There are many variables in this assay and caution should be used to have controls in place to have optimal results. ImmunoReagents , Inc.
Product offerings include a wide range of immunochemistry reagents such as purified immunoglobulins , primary antibodies , and secondary antibodies covering a broad spectrum of immunoglobulins from various species. ImmunoReagents also provides custom manufacturing to meet specific customer requirements while adhering to cGMP guidelines and ISO quality systems requirements. The company is located in Raleigh, North Carolina.
Indirect ELISA protocol
In this assay, the protein of interest is usually bound to the ELISA plate, followed by a primary antibody that is directed the protein of interest. Lastly, a labeled secondary antibody is added to the plate.
Pure recombinant ESAT-6 standard product was used in serial dilutions to detect the absorbance and to establish a standard curve from the data; the concentration was on the X axis vs. Tuberculosis TB remains a major public health problem in the twenty-first century.
Indirect ELISA Protocol
The situation is exacerbated by co-infection with human immunodeficiency virus HIV. Yet, timely diagnosis and treatment will have a direct influence on the prognosis. So, new, affordable, sensitive, specific diagnostic assays are required.
Then, we calculated the concentration of each sample from the standard curve. Early diagnosis of TBM is still difficult; besides conventional experimental methods, immunoassays have been recently adopted. Various immunoassays for detecting antigens or antibodies in the serum or CSF of TBM patients have been developed with different sensitivities and specificities. The tuberculin skin test TST is technically simple, but it lacks diagnostic specificity in BCG-vaccinated individuals.
Another widely used diagnostic test is polymerase chain reaction PCR using specific primers as markers for MTB, but this test is not performed correctly in all clinical laboratories and shows variable sensitivity and specificity. Many serological assays have been tried, but nearly all have failed to improve upon the time-honored sputum smear and culture approach.
Cellular imaging system
Currently, it is believed that antigen detection is more sensitive than antibody detection [ 7 ], especially specific MTB antigens such as ESAT-6 and the Ag85 complex. However, the early phase of MTB infection, in patients treated with corticosteroids, or in immune-deficient TBM patients, these tests may provide false negative results when antibody detection is used, and antibody detection cannot distinguish whether it is an acute infection or a latent infection. Therefore, there has been growing interest in antigen detection to diagnose TBM [ 8 ].
Consequently, the possible use of ESAT-6 as a marker of Mycobacterium tuberculosis infection has been proposed in regions with low TB endemicity [ 19 ].
However, in the non-TBM group, four cases were positive. This could be that confirmed, diagnosed TBM patients showed more severe clinical manifestations; so it can be speculated that the level of ESAT-6 in CSF is higher in more severe patients. Therefore, the present assay is appropriate for early diagnosis of TBM with high sensitivity and specificity.
It could be used for developing an immunodiagnostic assay with increased sensitivity and specificity. Open Access This article is distributed under the terms of the Creative Commons Attribution License which permits any use, distribution, and reproduction in any medium, provided the original author s and the source are credited.
ELISA assay development
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- chemicals used by bacteria to perform chemosynthesis.
- Overview of Enzyme-Linked Immunosorbent Assay (ELISA);
- Indirect ELISA protocol | Abcam.
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