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幽门弯曲菌的形态学、遗传学和生物化学特性研究,一种从实验鼠肠道中分离出来的新物种。

Morphologic, genetic, and biochemical characterization of Helicobacter magdeburgensis, a novel species isolated from the intestine of laboratory mice.

机构信息

Department of Medical Microbiology, Otto von Guericke University Magdeburg, D-39120 Magdeburg, Germany.

出版信息

Helicobacter. 2010 Oct;15(5):403-15. doi: 10.1111/j.1523-5378.2010.00770.x.

Abstract

BACKGROUND

The presence of enterohepatic Helicobacter species (EHS) is commonly noted in mouse colonies. These infections often remain unrecognized but can cause severe health complications or more subtle host immune perturbations and therefore can confound the results of animal experiments. The aim of this study was to isolate and characterize a putative novel EHS that has previously been detected by PCR screening of specific-pathogen-free mice.

MATERIALS AND METHODS

Biochemical analysis of enzyme activities (API campy), morphologic investigation (Gram-staining and electron microscopy) and genetic analyses (16SrRNA and 23SrRNA analyses, DNA fingerprinting, restriction fragment polymorphisms, and pulsed-field gel electrophoresis) were used to characterize isolated EHS. Genomic DNA fragments were sequenced to develop a species-specific PCR detection assay.

RESULTS

Scanning electron microscopy revealed the presence of spiral-shaped EHS, which varied in length (2.5-6 μm) and contained single monopolar or single bipolar sheathed flagella. The bacteria were grown under anaerobic conditions, preferably on agar plates containing serum or blood. The 16SrRNA, genetic, and biochemical analyses indicated the identification of a novel EHS species, named Helicobacter magdeburgensis. We also examined the genome content using pulsed-field gel electrophoresis. Based on the pattern produced by two restriction enzymes, BamIII and KspI, the genome size was determined to be about 1.7-1.8 Mbp.

CONCLUSION

We isolated and characterized a novel EHS species, H. magdeburgensis, morphologically, biochemically, and genetically. These results are important for future studies on the prevalence and pathophysiologic relevance of such infections. Our PCR assay can be used to detect and discriminate H. magdeburgensis from other Helicobacter species.

摘要

背景

肠肝螺旋杆菌(EHS)在小鼠群体中普遍存在。这些感染通常未被识别,但会导致严重的健康并发症或更微妙的宿主免疫紊乱,因此会干扰动物实验的结果。本研究的目的是分离和鉴定以前通过特定病原体无小鼠 PCR 筛选检测到的假定新型 EHS。

材料与方法

采用酶活性生化分析(API campy)、形态学研究(革兰氏染色和电子显微镜)和遗传分析(16SrRNA 和 23SrRNA 分析、DNA 指纹分析、限制性片段多态性和脉冲场凝胶电泳)来描述分离出的 EHS。对基因组 DNA 片段进行测序以开发一种用于特定物种的 PCR 检测方法。

结果

扫描电子显微镜显示存在螺旋形 EHS,其长度(2.5-6μm)不同,含有单个单极或单个双极鞘鞭毛。细菌在厌氧条件下生长,最好在含有血清或血液的琼脂平板上生长。16SrRNA、遗传和生化分析表明,鉴定出一种新型 EHS 物种,命名为曼海姆螺旋杆菌。我们还使用脉冲场凝胶电泳检查了基因组内容。基于两种限制酶 BamIII 和 KspI 产生的图谱,确定基因组大小约为 1.7-1.8 Mbp。

结论

我们从形态、生化和遗传上分离和鉴定了一种新型 EHS 物种,即 H. magdeburgensis。这些结果对未来研究此类感染的流行情况和病理生理相关性具有重要意义。我们的 PCR 检测方法可用于检测和区分 H. magdeburgensis 与其他螺旋杆菌物种。

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