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采用巢式聚合酶链反应检测肉类中产肠毒素性产气荚膜梭菌的数量。

Amount of enterotoxigenic Clostridium perfringens in meat detected by nested PCR.

作者信息

Miwa N, Nishina T, Kubo S, Atsumi M, Honda H

机构信息

Shizuoka Prefectural Western Meat Inspection, Center, Kakegawa, Japan.

出版信息

Int J Food Microbiol. 1998 Jul 21;42(3):195-200. doi: 10.1016/s0168-1605(98)00082-8.

Abstract

The incidence and quantity of enterotoxigenic Clostridium perfringens in beef, pork, and chicken meat were determined and compared with that of the total enterotoxigenic and nonenterotoxigenic C. perfringens. The method for the detection and quantification of enterotoxigenic C. perfringens consisted of a combination of the most probable number (MPN) method and a nested polymerase chain reaction after culturing of the sample. The results obtained by this method for inoculated meat samples were significantly correlated with those obtained by the plate count method. When the method was applied to the detection and quantification of enterotoxigenic C. perfringens found in randomly selected meat samples, the organism was found in 2% of the beef pieces (< 10(2) MPN/100 g) and 12% of the chicken pieces (< 10(2)-4.3 x 10(2) MPN/100 g) out of the 50 pieces of each meat tested. No enterotoxigenic C. perfringens was found in pork. Total C. perfringens was found in 16% of the beef (< 10(2)-4.3 x 10(2) MPN/100 g), 10% of the pork (< 10(2) MPN/100 g), and 84% of the chicken (< 10(2)-9.3 x 10(3) MPN/100 g) when 50 pieces of each meat was tested by the conventional MPN method. As shown in the above methods, the majority of cells were not enterotoxigenic cells in the population of C. perfringens. A small number of enterotoxigenic cells of C. perfringens co-existed with a large number of nonenterotoxigenic cells in the same meat sample.

摘要

测定了牛肉、猪肉和鸡肉中产肠毒素性产气荚膜梭菌的发生率和数量,并与总产肠毒素性和非产肠毒素性产气荚膜梭菌进行了比较。产肠毒素性产气荚膜梭菌的检测和定量方法包括最可能数(MPN)法和样品培养后进行的巢式聚合酶链反应。该方法对接种肉样的检测结果与平板计数法的结果显著相关。当该方法应用于随机选择的肉样中产肠毒素性产气荚膜梭菌的检测和定量时,在所检测的每种肉的50块样品中,2%的牛肉块(<10²MPN/100 g)和12%的鸡肉块(<10² - 4.3×10²MPN/100 g)中发现了该菌。猪肉中未发现产肠毒素性产气荚膜梭菌。采用传统MPN法检测每种肉的50块样品时,16%的牛肉(<10² - 4.3×10²MPN/100 g)、10%的猪肉(<10²MPN/100 g)和84%的鸡肉(<10² - 9.3×10³MPN/100 g)中发现了总产气荚膜梭菌。如上述方法所示,产气荚膜梭菌群体中的大多数细胞不是产肠毒素性细胞。在同一块肉样中,少量产肠毒素性产气荚膜梭菌细胞与大量非产肠毒素性细胞共存。

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