Optimization of Fluorescence in Situ Hybridization for the Detection of Bacteria in Sausage Samples
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Abstract:
In order to develop a method that rapidly detect and enumerate the bacteria in sausage samples. The Fluorescence in situ hybridization (FISH) technique with EUB338 probe was established and optimized. Three different cell-fixing methods were contrasted and results showed that the optimal conditions for sample preparation were 4% paraformaldehyde, fixed time 15 min, at 4℃and heat fixed time 2 h at 46 ℃. The optimal condition for hybridization was 3 h hybridized at 46℃ with 225 mmol/L NaCl concentration in washing buffer. Compared to the Plate count method, the optimized FISH method was more sensitive and quicker while used to enumerate the bacteria in sausage samples. The FISH method can be used as an effective and convenient tool for the detection and enumeration of bacteria in food samples.