Amruta Patil-Joshi1, B E Rangaswamy1, Anjali Apte-Deshpande2. 1. Department of Biotechnology, BIET, Davangere, Karnataka, 577004, India. 2. Central Dogma Pvt Ltd., A4, Gulmohar Residency, Baner Road, Baner, Pune, Maharashtra, 411045, India. anjali@centraldogma.co.in.
Abstract
BACKGROUND: The analysis of the quality of food is important to protect humans from food-borne or food-based illnesses caused by pathogens, such as bacteria, fungi, viruses, and protozoa. Rapid identification of these pathogens is critical to ensure food safety. Various detection and identification strategies exist; however, they are laborious and time consuming and hence the detection takes longer time. The aim of this study was to develop the specific and fast method for the detection of contaminants in milk. RESULTS: In this study, we have developed a simple paper-based PCR method with minimum sample preparation process. The 16S rDNA universal primers were used for the detection of bacterial contaminants. LacZ primers were used for coliform detection which causes serious illness and hence their detection is crucial. ITS region primers were used for fungal detection. The most unique thing about this study is use of Whatman paper no. 1 as sample carrier material. We developed and validated the paper-based PCR method and used it for the detection of microbes and coliforms using milk as a representative sample. CONCLUSION: We evaluated this method for its suitability in the detection of contaminant microbes using different milk samples. The paper-based method could successfully detect contaminants in the milk samples and the results were comparable to the traditional microbial detection method. The traditional microbiological method takes at least 18-20 h for detecting the presence of microbes in any sample but the developed paper-based PCR method can confirm the microbial presence in 2-3 h. This is very promising especially in the testing where sample sterility is crucial.
BACKGROUND: The analysis of the quality of food is important to protect humans from food-borne or food-based illnesses caused by pathogens, such as bacteria, fungi, viruses, and protozoa. Rapid identification of these pathogens is critical to ensure food safety. Various detection and identification strategies exist; however, they are laborious and time consuming and hence the detection takes longer time. The aim of this study was to develop the specific and fast method for the detection of contaminants in milk. RESULTS: In this study, we have developed a simple paper-based PCR method with minimum sample preparation process. The 16S rDNA universal primers were used for the detection of bacterial contaminants. LacZ primers were used for coliform detection which causes serious illness and hence their detection is crucial. ITS region primers were used for fungal detection. The most unique thing about this study is use of Whatman paper no. 1 as sample carrier material. We developed and validated the paper-based PCR method and used it for the detection of microbes and coliforms using milk as a representative sample. CONCLUSION: We evaluated this method for its suitability in the detection of contaminant microbes using different milk samples. The paper-based method could successfully detect contaminants in the milk samples and the results were comparable to the traditional microbial detection method. The traditional microbiological method takes at least 18-20 h for detecting the presence of microbes in any sample but the developed paper-based PCR method can confirm the microbial presence in 2-3 h. This is very promising especially in the testing where sample sterility is crucial.
Entities:
Keywords:
Coliforms; Milk sample; PCR; Rapid test; Whatman paper
Authors: Andrée F Maheux; Dominique K Boudreau; Marc-Antoine Bisson; Vanessa Dion-Dupont; Sébastien Bouchard; Martine Nkuranga; Michel G Bergeron; Manuel J Rodriguez Journal: Appl Environ Microbiol Date: 2014-04-25 Impact factor: 4.792