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从人类粪便样本中分离弯曲杆菌。

Isolation of Campylobacter from human stool samples.

作者信息

Salim S M, Mandal J, Parija S C

机构信息

Department of Microbiology, Jawaharlal Institute of Postgraduate Medical Education and Research, Puducherry, India.

出版信息

Indian J Med Microbiol. 2014 Jan-Mar;32(1):35-8. doi: 10.4103/0255-0857.124294.

Abstract

CONTEXT

Campylobacter is an undetected cause of diarrhoea especially under 5 years of age in most of the countries. Isolation of this organism is difficult, expensive and cumbersome.

AIMS

Our objective of this study was to isolate this pathogen from the stool specimens on routinely available blood containing laboratory media using the candle jar for creating the microaerophilic atmosphere in our setup.

SETTINGS AND DESIGNS

A descriptive study.

MATERIALS AND METHODS

A total of 50 stool samples were inoculated onto selective and non-selective media with and without filtration using a 0.45 μm membrane. The inoculated media were simultaneously incubated in microaerophilic conditions using the Anoxomat as well as in candle jars at temperatures 37°C and 42°C. The culture isolates were confirmed by standard phenotypic tests. A simplex polymerase chain reaction (PCR) targeting the 16S ribosomal deoxyribonucleic acid of Campylobacter was performed on the deoxyribonucleic acid (DNA) of the culture isolates as well as on the DNA extracted from the stool filtrates.

STATISTICAL ANALYSIS

Data was expressed as a proportion.

RESULTS

Campylobacter could be isolated in 5 out of 50 stool samples using both the Anoxomat as well as the candle jar. Furthermore, we did not find any difference between the isolation using the selective and blood containing media as well as the different incubation temperatures. All the five were confirmed phenotypically and genotypically to be Campylobacter jejuni. The PCR results corroborated with that of the culture.

CONCLUSIONS

Isolation by culture was as sensitive as that of the PCR.

摘要

背景

弯曲杆菌是腹泻的一个未被发现的病因,在大多数国家,尤其是5岁以下儿童中。分离这种微生物困难、昂贵且繁琐。

目的

本研究的目的是使用烛缸在我们的实验装置中营造微需氧环境,从常规可用的含血实验室培养基上的粪便标本中分离出这种病原体。

设置与设计

一项描述性研究。

材料与方法

总共50份粪便样本接种到选择性和非选择性培养基上,接种时有的经过0.45μm滤膜过滤,有的未经过滤。接种后的培养基同时在微需氧条件下,使用厌氧培养箱以及在烛缸中于37°C和42°C的温度下孵育。培养分离株通过标准表型试验进行确认。对培养分离株的脱氧核糖核酸(DNA)以及从粪便滤液中提取的DNA进行针对弯曲杆菌16S核糖体脱氧核糖核酸的单重聚合酶链反应(PCR)。

统计分析

数据以比例表示。

结果

使用厌氧培养箱和烛缸,在50份粪便样本中的5份中分离出了弯曲杆菌。此外,我们发现在使用选择性培养基和含血培养基以及不同孵育温度进行分离方面没有任何差异。所有5株在表型和基因型上均被确认为空肠弯曲杆菌。PCR结果与培养结果相符。

结论

培养分离与PCR一样敏感。

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