Abstract

Review Article

Three modern serological methods to detect plant viruses

Mahmoud Hamdy Abd El-Aziz*

Published: 10 October, 2019 | Volume 3 - Issue 3 | Pages: 101-106

The use of enzyme linked immunosorbent assay (ELISA) for the detection of plant viruses is well documented. It proved to be a very valuable detection tools for the plant viruses. The efficiency of the ELISA technique was for practical purpose independent of the ratio of antibodies to antigen. This avoids the necessity of making specific enzyme conjugates for each antigen to be tested and eliminates the extreme specificity, thus allowing for quantitative evaluation of strain relationships. The advantages of indirect ELISA are sample. It needs only to be macerated and added to the plate. The crude antiserum could be used, although it should be cross absorbed before to prevent spurious host reaction. Single commercially available second antibody conjugate is utilized, thus eliminating the problems of preparing and storing many different conjugated antisera. Blotting technique has become widely used for specific identification of nucleic acid and proteins. This dot assay was modified to detect protein by spotting the antigen on a nitrocellulose membrane and incubating the membrane in test antibody followed by incubation in peroxidase-conjugated second antibody to the first antibody, and by development in 4-chloro-1-naphthol. The above procedure termed dot blot immunobinding assay (DBIA). The technique of tissue blotting on nitrocellulose membrane was described for detection of plant viruses in infected plants. Tissue blots were made by pressing with a firm and gentile force, the freshly cut tissue surface on nitrocellulose membranes. The possibility of using both sides of the nitrocellulose membrane (NCM) by tissue blot immuno assay (TBIA) for the detection plant viruses. In an effort to reduce the cost of virus assays, different types of regular paper were evaluated as possible replacements for the commonly used nitrocellulose membrane (NCM) as the solid phase in the tissue-blot immunoassay (TBIA) were used. Comparisons between different serological methods were demonstrated by many investigators Dot immunobinding was eight times more sensitive for detection of PVX and four times more sensitive for detection of PVS and PVY than DAS-ELISA.

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Keywords:

ELISA; TBIA; DBIA; Antibodies; NCM

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