Theses and Dissertations
Development of a novel loop-mediated isothermal amplification (LAMP) assay for the rapid detection of Clostridium perfringens
Mississippi State University
Sukumaran, Anuraj Theradiyil
Kiess, Aaron S.
Date of Degree
Graduate Thesis - Campus Access Only
Master of Science (M.S.)
College of Agriculture and Life Sciences
Department of Poultry Science
Clostridium perfringens (C. perfringens) causes diseases such as necrotic enteritis in poultry. Current methods for early detection of C. perfringens, including culture, and molecular techniques (PCR and qPCR), have logistical limitations in detecting pathogens on-site because they require specialized equipment and long sample preparation time. This study describes developing a highly sensitive on-site LAMP assay combined with a simple DNA extraction method to detect C. perfringens. Results showed that the LAMP assay was highly specific and sensitive with the optimal reaction conditions: 63°C for 30 mins with primer concentration of 1.2 mM FIP/BIP and 0.2 mM F3/B3. The HotSHOT method was the most convenient DNA extraction method for the on-site LAMP assay with respect to the processing time, cost, and minimal equipment requirement. Therefore, the HotSHOT DNA extraction method followed by the LAMP assay could serve as a rapid on-site molecular test for C. perfringens in the field.
Chaudhary, Deepa, "Development of a novel loop-mediated isothermal amplification (LAMP) assay for the rapid detection of Clostridium perfringens" (2022). Theses and Dissertations. 5732.