Advantages, Disadvantages and Modifications of Conventional ELISA

Hosseini, Samira; Vázquez-Villegas, Patricia; Rito‐Palomares, Marco; Martínez-Chapa, Sergio O.

doi:10.1007/978-981-10-6766-2_5

Cited by 109 publications

(84 citation statements)

References 143 publications

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“…However, these methods require pre-treatment of the sample, expensive tools, qualified personnel and long-time analysis. The enzyme-linked immunosorbent assay (ELISA) is also used to this aim [ 32 , 35 ], but this approach has several disadvantages such as the nonspecific adsorption of substances generating false positives/negatives [ 36 ], the possible matrix effect on enzyme activity [ 37 ] and kit high cost. To overcome these problems, a multitude of sensors for lysozyme have been developed in recent years.…”

Section: Introductionmentioning

confidence: 99%

Molecularly Imprinted Polyscopoletin for the Electrochemical Detection of the Chronic Disease Marker Lysozyme

2020

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Herein we report the electropolymerization of a scopoletin based molecularly imprinted polymer (MIP) for the detection of lysozyme (Lyz), an enzymatic marker of several diseases in mammalian species. Two different approaches have been used for the imprinting of lysozyme based, respectively, on the use of a monomer-template mixture and on the covalent immobilization of the enzyme prior to polymer synthesis. In the latter case, a multi-step protocol has been exploited with preliminary functionalization of gold electrode with amino groups, via 4-aminothiophenol, followed by reaction with glutaraldehyde, to provide a suitable linker for lysozyme. Each step of surface electrode modification has been followed by cyclic voltammetry and electrochemical impedance spectroscopy, which has been also employed to test the electrochemical responses of the developed MIP. The sensors show good selectivity to Lyz and detect the enzyme at concentrations up to 292 mg/L (20 μM), but with different performances, depending on the used imprinting approach. An imprinting factor equal to 7.1 and 2.5 and a limit of detection of 0.9 mg/L (62 nM) and 2.1 mg/L (141 nM) have been estimated for MIPs prepared with and without enzyme immobilization, respectively. Competitive rebinding experiment results show that this sensing material is selective for Lyz determination. Tests were performed using synthetic saliva to evaluate the potential application of the sensors in real matrices for clinical purposes.

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Section: Introductionmentioning

confidence: 99%

Molecularly Imprinted Polyscopoletin for the Electrochemical Detection of the Chronic Disease Marker Lysozyme

2020

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“…However, the detection procedures are more labor-intensive and carry the risk of non-specific binding, which can give false-positive results. Also, ELISA requires centralized laboratory equipment, relatively high sample volume, and has relatively low sensitivity [17,18]. By contrast, nucleic acid hybridizations have the advantage of detecting a specific gene or nucleic-acid sequence, so gene expression is not a prerequisite.…”

Section: Introductionmentioning

confidence: 99%

Development of Loop-Mediated Isothermal Amplification Assay for Rapid Detection of Cucurbit Leaf Crumple Virus

Waliullah

Ling

Cieniewicz

et al. 2020

IJMS

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A loop-mediated isothermal amplification (LAMP) assay was developed for simple, rapid and efficient detection of Cucurbit leaf crumple virus (CuLCrV), one of the most important begomoviruses that infects cucurbits worldwide. A set of six specific primers targeting a total 240 nt sequence regions in the DNA A of CuLCrV were designed and synthesized for detection of CuLCrV from infected leaf tissues using real-time LAMP amplification with the Genie® III system, which was further confirmed by gel electrophoresis and SYBR™ Green I DNA staining for visual observation. The optimum reaction temperature and time were determined, and no cross-reactivity was seen with other begomoviruses. The LAMP assay could amplify CuLCrV from a mixed virus assay. The sensitivity assay demonstrated that the LAMP reaction was more sensitive than conventional PCR, but less sensitive than qPCR. However, it was simpler and faster than the other assays evaluated. The LAMP assay also amplified CuLCrV-infected symptomatic and asymptomatic samples more efficiently than PCR. Successful LAMP amplification was observed in mixed virus-infected field samples. This simple, rapid, and sensitive method has the capacity to detect CuLCrV in samples collected in the field and is therefore suitable for early detection of the disease to reduce the risk of epidemics.

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“…However, ELISA is established on the immunological mechanism & it needed very specific monoclonal & polyclonal sera for sensitive and specific quantification of antigen. Also, it consume long incubation time, mixing & washing steps and it is unable to analyse AFs individually [39].…”

Section: • Energy-assisted Extraction (Eae)mentioning

confidence: 99%

Aflatoxins Occurrence in Spices

Ahmed¹,

Asghar²

2022

Aflatoxins - Occurrence, Detoxification, Determination and Health Risks

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A wide range of spices are used in most dishes as seasoning, colouring, texture developer, palatability or preserving food and beverages worldwide. However, the spices are produced mainly in developing countries where tropical and/or subtropical climate such as high temperature, heavy rainfall and humidity encourage fungal growth leading to increased occurrence of aflatoxins (AFs) in spices. Moreover, the inadequate implementation of good agricultural practice, good manufacturing practice and good hygienic practice in these countries are great alarming situation. AFs are considered as a carcinogenic, mutagenic, teratogenic and immunosuppressive to humans and are classified as hazardous food toxins. This chapter provides the worldwide production and regulations of spices, suitable conditions for the AFs production, worldwide occurrence of AFs, detection techniques and some aspect for the reduction of AFs in spices.

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Advantages, Disadvantages and Modifications of Conventional ELISA

Cited by 109 publications

References 143 publications

Molecularly Imprinted Polyscopoletin for the Electrochemical Detection of the Chronic Disease Marker Lysozyme

Molecularly Imprinted Polyscopoletin for the Electrochemical Detection of the Chronic Disease Marker Lysozyme

Development of Loop-Mediated Isothermal Amplification Assay for Rapid Detection of Cucurbit Leaf Crumple Virus

Aflatoxins Occurrence in Spices

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