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加拿大艾伯塔省西南部腹泻和非腹泻人群粪便中布氏弓形杆菌的比较检测与定量分析

Comparative Detection and Quantification of Arcobacter butzleri in Stools from Diarrheic and Nondiarrheic People in Southwestern Alberta, Canada.

作者信息

Webb Andrew L, Boras Valerie F, Kruczkiewicz Peter, Selinger L Brent, Taboada Eduardo N, Inglis G Douglas

机构信息

Agriculture and Agri-Food Canada, Lethbridge, AB, Canada Department of Biological Sciences, University of Lethbridge, Lethbridge, AB, Canada.

Department of Laboratory Medicine, Chinook Regional Hospital, Lethbridge, AB, Canada.

出版信息

J Clin Microbiol. 2016 Apr;54(4):1082-8. doi: 10.1128/JCM.03202-15. Epub 2016 Feb 10.

Abstract

Arcobacter butzleri has been linked to enteric disease in humans, but its pathogenicity and epidemiology remain poorly understood. The lack of suitable detection methods is a major limitation. Using comparative genome analysis, we developed PCR primers for direct detection and quantification ofA. butzleri DNA in microbiologically complex matrices. These primers, along with existing molecular and culture-based methods, were used to detectA. butzleri and enteric pathogens in stools of diarrheic and nondiarrheic people (n= 1,596) living in southwestern Alberta, Canada, from May to November 2008. In addition, quantitative PCR was used to compare A. butzleridensities in diarrheic and nondiarrheic stools.Arcobacter butzleriwas detected more often by PCR (59.6%) than by isolation methods (0.8%). Comparison by PCR-based detection found no difference in the prevalence ofA. butzleri between diarrheic (56.7%) and nondiarrheic (45.5%) individuals. Rates of detection in diarrheic stools peaked in June (71.1%) and October (68.7%), but there was no statistically significant correlation between the presence ofA. butzleri and patient age, sex, or place of habitation. Densities ofA. butzleriDNA in diarrheic stools (1.6 ± 0.59 log10 copies mg(-1)) were higher (P= 0.007) than in nondiarrheic stools (1.3 ± 0.63 log10copies mg(-1)). Of the 892 diarrheic samples that were positive for A. butzleri, 74.1% were not positive for other bacterial and/or viral pathogens. The current study supports previous work suggesting that A. butzleri pathogenicity is strain specific and/or dependent on other factors, such as the level of host resistance.

摘要

布氏弓形杆菌与人类肠道疾病有关,但其致病性和流行病学仍知之甚少。缺乏合适的检测方法是一个主要限制因素。通过比较基因组分析,我们开发了用于直接检测和定量微生物复杂基质中布氏弓形杆菌DNA的PCR引物。这些引物与现有的基于分子和培养的方法一起,用于检测2008年5月至11月居住在加拿大艾伯塔省西南部的腹泻和非腹泻人群(n = 1596)粪便中的布氏弓形杆菌和肠道病原体。此外,使用定量PCR比较腹泻和非腹泻粪便中布氏弓形杆菌的密度。通过PCR检测到布氏弓形杆菌的频率(59.6%)高于分离方法(0.8%)。基于PCR的检测比较发现,腹泻个体(56.7%)和非腹泻个体(45.5%)中布氏弓形杆菌的患病率没有差异。腹泻粪便中的检测率在6月(71.1%)和10月(68.7%)达到峰值,但布氏弓形杆菌的存在与患者年龄、性别或居住地点之间没有统计学上的显著相关性。腹泻粪便中布氏弓形杆菌DNA的密度(1.6±0.59 log10拷贝mg(-1))高于非腹泻粪便(1.3±0.63 log10拷贝mg(-1))(P = 0.007)。在892份布氏弓形杆菌呈阳性的腹泻样本中,74.1%对其他细菌和/或病毒病原体呈阴性。当前的研究支持了先前的工作,表明布氏弓形杆菌的致病性是菌株特异性的和/或取决于其他因素,如宿主抵抗力水平。

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