of the chip was created. The prototype microchip was compared to standard sandwich ELISA (n=20) using samples of recombinant aldolase. Color readings of standard ELISA and microfluidic microchip showed similar results. Conclusion. A low-cost microfluidic system could detect and follow therapeutic outcomes in rural areas and identify resistant strains.

Download Full-text

Development of Low Cost Enzyme-Linked Immunosorbent Assay Plate Reader

2019 International Symposium on Electronics and Smart Devices (ISESD) ◽

10.1109/isesd.2019.8909658 ◽

2019 ◽

Author(s):

Alfie Rizky Ananda ◽

Donny Danudirdjo ◽

Agung W. Setiawan ◽

Tati Latifah R. Mengko

Keyword(s):

Immunosorbent Assay ◽

Low Cost ◽

Enzyme Linked Immunosorbent Assay ◽

Plate Reader

Download Full-text

An integrated microfluidic system for on-chip enrichment and quantification of circulating extracellular vesicles from whole blood

Lab on a Chip ◽

10.1039/c9lc00624a ◽

2019 ◽

Vol 19 (19) ◽

pp. 3305-3315 ◽

Cited By ~ 4

Author(s):

Yi-Sin Chen ◽

Yu-Dong Ma ◽

Chihchen Chen ◽

Shu-Chu Shiesh ◽

Gwo-Bin Lee

Keyword(s):

Extracellular Vesicles ◽

Immunosorbent Assay ◽

Whole Blood ◽

Magnetic Beads ◽

Extracellular Vesicle ◽

Microfluidic System ◽

Enzyme Linked Immunosorbent Assay ◽

Human Whole Blood ◽

On Chip ◽

Chip Enrichment

An integrated microfluidic system was developed for extracellular vesicle (EV) enrichment and quantification by using anti-CD63-coated magnetic beads and an on-chip enzyme-linked immunosorbent assay in human whole blood.

Download Full-text

A lab in a bento box: an autonomous centrifugal microfluidic system for an enzyme-linked immunosorbent assay

Analytical Methods ◽

10.1039/d0ay01459a ◽

2020 ◽

Vol 12 (40) ◽

pp. 4858-4866

Author(s):

Takaaki Abe ◽

Shunya Okamoto ◽

Akinobu Taniguchi ◽

Michiyasu Fukui ◽

Akinobu Yamaguchi ◽

...

Keyword(s):

Injection Molding ◽

Microfluidic Device ◽

Immunosorbent Assay ◽

Device Driver ◽

Microfluidic System ◽

Enzyme Linked Immunosorbent Assay

In this paper, we report on the demonstration of a portable immunoassay system consisting of a small centrifugal microfluidic device driver (bento box) and a centrifugal microfluidic device made of polypropylene and fabricated by injection molding.

Download Full-text

Aqueous two-phase system antibody confinement enables cost-effective analysis of protein analytes by sandwich enzyme-linked immunosorbent assay with minimal optical crosstalk

The Analyst ◽

10.1039/d0an00699h ◽

2020 ◽

Vol 145 (16) ◽

pp. 5458-5465 ◽

Cited By ~ 1

Author(s):

Maia Kvas ◽

Alyne G. Teixeira ◽

Beatrice Chiang ◽

John P. Frampton

Keyword(s):

Immunosorbent Assay ◽

Low Cost ◽

Cost Effective ◽

Enzyme Linked Immunosorbent Assay ◽

Phase System ◽

Two Phase ◽

Aqueous Two Phase System ◽

Optical Crosstalk ◽

Effective Analysis ◽

Protein Analytes

An aqueous two-phase system was used to reduce reagent volumes and optical crosstalk for a low-cost single sandwich enzyme-linked immunoassay.

Download Full-text

Detection of circulating antigen in bancroftian filariasis by sandwich ELISA using filarial serum IgG

Journal of Helminthology ◽

10.1017/s0022149x00027103 ◽

1984 ◽

Vol 58 (3) ◽

pp. 259-262 ◽

Cited By ~ 30

Author(s):

M. V. R. Reddy ◽

Ashok Malhotra ◽

B. C. Harinath

Keyword(s):

Immunosorbent Assay ◽

Sandwich Elisa ◽

Negative Reaction ◽

Serum Immunoglobulin ◽

Enzyme Linked Immunosorbent Assay ◽

Bancroftian Filariasis ◽

Positive Correlation ◽

Serum Igg ◽

Filarial Antigen ◽

Circulating Antigen

ABSTRACTThe utility of the IgG fraction of human filarial serum immunoglobulin in detecting circulating antigen by sandwich enzyme linked immunosorbent assay (ELISA) was studied. 27 of 33 sera from persons with microfilaracmia, 19 of 30 sera from clinical cases of filariasis, 4 of 30 sera from normal persons from a region endemic for filariasis showed the presence of circulating filarial antigen. All the 20 normal sera from the area where filariasis was not endemic gave negative reaction for filarial antigen. Those sera from persons with microfilaracmia that showed the presence of circulating antigen also showed an apparent positive correlation between the microfilarial density and the antigen titre.

Download Full-text

Portable all-in-one automated microfluidic system (PAMICON) with 3D-printed chip using novel fluid control mechanism

Scientific Reports ◽

10.1038/s41598-021-98655-9 ◽

2021 ◽

Vol 11 (1) ◽

Author(s):

Yushen Zhang ◽

Tsun-Ming Tseng ◽

Ulf Schlichtmann

Keyword(s):

Active Control ◽

Microfluidic Chip ◽

Control Mechanism ◽

State Of The Art ◽

Low Cost ◽

Microfluidic System ◽

Microfluidic Chips ◽

Microfluidic Systems ◽

3D Printed ◽

Pdms Chip

AbstractState-of-the-art microfluidic systems rely on relatively expensive and bulky off-chip infrastructures. The core of a system—the microfluidic chip—requires a clean room and dedicated skills to be fabricated. Thus, state-of-the-art microfluidic systems are barely accessible, especially for the do-it-yourself (DIY) community or enthusiasts. Recent emerging technology—3D-printing—has shown promise to fabricate microfluidic chips more simply, but the resulting chip is mainly hardened and single-layered and can hardly replace the state-of-the-art Polydimethylsiloxane (PDMS) chip. There exists no convenient fluidic control mechanism yet suitable for the hardened single-layered chip, and particularly, the hardened single-layered chip cannot replicate the pneumatic valve—an essential actuator for automatically controlled microfluidics. Instead, 3D-printable non-pneumatic or manually actuated valve designs are reported, but their application is limited. Here, we present a low-cost accessible all-in-one portable microfluidic system, which uses an easy-to-print single-layered 3D-printed microfluidic chip along with a novel active control mechanism for fluids to enable more applications. This active control mechanism is based on air or gas interception and can, e.g., block, direct, and transport fluid. As a demonstration, we show the system can automatically control the fluid in microfluidic chips, which we designed and printed with a consumer-grade 3D-printer. The system is comparably compact and can automatically perform user-programmed experiments. All operations can be done directly on the system with no additional host device required. This work could support the spread of low budget accessible microfluidic systems as portable, usable on-the-go devices and increase the application field of 3D-printed microfluidic devices.

Download Full-text

Enzyme Linked Immunosorbent Assay (ELISA) Technique Guideline

BioScientia Medicina ◽

10.32539/bsm.v5i5.228 ◽

2021 ◽

Vol 5 (5) ◽

pp. 447-453

Author(s):

Rachmat Hidayat ◽

Patricia Wulandari

Keyword(s):

Immunosorbent Assay ◽

Solid Surface ◽

Color Change ◽

Antibody Binding ◽

Enzyme Linked Immunosorbent Assay ◽

Hormone Levels ◽

Elisa Technique ◽

Antibody Complex ◽

Antigen Antibody ◽

Detection Signal

ELISA (Enzyme-linked immunosorbent assay) is a technique used to assess the quantification of peptide, protein, antibody and hormone levels, based on the principle of antigen-antibody binding. In the ELISA technique, antigen immobilization will be carried out on a solid surface, then bound with antibodies to form an antigen-antibody bond complex, where the antigen-antibody complex is bound to the enzyme. The detection signal in the form of a color change will be formed due to the reaction between the enzyme and the substrate.

Download Full-text

Sandwich ELISA for Detection of Pig Meat in Raw Beef Using Antisera to Muscle Soluble Proteins

Journal of Food Protection ◽

10.4315/0362-028x-51.10.790 ◽

1988 ◽

Vol 51 (10) ◽

pp. 790-798 ◽

Cited By ~ 22

Author(s):

ROSARIO MARTÍN ◽

JUAN I. AZCONA ◽

CARMEN CASAS ◽

PABLO E. HERNÁNDEZ ◽

BERNABÉ SANZ

Keyword(s):

Horseradish Peroxidase ◽

Immunosorbent Assay ◽

Peroxidase Activity ◽

Sandwich Elisa ◽

Soluble Proteins ◽

Enzyme Linked Immunosorbent Assay ◽

Specific Antibodies ◽

Double Antibody ◽

Colored Product ◽

Pig Meat

A double-antibody sandwich ELISA (enzyme-linked immunosorbent assay) has been successfully developed for the detection of defined amounts of pig meat (1–50%) in raw beef. Antibodies against pig sarcoplasmic extracts were produced in rabbits. Pig-specific antibodies were affinity purified by removing antibodies which crossreacted with horse, chicken or beef extracts followed by immunoadsorption and elution from a pig-extract column. The ELISA involved capturing antigens in sarcoplasmic extracts with pig specific antibodies immobilized on 96-well plates, detecting bound antigen with pig specific, horseradish peroxidase-labeling antibody, and measuring peroxidase activity by the conversion of a clear substrate to a colored product.

Download Full-text

Evaluation of a Commercial Sandwich Enzyme-Linked Immunosorbent Assay for the Quantification of Beta-Casomorphin 7 in Yogurt Using Solid-Phase Extraction Coupled to Liquid Chromatography-Tandem Mass Spectrometry as the “Gold Standard” Method

Journal of AOAC International ◽

10.5740/jaoacint.17-0155 ◽

2018 ◽

Vol 101 (2) ◽

pp. 515-519 ◽

Cited By ~ 2

Author(s):

Duc Doan Nguyen ◽

Francesco Busetti ◽

Stuart Keith Johnson ◽

Vicky Ann Solah

Keyword(s):

Immunosorbent Assay ◽

Standard Method ◽

Gold Standard ◽

Solid Phase ◽

Sandwich Elisa ◽

Aminopeptidase Activity ◽

Enzyme Linked Immunosorbent Assay ◽

Gold Standard Method ◽

Liquid Chromatography Tandem Mass ◽

Dipeptidyl Aminopeptidase

Abstract This study investigated beta-casomorphin 7 (BCM7) in yogurt by means of LC-tandem MS (MS/MS) and enzyme-linked immunosorbent assay (ELISA) and use LC-MS/MS as the “gold standard” method to evaluate the applicability of a commercial ELISA. The level of BCM7 in milk obtained from ELISA analysis was much lower than that obtained by LC-MS/MS analysis and trended to increase during fermentation and storage of yogurt. Meanwhile, the results obtained from LC-MS/MS showed that BCM7 degraded during stages of yogurt processing, and its degradation may have been caused by X-prolyl dipeptidyl aminopeptidase activity. As a result, the commercial sandwich ELISA kit was not suitable for the quantification of BCM7 in fermented dairy milk.

Download Full-text