Theses and Dissertations

Issuing Body

Mississippi State University

Advisor

Zhang, Li

Committee Member

Sukumaran, Anuraj Theradiyil

Committee Member

Cheng, Wen-Hsing

Committee Member

Kiess, Aaron S.

Date of Degree

12-9-2022

Document Type

Graduate Thesis - Open Access

Major

Poultry Science

Degree Name

Master of Science (M.S.)

College

College of Agriculture and Life Sciences

Department

Department of Poultry Science

Abstract

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.

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