Fluorescent in situ hybridization in combination with filter cultivation (FISHFC) method for specific detection and enumeration of viable Clostridium perfringens |
| |
Authors: | S Shimizu M Ootsubo Y Kubosawa I Fuchizawa Y Kawai K Yamazaki |
| |
Affiliation: | 1. Division of Marine Life Science, Faculty of Fisheries Sciences, Hokkaido University, Hakodate 041-8611, Japan;2. Research and Development Department, Hokkaido Industrial Technology Center, 379 Kikyo-cho, Hakodate 041-0801, Japan |
| |
Abstract: | To reduce time for enumeration of viable Clostridium perfringens, fluorescence in situ hybridization in combination with filter cultivation (FISHFC) was employed. The method utilized a CLP-180 probe, based on the 16S rRNA region of C. perfringens, and FISHFC fluorescence microscopy to detect C. perfringens, but not organisms from other species. Optimal cultivation requirements for micro-colony formation were TSC medium, anaerobic conditions, 37 °C, and incubation for 6 h. Under these conditions, micro-colony diameters reached 100 μm, a size sufficient for hybridization. Enumeration of C. perfringens using the CLP-180-aided FISHFC method was realized in 9 h as compared to 3–5 days required by the conventional plate count method. Moreover, viable C. perfringens counts of food samples from the two methods were not significantly different. |
| |
Keywords: | Clostridium perfringens Fluorescent in situ hybridization Specific detection |
本文献已被 ScienceDirect 等数据库收录! |
|