A Simple Kit of Plate-Trapped Antigen Enzyme-Linked Immunosorbent Assay for Identification of Plant Viruses

Nascimento, Aline Kelly Queiroz do; Lima, J.A.; Barbosa, G. da S.

doi:10.5935/1806-6690.20170025

Cited by 7 publications

(5 citation statements)

References 6 publications

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“…PTA-ELISA was reported to be useful for testing an adequate number of samples for the detection of viruses in a wide range of situations over a short period. A study showed the detection of various plant viruses, such as the Squash mosaic virus (SQMV), Cowpea severe mosaic virus (CPSMV), Cucumber mosaic virus (CMV), Cowpea aphid borne mosaic virus (CABMV), Zucchini yellow mosaic virus (ZYMV), and Papaya lethal yellowing virus (PLYV) in squash, cowpea, and papaya plants [ 61 ]. The detection of Tomato yellow leaf curl Thailand virus, (TYLCTHV) in tomato, pepper, eggplant, okra, and cucurbit plants was analyzed using a TAS-ELISA (triple antibody sandwich enzyme-linked immunosorbent assays) [ 62 ].…”

Section: Methods For Plant Viral Diagnosticmentioning

confidence: 99%

Current Developments and Challenges in Plant Viral Diagnostics: A Systematic Review

Mehetre

Leo

Singh³

et al. 2021

Viruses

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Plant viral diseases are the foremost threat to sustainable agriculture, leading to several billion dollars in losses every year. Many viruses infecting several crops have been described in the literature; however, new infectious viruses are emerging frequently through outbreaks. For the effective treatment and prevention of viral diseases, there is great demand for new techniques that can provide accurate identification on the causative agents. With the advancements in biochemical and molecular biology techniques, several diagnostic methods with improved sensitivity and specificity for the detection of prevalent and/or unknown plant viruses are being continuously developed. Currently, serological and nucleic acid methods are the most widely used for plant viral diagnosis. Nucleic acid-based techniques that amplify target DNA/RNA have been evolved with many variants. However, there is growing interest in developing techniques that can be based in real-time and thus facilitate in-field diagnosis. Next-generation sequencing (NGS)-based innovative methods have shown great potential to detect multiple viruses simultaneously; however, such techniques are in the preliminary stages in plant viral disease diagnostics. This review discusses the recent progress in the use of NGS-based techniques for the detection, diagnosis, and identification of plant viral diseases. New portable devices and technologies that could provide real-time analyses in a relatively short period of time are prime important for in-field diagnostics. Current development and application of such tools and techniques along with their potential limitations in plant virology are likewise discussed in detail.

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Section: Methods For Plant Viral Diagnosticmentioning

confidence: 99%

Current Developments and Challenges in Plant Viral Diagnostics: A Systematic Review

Mehetre

Leo

Singh³

et al. 2021

Viruses

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“…The ELISA indirect test has some advantages: speed of execution, low cost, and possibility of performing a large number of samples, consisting of an excellent seroepidemiological survey tool, which requires screening tests with good sensitivity (SEABRA et al, 2016;NASCIMENTO et al, 2017;KIM et al, 2019). Positive and negative predictive values indicate that the standardized test described here is capable of detecting 53.52% of positives and 58.62% of negatives, offering a good safety margin for a screening test.…”

Section: Discussionmentioning

confidence: 97%

Standardization and validation of an indirect ELISA based on the extraction of proteins from Leptospira spp. as an antigen for the diagnosis of canine leptospirosis

Alves,

Fernandes,

Vieira

et al. 2023

Arq. Inst. Biol.

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Leptospirosis is an important zoonosis. The World Health Organization recommends the use of microscopic soroagglutination in the diagnosis of the disease. However, this is a time-consuming technique that offers a risk of contamination to the performer and requires a trained professional. The indirect enzyme-linked immunosorbent assay (ELISA) test presents less risk, is faster, and has lower cost benefit for high sampling. In view of the advantages of using the test, and the importance of the development of techniques that facilitate and make the diagnosis more efficient, the objective of this study was to standardize an indirect ELISA technique using an immunogenic protein of Leptospira spp. for the diagnosis of canine leptospirosis. The standardized test obtained a cut off of 0.314, 76% of sensitivity, 34% of specificity, 53.52% of positive predictive value, 58.62 of negative predictive value, and 55% of accuracy. The protein extraction technique was efficient for antigen production, and the standardized test can be used in the diagnosis of canine leptospirosis.

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“…Plate Trapped Antigen-ELISA (PTA-ELISA) tests from plant extracts have already been demonstrated as a stable platform for low-cost detection of virus (Mowat and Dawson, 1987;Nascimento et al, 2017). Thus, it could be used to expand access to visceral leishmaniasis diagnosis in low-income countries.…”

Section: Discussionmentioning

confidence: 99%

“…A significant concern toward PTA-ELISA is the stability of this type of test based on non-purified antigens. However, stability of up to 20 months at room temperature has been described for a PTA-ELISA for the detection of plant viruses (Nascimento et al, 2017). Another disadvantage of using total soluble proteins in a PTA-ELISA is that it can lead to a lower sensitivity as the recombinant protein is diluted into the host proteins pool (Garnsey and Cambra, 1993), and the plant compounds could interfere with the reaction.…”

Section: Discussionmentioning

confidence: 99%

Plant Expression of Hydrophobin Fused K39 Antigen for Visceral Leishmaniasis Immunodiagnosis

et al. 2021

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Visceral leishmaniasis is a Neglected Tropical Disease of high mortality caused by the protozoan Leishmania infantum. Its transmission cycle is complex, and it has in the domestic dog its main reservoir. The diagnostic tests currently used rely on prokaryotic systems’ proteins, but their low sensitivity increases the disease’s burden. The plant transient expression of recombinant proteins allows the production of complex antigens. However, this system has limited competitiveness against the bacterial production of purified antigens. Thus, we have shown that the L. infantum K39 antigen’s fusion to a hydrophobin allows its production for diagnostic tests without the need for intensive purification. The sera of naturally infected dogs specifically detect the semi-purified rK39-HFBI protein. The test validation against a panel of 158 clinical samples demonstrates the platform’s viability, resulting in sensitivity and specificity of 90.7 and 97.5%, respectively. Thus, the use of semi-purified antigens fused to hydrophobins can become the standard platform for large-scale antigens production to expand diagnostic tests for other human and veterinary diseases worldwide.

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A Simple Kit of Plate-Trapped Antigen Enzyme-Linked Immunosorbent Assay for Identification of Plant Viruses

Cited by 7 publications

References 6 publications

Current Developments and Challenges in Plant Viral Diagnostics: A Systematic Review

Current Developments and Challenges in Plant Viral Diagnostics: A Systematic Review

Standardization and validation of an indirect ELISA based on the extraction of proteins from Leptospira spp. as an antigen for the diagnosis of canine leptospirosis

Plant Expression of Hydrophobin Fused K39 Antigen for Visceral Leishmaniasis Immunodiagnosis

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