Cotton fabric as an immobilization matrix for low-cost and quick colorimetric enzyme-linked immunosorbent assay (ELISA)

2014 ◽  
Vol 6 (18) ◽  
pp. 7175-7180 ◽  
Author(s):  
Saeedeh Bagherbaigi ◽  
Emma P. Córcoles ◽  
Dedy H. B. Wicaksono

Cotton-fabric based 2-dimensional colorimetric ELISA which requires a smaller amount of sample and reagents, with quicker assay time.

The Analyst ◽  
2020 ◽  
Vol 145 (16) ◽  
pp. 5458-5465 ◽  
Author(s):  
Maia Kvas ◽  
Alyne G. Teixeira ◽  
Beatrice Chiang ◽  
John P. Frampton

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


2017 ◽  
Vol 2017 ◽  
pp. 1-4
Author(s):  
Nikhil S. Gopal ◽  
Ruben Raychaudhuri

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.


2016 ◽  
Vol 88 (9) ◽  
pp. 4872-4878 ◽  
Author(s):  
Chunxiao Hu ◽  
Ioannis Zeimpekis ◽  
Kai Sun ◽  
Sally Anderson ◽  
Peter Ashburn ◽  
...  

Author(s):  
Martin-Tino Časl ◽  
Anders Grubb

A microtitre plate based enzyme-linked immunosorbent assay for determining the concentration of serum amyloid A (SAA) is described. The method employs easily produced sequence-specific rabbit antibodies and the preferential absorption of SAA to polystyrene, which obviates the use of capture antibodies and allows an assay time of only 3 · 5 h, so that the diagnostic potential of the SAA level as a rapid and reliable marker for inflammation can be fully exploited. The assay has a working concentration range of 0·1–2500 mg/L, which embraces the known biological variation of the SAA concentration. The intra-assay coefficient of variation (CV) for SAA concentrations above 10 mg/L is between 1 · 6 and 3·3% and the inter-assay CV between 3·0 and 4·2%. Recovery of SAA added to serum is from 96 to 102%.


2018 ◽  
Vol 2018 ◽  
pp. 1-9 ◽  
Author(s):  
Bing Wang ◽  
Jincui Gu ◽  
Boyi Chen ◽  
Chengfeng Xu ◽  
Hailing Zheng ◽  
...  

The identification of ancient wool is of great importance in archaeology. Despite lots of meaningful information can be achieved by conventional detection methods, that is, light and electron microscopy, spectroscopy, and chromatography, the efficacy is likely to be limited in the detection of ancient samples with contamination or severe degradation. In this work, an immunoassay was proposed and performed for the identification of ancient wool. First, a specific antibody, which has the benefits of low cost, easy operation, and extensive applicability, was developed directly through immunizing rabbits with complete antigen (keratin). Then, an enzyme-linked immunosorbent assay (ELISA) and a colloidal gold-labelled immunochromatographic strip (ICS) were developed to qualitatively identify the corresponding protein in ancient wool samples unearthed from Kazakhstan and China. The anti-keratin antibody exhibited high sensitivity and specificity for the identification of modern and ancient wool. The limit of detection (LOD) of the ELISA method was 10 ng/mL, and no cross-reactions with other interfering antigens have been noted. It is concluded that the immunoassays are reliable methods for the identification of ancient wool.


2017 ◽  
Vol 6 (5) ◽  
pp. 105
Author(s):  
Nina Difla Muflikhah ◽  
Wayan Tunas Artama

Toxoplasmosis is an infectious disease caused by Toxoplasma gondii, an intracellular protozoan parasite that live inside the cells of the reticulo endothelial and parenchymal cells of human and animals (mammals and birds). Some cases of toxoplasmosis usually have no symptoms, but in any cases caused severe symptoms, such as hydrocephalus, microcephalus, intracranial calcification, retinal damage, brain abscess, mental retardation, lymphadenopathy, and others. Its severe symptoms usually showed a long time after first exposure, except symptoms showed by congenital transmission caused by infected mother. Early diagnosis is important to prevent the illness but methods for toxoplasmosis screening are still too expensive for developing country. Enzyme-linked immunosorbent assay (ELISA) allow the testing of a large number samples within short time frame and based on antibody or antigen detection. This study aimed to know the sensitivity and specificity of recombinat protein GRA1 as antigen using ELISA methods. We tested the sensitivity and spesificity of GRA1 protein as antigen in ELISA methods to diagnose toxoplasmosis and compared with ELISA Kit Commercial. Reliable laboratory testing is important to detect Toxoplasma gondii infection, and focused to improving the low cost and easy-to-use diagnostic instrument. Seventy sera collected and tested using both indirect ELISA, commercial ELISA kit and GRA1 protein coated as antigen. Fourty eight and fifty one samples showed positive IgG antibody result of ELISA-GRA1 and ELISA kit. Negative sample tested by ELISA-GRA1 was 22 samples and 19 sample tested by ELISA Kit. The sensitivity and specificity of GRA1-based on ELISA were 100% and 86.36%, positive prediction value (ppv) was 94.11%. These data indicate that the recombinant protein GRA1 is a highly immunogenic protein in human toxoplasmosis and become a promising marker for the screening of toxoplasmosis.


2017 ◽  
Vol 22 (5) ◽  
pp. 518-528 ◽  
Author(s):  
Guochun Wang ◽  
Champak Das ◽  
Bradley Ledden ◽  
Qian Sun ◽  
Chien Nguyen

Enzyme-linked immunosorbent assay (ELISA) automation for routine operation in a small research environment would be very attractive. A portable fully automated low-cost immunoassay system was designed, developed, and evaluated with several protein analytes. It features disposable capillary columns as the reaction sites and uses real-time calibration for improved accuracy. It reduces the overall assay time to less than 75 min with the ability of easy adaptation of new testing targets. The running cost is extremely low due to the nature of automation, as well as reduced material requirements. Details about system configuration, components selection, disposable fabrication, system assembly, and operation are reported. The performance of the system was initially established with a rabbit immunoglobulin G (IgG) assay, and an example of assay adaptation with an interleukin 6 (IL6) assay is shown. This system is ideal for research use, but could work for broader testing applications with further optimization.


2005 ◽  
Vol 17 (4) ◽  
pp. 350-354 ◽  
Author(s):  
Miguel Salgado ◽  
Elizabeth J. B. Manning ◽  
Michael T. Collins

Antibody detection–based tests for paratuberculosis offer speed and economy, 2 diagnostic test attributes important to animal industries with narrow profit margins. Application of such tests to individual milk samples instead of serum samples can further improve testing efficiency and decrease testing cost. Accuracy of a commercial bovine paratuberculosis enzyme-linked immunosorbent assay (ELISA) adapted for use on goat serum and milk samples was determined. Fecal, blood, and milk samples were collected from 159 goats belonging to 2 Wisconsin goat herds with a prior history of paratuberculosis and 1 herd of 50 goats from a paratuberculosis-free Wisconsin herd. Fecal samples were cultured using the modified BACTEC 12B media. Sera were tested according to the manufacturer's instructions for bovine samples. Milk samples were centrifuged and mixed with the ELISA kit's Mycobacterium phlei–containing diluent at a ratio of 1:2. Using fecal culture as the “gold standard,” the sensitivity of the ELISA on goat serum was 64% and the sensitivity of the ELISA on goat milk was 48%. The milk ELISA had higher agreement with fecal culture results (kappa = 0.525) than the serum ELISA (kappa = 0.425). ELISA specificity was 100% on both serum and milk. Regression analysis also showed good correlation between serum and milk S/P values ( r2 = 0.67). Although less sensitive, the ELISA on goat milk samples appears to offer a useful, low-cost alternative for detection of goats with paratuberculosis that have progressed to the stage of shedding M. paratuberculosis in their feces.


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