Developing a qPCR analysis protocol for lactic acid bacteria genera causing meat spoilage

Lactic acid bacteria (LAB) species are widely associated with fresh as well as cooked meat products and are the prevailing spoilage organism in packed meat products. Species composition and metabolic activities of such LAB spoilage communities are determined by the nature of the meat product, storage conditions, and interspecies interactions. Traditional microbiological methods for detecting spoilage through total bacterial counts can be time-consuming and prone to interference. In recent years, quantitative polymerase chain reaction (qPCR) has emerged as a rapid and more accurate tool for detecting and quantifying LAB. In this thesis, we present a qPCR protocol for detecting LAB genera responsible for meat spoilage by designing primers on the genus level targeting unique genes of LAB genera commonly associated with meat spoilage. A pure culture standard curve was used to analyze meat samples and calculated the CFU/g. The total bacterial count obtained by the plate count method was compared with qPCR analysis of the same sample and found that the qPCR protocol can accurately detect and quantify LAB genera causing meat spoilage. The developed qPCR protocol provides a rapid and reliable tool for detecting and quantifying LAB genera, which can help prevent spoilage and ensure the safety of meat products. This method can also be used in the food industry for a quick quality check of products and ingredients.