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Home >> Somaclonal Variation, Micropropagation and Tissue Culture >> Protein A Coating - PAC ELISA
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Protein A Coating PAC ELISA - PAC-ELISA. This protein-A coating (PAC) technique involves four steps. The wells are coated with protein-A (lng mrl), high dilution of unprocessed antisera were added, test samples were added and finally antisera (cross absorbed with healthy plant components, to minimize non- specific reaction) were added. This is followed by addition of the enzyme substrate for colour reaction.

The above PAC-ELISA procedure has also been utilized for detection of  mycoplasma like organisms (MLOs) in crude extracts of leaflets and stems of groudnut plants showing witches broom symptoms.

DAS-ELISA method has been used for quantitative estimation of bacterial populations. Rhizobium populations were estimated in many soil samples, when the population was not below 103 cells g-l soil. ELISA was also used for the detection of aflatoxins in groundnut seed samples.

ELISA is currently by far the most widely used test in serological diagnosis. It permits detection of antigens, of widely varying size and morphology. At ICRISAT, near Hyderabad, ELISA has been successfully utilized for study of viruses, MLOs, Rhizobium and aflatoxins.

It can also be used for the detection of mycorrhizal fungi, and gibberellins through the use of monoclonal antibodies (MAbs). At ICRISAT the use of the enzyme penicillase(instead of commonly used costly enzymes like phosphatase, horse radish peroxidase and urease) has also been standardized. (penicillase is cheap and is available in India, while other enzymes need to be imported)

 

  

 
 

 
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