为了建立快速的火腿肠中细菌检测和计数方法，本文采用细菌通用探针EUB338作为标记，优化了荧光原位杂交（FISH）技术检测火腿肠中细菌的方法。比较了三种细胞固定方法，结果表明最佳的样品预处理方法为：4 ℃ 4%多聚甲醛固定15 min，46 ℃热固定2 h；最佳的杂交条件：杂交温度为46 ℃，杂交时间为3 h，杂交洗脱液浓度为225 mmol/L；优化的FISH方法应用于火腿肠样品的总菌数的检测，并且将FISH方法对火腿肠样品的细菌计数与传统平板计数方法进行比较，结果显示FISH方法相较于平板计数方法检出数高，而且所需时间大大缩短，操作起来更加方便、简捷。本实验充分体现了FISH快速定量的优势，FISH可以作为食品微生物快速检测的一种有效工具。

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.