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Clostridium perfringens (C. perfringens), a prolific toxin-producing anaerobe is an important foodborne pathogen with a huge public health concern. Rapid and on-site detection of C. perfringens is of specific importance in developing countries. In the present study, saltatory rolling circle amplification (SRCA) assay was developed for culture-independent, rapid and visual detection of C. perfringens and evaluated in meat with pork as a model. The specificity of the SRCA assay was ascertained by using 62 C. perfringens and 18 non- C. perfringens strains. The analytical sensitivity of the developed SRCA, conventional and real-time PCR assays were 80 fg, 800 fg and 800 fg DNA per tube, respectively. The limit of detection of the SRCA assay was 80 CFU/g of pork in the absence of enrichment and 8 CFU/g after short enrichment of 6 h. The detection limits of 80 CFU/g and 8 CFU/g of pork were attained within 120 min and 8 h, respectively. Real-world or field relevancy of the developed assay was evaluated by screening 82 raw and processed pork samples. As the developed assay is simple, user-friendly, cost-effective and sophisticated-equipment free, it would be more suitable for on-site testing of C. perfringens in foods. To our information, this is the first report to apply SRCA for the detection of C. perfringens. [Display omitted] • A novel SRCA assay was developed for rapid and visual detection of Clostridium perfringens in meat. • The analytical sensitivity of the SRCA assay (80 fg) was 10 fold more than the conventional and real-time PCR. • The detection limit of 80 CFU and 8 CFU per gram of meat was attained within 120 min and 8 h, respectively. • This is the first report to use SRCA for the detection of C. perfringens. [ABSTRACT FROM AUTHOR]

Staphylococcus aureus is one of the most common and leading causes of food poisoning and also responsible for multifarious diseases in humans. Rapid and on-site detection of S. aureus is of specific importance in developing countries. In the present study, a polymerase spiral reaction (PSR) assay was developed for sensitive, rapid and visual detection of S. aureus and validated in meat using pork as a model. The specificity of the PSR assay was ascertained by using 18 S. aureus and 17 non- S.aureus strains. The PSR assay was 100- and 10-fold more sensitive than conventional end-point PCR and real-time PCR, respectively. The limit of detection of the PSR assay was 19.9 × 103 CFU/g of pork without enrichment and 19.9 CFU/g after 6 h enrichment. The detection of 19.9 CFU/g of pork was attained within 8 h. Real-world or field applicability of the developed assay was evaluated by screening 76 raw and processed pork samples. Seeing its rapidity, simplicity, cost-effectiveness, user-friendliness and sophisticated-equipment free nature, this assay has the prospective to turn into the assay of choice for regular detection of S. aureus in foods in less-advanced laboratories. To our knowledge, this is the first report to use PSR for the detection of S. aureus. • A novel PSR assay was developed for rapid and visual detection of Staphylococcus aureus in meat. • The analytical sensitivity of the PSR assay was 100 and 10 fold more than the conventional and real-time PCR, respectively. • The detection of limit of 19.9 × 103 CFU and 19.9 CFU per gram of meat was attained within 2 h and 8 h, respectively. [ABSTRACT FROM AUTHOR]