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USE OF ELISA IN DETECTING VARIOUS DISEASES AND ITS FUTURE PERSPECTIVE
TABLE OF CONTENTS
Brief history of ELISA ASSAY.
Year wise developmental data of elisa.
Diseases diagnosed by ELISA.
Old and new approaches of ELISA.
Application of ELISA.
Use of ELISA in nature NANOTECHNOLOGY.
ELISA in future perspective.
This article gives practical description of ELISA test, and their application to hormones, drugs, serum components, auto-immune diseases, infectious diseases and parasitic diseases. It covers the topics such as method of elisa test, its use in detecting various diseases and its future perspective. A full information is given of the ELISA test and its several modifications. A part is devoted to the test's use in parasitology; the present major limitation being the inefficiency in many of the raw antigenic preparations. It also describes the complete procedure of elisa, its types along with its application. This article also throws light on the history of elisa, its current and future status. The work majorly focuses on the broad area of the topic. The presentation in 1971, by Engvoll and perlmann 1, of an enzyme lables in immunoassays impersonated a significant technical advance. Their ELISA demonstrated to become as sensitive as radioimmunoassay but safer to be used. Since then enzyme linked immunosorbent assay has been widely and greatly used in the assay of antibodies and antigens.This review article contains the topics such as history of elisa, method of elisa, types of elisa, its application and role of elisa in future in detecting various diseases.
- The enzyme-linked immunosorbent assay (ELISA) is commonly used in laboratory technique that is used to determine the concentration of an analyte (commomnly antibodies or antigens) in any solution. The basic ELISA, or enzyme linked immuno sorbent assay (ELISA), is differentiated from the other antibody based assays because the separation of a specific
- and a non-specific interactions occurs via successive binding to a solid surface, generally a polystyrene multiwell plate, and because of that quantitative results can be achieved. The steps of the ELISA results in a coloured end product which corresponds to the quantity of analyte present in the original sample or solution.
- ELISA tests are very quick and simple to be carried out, and since these are designed such as to rapidly handle a huge number of samples in parallel, they are the very renouned choice for the evaluation of the various researches and the diagnostic targets.
- ELISA assays uses the antibodies which are covalently linked or bonded ("conjugated") to an enzyme.The antigen is bounded to a plastic well, and the enzyme linked antibody is bounded to the antigen. The unbounded antibody is now rinsed away. The amount of residue enzyme, and hence the amount of the antibody bound, is delineated by adding a substrate which changes the colour when acted upon with the enzyme. The absorbance of the resultant colour change is proportional to the amount of the enzyme that is bounded to the well, therefore to the amount of the antibody, hence to the amount of the antigen. Thus, colour indicates antigen, quantitatively.
- The sensitivity of the ELISA test can be studied by the amplification by enzymatic activity. Each bounded enzyme molecule could generate lots of colored product molecules by the enzyme activity. Prior to enzyme bound antibodies becoming greatly available, the radioactive antibodies were greatly used in the Radio-Immuno-Assays (RIA).
- ELISA assay could be done in many different ways named as "direct", "indirect", "sandwich", and "competition" ELISA.
BREIF HISTORY OF ELISA ASSAY
- Before the development of the ELISA, the one and only option for performing animmunoassaywasradioimmunoassay, a technique that utilizes the radioactivelylabeled antigens or antibodies. In the process of radioimmunoassay, the radioactivity gives the signal that indicates whether any specific antigen or antibody is present in the solution or sample.
- In 1960, Rosalyn Sussman Yalow and Solomon Berson first described the radioimmunoassay process in a scientific paper.
- Since the radioactivity causes a potential health threat, a more safe method has to be soughted. A compatible alternative to the method of radioimmunoassay should replace a nonradioactive signal in the position of the radioactive signal.
- When any specific enzyme reacts with the substrate, a change in color occurs, that is used as a signal. For this process, the signal has to be linked with the presence of the antibody or antigen that is why the enzyme has to be associated to an appropriate antibody.
- Stratis Avrameas and G.B.Pierce developed the process of linking independently.
- Since it is very necessary to remove the unbounded antibody or antigen through washing, the antibody or antigen must fixed to the surface of the container.
- To accomplish this, a technique was put forward by Wide andJerkerin 1966.
YEAR WISE DEVELOPMENTAL DATA OF ELISA
- In 1798 –The very first demonstration of vaccination of the smallpox vaccination was done by EDWARD JENNER.
- In 1890 - Detection of the activity of the antibody against the diphtheria and the tetanus toxins and the uprising of the humoral theory of the immunity by Emil von Behring and Shibasaburo Kitasato.
- In 1900 –Demonstration of the antibody formation theory done by Paul Ehrlich.
- In 1938 –Demonstration of the antigen antibody binding hypothesis given by Sir John Marrack.
- In 1948 – The production of antibodies in plasma B cells.
- Between1959-1962 - The discovery of the structure of the antibodies.
- In 1960 – Description of the Radioimmunoassay method in a scientific paper had done by Sir Rosalyn Yalow and Solomon Berson that was published in 1960.
- In 1966 –Development of a technique that was used to prepare something such as immunosorbent to get the antibodies or antigens fixed to the surface of a well plate was published by Wide and Jerker in the year 1966.
- In1971 –The invention of ELISA by Sir Peter Perlmann and Eva Engvall at the University of Stockholm.
- In 1975 - Production of the monoclonal antibodies for the first time by George Kohler and Cesar.
DISEASES DIAGNOSED BY ELISA
- Diagnosis of 1st, 2nd and 3rd trimesters of pregnancy.
- Diagnosis of hyperthyroidism and hypothyroidism.
- Diagnosis of allergies such as food allergies, insect sting allergies, latex allergies etc.
- Diagnosis of allergic rhinitis commonly known as hay fever.
- Diagnosis of allergic eye diseases.
- Diagnosis of chagas disease caused by bacteria T.cruzi.
- Diagnosis of hyper para thyroidism and hypo para thyroidism.
- Detection of HF( EBOLA HEMORRHAGIC FEVER).
- Diagnosis of human immune deficiency virus.
- Diagnosis of lieshmaniasis.
- Detection of tick borne disease called as( ROCKY MOUNTAIN SPOTTED FEVER) RMSF
- Diagnosis of dengue fever.
- Detection of malaria.
- Diagnosis of WEST NILE VIRUS.
- Detection of influenza.
- Diagnosis of heamolytic anemia.
- Detection of LYME disease that causes cancer.
- Detection of mad cow disease.
- Detection of concentration of serum antibodies.
- Diagnosis of diseases of the foot and mouth.
- Diagnosis of johne’s disease.
OLD AND NEW APPROACHES OF ELISA
- IN DIAGNOSIS OF PARASITIC DISEASES
Presently, the diagnosis and detection of a parasitic infection completely relies on the several methods of laboratory in addition to the clinical symptoms, history of clinic , travelling history, and the geographic location of the patient. The currently used primary tests that are used to diagnose many of the parasitic diseases have made a little change since it has been the development of the microscope. Further, major of the recent tests cannot differentiate between many of the infections are not useful for the following responses to the therapy or for the prognosis. The Recent developments that have occurred in new diagnostic tools, however, have put forward many new avenues for a variety of improvements in the parasitic detection. First of all, a large number of newer assays based on serology which are highly specified and sensitive have emerged, like the Falcon assay screening test ELISA commomly known as the FAST-ELISA, the Dot-ELISA , the rapid antigen detection system known as the RDTS, and the LIPS called as the luciferase immune precipitation system .
- IN DIAGNOSIS OF Q FEVER
The performance of ELISA AND CFT can be evaluated by the diagnosis of Q fever serologically and to demonstrate the role of serology for the demonstration of the shedder status. After performing several experiments, it was observed that CFT was a better tool as compared to ELISA.
APPLICATIONS OF ELISA
- ADVANCEMENTS IN THE ELISA FOR THE FOOD INDUSTRIES
Mostly on a daily basis, new discoveries and developments such as that in the fields of enzymology are rising from the research labs around the world. We know that, enzymes are being extensively used in foods and beverages so that there can be an improvement in the processing efficiency.
- Quantitative Detection of the Surimi by the use of competitive ELISA.
- Detection of Pistachio residues in the processed food by the use of enzyme linked immunosorbent assay.
- USE OF ELISA IN TOXICOLOGY IN THE SCREENING OF DRUGS
The ELISA test kits are useful for forensic toxicology, in the animal toxicology as well as in the screening of drugs and are designed to support for the automated and the semi automated testing equipments.
- USE OF ELISA IN IMMUNOPARASITOLOGY
The use of the Dot-ELISA has extensively in the demonstration of human and the veterinary protozoan the metazoan parasitic diseases that includes the amebiasis, the fascioliasis, the cutaneous and the visceral leishmaniasis, malaria, the schistosomiasis, the toxoplasmosis, the trypanosomiasis and even some of the tick infections. This technique used is rapid, very easy to be performed and to interpret. In addition to it, a slight modification in the Dot-ELISA process allows the demonstration of infection rates of the vectors like ticks and sandflies with the parasites.
Since the ELISA ASSAY performed is used to evaluate either the presence of an antibody or presence of an antigen in a given sample, it becomes a very useful method for the determination of serum antibody concentrations such as HIV or WEST NILE VIRUS etc. Its applications are also used in the food industries in determining the food allergens such as milk, walnuts, peanuts, eggs, almonds etc. apart from these ELISA could be used in toxicology for the screening of certain kinds of drugs.
- The other application of elisa comprises:
- DETECTION OF MYCOBACTERIUM ANTIBODIES IN TUBERCULOSIS
- DETECTION OF HEPATITIS B MARKERS IN SERUM
- DETECTION OF ROTAVIRUS IN FECES
- DETECTION OF HIV ANTIBODIES IN BLOOD
- DETECTION OF ENTEROTOXIN OF E.COLI IN FECES.
USE OF ELISA IN NATURE NANOTECHNOLOGY
- The, currently going strategies for the ultrasensitive determination often requires the sophisticated instruments which may not be available in the laboratories along with the lesser resources. This problem was removed by the introduction of a signal producing mechanism for the process of bio sensing which helps in the detection of a small number of molecules of analyte from the naked eye.
- The enzyme level of the enzyme associated Immuno-sorbent assay known as ELISA checks the growth of the gold nano particles and produces a colored solution along with the specific tonality due to the presence of the analyte. The prostate specified antigen called as PSA and the HIV-1 capsid antigen that is p24 were about to be detected in the complete serum at the ultra low concentration that is 1â€‰×â€‰10−18gm per millilitre.The antigen p24 was also determined with the help of the naked eyes in the serum of HIV-infected persons showing the viral loads that were not able to be detectable with a test based on the gold standard nucleic acid.
ELISA IN FUTURE PERSPECTIVE
Due to the increasing sensitivity and fast performance the solid-phase immunoassays have become immensly popular on regular basis. In order to get freedom from the side effects of radioisotopes and to get rid off the expensive equipments which are required to measure them, have surely made the enzyme-linked immunosorbent assay (ELISA) one of the most fastest growing immunological tests that are being used today. ELISA possesses equal and almost greater sensitivity than the solid-phase immunoradiometric assay.
After many of the research team are committed to research for years not only for theanalysis technology itself, but also with the scientific validation. Normally accepted via ELISA assay technology allows the researchers to identify the antigen-specific T cells at the single cell level, majorly for CD4 or CD8 cell immune response. The number of spots that the elisa assay Cytokine analysis, projected that the spots are produced from a single cell, where as the spots morphological Time Response Analysis, shows that the spot diameter is a direct consequence of the capacity of the cell population. Due to this, the elisa assay has not only become an immunological benchmark technology, but it also allows the researchers for almost natural physiological conditions. The drugs inhibit T cell immunity commonly by two-way mechanism; by the secretion of Cytokine Precursor T-cell populations smaller or by reducing Precursor T cell cytokine production capacity, this technology can provide duplicate information.
This review article is basically a summary of the enzyme linked immunosorbent assay, its technique, its applications and its future aspect. It gives the brief history of elisa, what was the need of its development, its elaborated procedure, its various types etc. Therefore enzyme linked immunosorbent assay(ELISA), is great technique which can diagnose large number of diseases, and it has a huse possibility of improvements which can be made in future to make it more advance and relevant. By making further improvement in the elisa technique many of the lethal diseases which cannot be diagnose earlier in present can be checked and controlled easily in future. Elisa’s hot topics in current are , its use in checking the food allergens in food industries, its use in parasitology, in drug industries and many more. Now a days elisa test is majorly used for diagnosing HIV, in tuberculosis, in hepatitis B, in toxicology , enterotoxins in E.Coli feces etc. The major aim of the review article was to focus on the use of elisa in detecting various diseases.
This review article is helpful in understanding the main procedure of elisa test, its evolution and iys future demand.It will also be helpful in knowing the basic fundamentals and principle of elisa and one can get a brief account of its future. There are great possibilities of more enhancements and improvements under this area.