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一种增强型技术,结合预富集和被动过滤,提高了从水样和动物粪便样本中分离空肠弯曲菌和结肠弯曲菌的效率。

An enhanced technique combining pre-enrichment and passive filtration increases the isolation efficiency of Campylobacter jejuni and Campylobacter coli from water and animal fecal samples.

机构信息

Laboratory for Foodborne Zoonoses, Public Health Agency of Canada, Lethbridge, Alberta, Canada.

出版信息

J Microbiol Methods. 2012 Dec;91(3):506-13. doi: 10.1016/j.mimet.2012.09.005. Epub 2012 Sep 14.

Abstract

Improved isolation techniques from environmental water and animal samples are vital to understanding Campylobacter epidemiology. In this study, the efficiency of selective enrichment in Bolton Broth (BB) followed by plating on charcoal cefoperazone deoxycholate agar (CCDA) (conventional method) was compared with an approach combining BB enrichment and passive filtration (membrane method) adapted from a method previously developed for testing of broiler meat, in the isolation of thermophilic campylobacters from surface water and animal fecal samples. The conventional method led to recoveries of Campylobacter from 36.7% of the water samples and 78.0% of the fecal samples and similar numbers, 38.3% and 76.0%, respectively, were obtained with the membrane method. To investigate the genetic diversity of Campylobacter jejuni and Campylobacter coli obtained by these two methods, isolates were analyzed using Comparative Genomic Fingerprinting, a high-resolution subtyping technique. The conventional and membrane methods yielded similar numbers of Campylobacter subtypes from water (25 and 28, respectively) and fecal (15 and 17, respectively) samples. Although there was no significant difference in recovery rates between the conventional and membrane methods, a significant improvement in isolation efficiency was obtained by using the membrane method, with a false-positive rate of 1.6% compared with 30.7% obtained using the conventional method. In conclusion, although the two methods are comparable in sensitivity, the membrane method had higher specificity, making it a cost-effective procedure for the enhanced isolation of C. jejuni and C. coli from water and animal fecal samples.

摘要

改进从环境水样和动物样本中分离的技术对于了解弯曲杆菌病的流行病学至关重要。在本研究中,比较了在 Bolton 肉汤(BB)中进行选择性富集,然后在炭头孢哌酮去氧胆酸盐琼脂(CCDA)上平板(常规方法)与结合 BB 富集和被动过滤(膜方法)的方法,从地表水和动物粪便样本中分离嗜热性弯曲杆菌。常规方法从 36.7%的水样和 78.0%的粪便样本中回收了弯曲杆菌,而膜方法分别获得了 38.3%和 76.0%的回收率。为了研究通过这两种方法获得的空肠弯曲菌和大肠弯曲菌的遗传多样性,使用比较基因组指纹图谱(一种高分辨率的分型技术)对分离株进行了分析。常规方法和膜方法从水样(分别为 25 个和 28 个)和粪便(分别为 15 个和 17 个)样本中获得了相似数量的弯曲杆菌亚型。尽管常规方法和膜方法的回收率之间没有显著差异,但使用膜方法获得了更高的分离效率,假阳性率为 1.6%,而常规方法的假阳性率为 30.7%。总之,尽管这两种方法在灵敏度上具有可比性,但膜方法具有更高的特异性,使其成为从水样和动物粪便样本中增强空肠弯曲菌和大肠弯曲菌分离的一种具有成本效益的方法。

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