Developing of LAMP-ELISA Assay for rapid, sensitive and cost-effective diagnosis and screening of bacterial infections in wild herbivore population of Tamil Nadu.


Background:

Wildlife disease diagnosis, monitoring, prevention, and control are crucial for wildlife and public health. Wild animals are both target and reservoir hosts of infectious pathogens. Infectious diseases can have a wide range of impacts on wild animals, ranging from subtle but important effects, such as reduced reproduction and life span or increased predation rates, to population declines from lethal disease. Most of these pathogens are zoonotic and capable of cross-spreading across domestic animals and humans. Anthrax, Tuberculosis, and Johne’s disease are the highly prevalent bacterial diseases that are reported in wild herbivores in India (Mondal and Yamage, 2014).

Project Objectives:
Principle

LAMP: Loop-mediated isothermal amplification amplifies the target DNA with high specificity and it’s very rapid. The Bst polymerase used in this technique is highly efficient in strand displacement activity and the rapidness of the amplification increases when using 4 to 6 sets of primers which forms more amplification loops. The LAMP products can be detected using calorimetric, fluorescent detection, and real-time monitoring using a turbidity meter. Here primers are designed specifically to target the partial gene sequence of Bacillus anthracis and Mycobacterium spp. (M. tuberculosis and M. bovis).

ELISA: Enzyme Linked Immuno Sorbent Assay is a much more sensitive and reliable serological technique. The primary antibodies used are specific to Mycobacterium avium paratuberculosiswhich is available in the blood of the infected animals. Like a Sandwich ELISA, primary antibodies capture the antigen of interest and this was detected by a secondary antibody tagged to an enzyme conjugate, changing the color of the substrate when the samples are in the diseased state. ELISA and LAMP can be applied for large-scale screening of the captive animal population.

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