Salahaddin University – College of Agriculture

Engineering Sciences Stage 4

ELISA methods for plant virus detection

and identification

by:
Asma Munthir Kakahama

Supervised by :
Dr. Khalid Mahmmod Ahmed
Outline:
 Introduction
 Aims
 ELISA
 Typer of elisa
 Direct ELISA
 Indirect ELISA
 Triple Antibody Sandwich (TAS- ELISA)
 Protein A-Sandwich (PAS- ELISA)
 Conclusion
 Introduction
 The diseases caused by virus are generally difficult to control and several
of them cause serious problems for the crop productions.

 Virus infections generally results in plant decline and consequent

reduction in the plants and orchards longevity, with reduction of quality
and quantity of production.

 An adequate diagnosis procedure with a correct virus identification and

determination of its distribution in the field are essential for establishing
efficient control measures.

 Among several serological methods developed and used successfully for

plant virus identification the enzymelinked immunosorbent assay (ELISA)
has been very useful and popular since it was introduced to plant virology
by Clark and Adams (1977).
 Aims
The aims of Project:
To know ELESA test in diagnosing plant virus.

Know the types of Elisa its modification.

 ELISA
 Enzyme-linked immunosorbent assay (ELISA) 
 The ELISA has been used as a diagnostic tool in medicine and
plant pathology, as well as a qualitycontrol check in various
industries. 
 ELISA is a popular format of a "wet-lab" type analytic biochemistry
assay that is used to detect the presence of a substance, usually
an antigen, in a liquid sample or wet sample.
 Types of elisa:
Four types 
Indirect elisa 
Direct elisa 

Competitive elisa 

Multiple and portable elisa

 Direct ELISA
 The direct ELISA , also called double antibody sandwich (DAS-
ELISA), is highly strain-specific and requires each detecting
antibody to be conjugated to an enzyme.
 Typically, the enzyme is alkaline phosphatase.
 Indirect ELISA
 It utilizes a primary un-labeled antibody in conjunction with a
labeled secondary antibody.
 Secondary antibody has specificity for primary antibody.
 The indirect ELISA also requires the use of a universal IgG
enzyme conjugate which can be used with the antibodies of all
virus species.
 Triple Antibody Sandwich (TAS- ELISA)
 Antigens like Tumor markers, hormones, serum proteins may be
determined.
 Antigens in the sample bind with the capture antibody & become
immobilized.
 The antibody of the enzyme conjugate bind with the immobilized
antigen to form a sandwich of Ab-Ag-Ab/ enzyme bound to
microwell.
Protein A-Sandwich (PAS- ELISA)
 Conclusion
 Economic loss has been estimated more than several billion dollars
per year worldwide due to plant viral diseases and there is no
commercialized chemical to manage those .

 Symptomatic diagnosis is still useful but often has erroneous results,

because of confusion associated with high variable symptoms by
interactions between host and virus or by abiotic stresses.

 The accurate and rapid identification of the organisms that cause

plant disease is essential for effective disease control. The methods
based on serological principle and have been used for virus
diagnosis. ELISA associated with serology was initiated and adapted
as a diagnostic tool worldwide since it is easy to use with durability.
THANKS