对比常规培养方法和 FTA 过滤套式 PCR 检测番茄表面的宋内志贺菌和鲍氏志贺菌。

Comparison of conventional culture methods and FTA filtration-nested PCR for the detection of Shigella boydii and Shigella sonnei on tomato surfaces.

机构信息

Department of Food Science and Human Nutrition, University of Florida, 359 FSHN Building, Newell Drive, Gainesville, Florida 32611, USA.

出版信息

J Food Prot. 2005 Aug;68(8):1606-12. doi: 10.4315/0362-028x-68.8.1606.

DOI:10.4315/0362-028x-68.8.1606

PMID:21132967

Abstract

Detection of Shigella boydii UI02 and Shigella sonnei UI05 artificially inoculated onto tomatoes was evaluated using enrichment protocols of the U.S. Food and Drug Administration's Bacteriological Analytical Manual (BAM) and the American Public Health Association's Compendium of Methods for the Microbiological Examination of Food (CMMEF), enrichment in Enterobacteriaceae enrichment (EE) broth supplemented with 1.0 microg/ml novobiocin and incubated at 42 degrees C, and FTA filtration-nested PCR. To assess the effect of natural tomato microflora on recovery, conventional culture enrichments were repeated using rifampin-adapted inocula and enrichment medium supplemented with 50 microg/ml rifampin. The lowest detection levels for S. boydii UI02 were > 5.3 x 10(5) (BAM, CMMEF, and EE broth) and 6.2 CFU per tomato (FTA filtration-nested PCR). For S. sonnei UI05, the lowest detection levels were 1.9 x 10(1) (BAM), 1.5 x 10(3) (CMMEF), 1.1 x 10(1) (EE broth), and 7.4 CFU per tomato (FTA filtration-nested PCR). Natural tomato microflora had a large impact on recovery of S. sonnei UI05 and completely inhibited recovery of S. boydii UI02. EE broth was inhibitory to S. boydii UI02. FTA filtration-nested PCR provided superior detection (P < 0.05) compared with the conventional culture enrichment protocols.

摘要

评估了美国食品和药物管理局（FDA）的《细菌分析手册》（BAM）和美国公共卫生协会的《食品微生物检验手册》（CMMEF）的富集方案、补充有 1.0μg/ml 新生霉素的肠杆菌属富集（EE）肉汤、42°C 孵育以及 FTA 过滤嵌套 PCR 对人工接种于番茄上的宋内志贺菌 UI02 和福氏志贺菌 UI05 的检测效果。为了评估天然番茄菌群对回收的影响，使用适应利福平的接种物和补充有 50μg/ml 利福平的富集培养基重复了常规培养富集。S. boydii UI02 的最低检测水平为>5.3 x 10(5)（BAM、CMMEF 和 EE 肉汤）和每个番茄 6.2 CFU（FTA 过滤嵌套 PCR）。对于 S. sonnei UI05，最低检测水平为 1.9 x 10(1)（BAM）、1.5 x 10(3)（CMMEF）、1.1 x 10(1)（EE 肉汤）和每个番茄 7.4 CFU（FTA 过滤嵌套 PCR）。天然番茄菌群对 S. sonnei UI05 的回收有很大影响，并完全抑制了 S. boydii UI02 的回收。EE 肉汤对 S. boydii UI02 有抑制作用。FTA 过滤嵌套 PCR 与常规培养富集方案相比提供了更好的检测效果（P < 0.05）。