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恒河猴幽门螺杆菌分离株的特征鉴定及初步鉴定

Characterization and presumptive identification of Helicobacter pylori isolates from rhesus monkeys.

作者信息

Drazek E S, Dubois A, Holmes R K

机构信息

Department of Medicine, Uniformed Services University of the Health Sciences, Bethesda, Maryland 20814.

出版信息

J Clin Microbiol. 1994 Jul;32(7):1799-804. doi: 10.1128/jcm.32.7.1799-1804.1994.

Abstract

We characterized 38 Helicobacter isolates, including 22 from gastric biopsy samples obtained from 14 rhesus monkeys and single isolates from 16 monkeys in a different colony. Biochemical profiles of these isolates were nearly identical to that of Helicobacter pylori ATCC 43504. Restriction fragment length polymorphism (RFLP) analysis indicated that each infected monkey harbored one to four strains. The 17 RFLP types found among these 22 isolates differed from all seven RFLPs found among the other 16 isolates. Thus, monkeys within a given colony are more likely to be infected by Helicobacter isolates with the same or a similar RFLP than are monkeys from different colonies. A 16S rRNA gene was amplified by PCR and cloned from the Helicobacter isolate from rhesus monkey 85D08. Ribotyping with this probe demonstrated less diversity among isolates from rhesus monkeys than was reported among isolates of H. pylori from humans, as did RFLP analysis of a PCR fragment of the ureA-ureB gene cluster. The DNA sequence of the cloned 16S rRNA gene was determined and compared with sequences reported for H. pylori and other Helicobacter species. Our analysis of 127 nucleotides (corresponding with residues 1240 to 1366 of the Escherichia coli 16S rRNA gene) indicated that the Helicobacter isolate from monkey 85D08 was 99.2 to 100% homologous to isolates of H. pylori from humans but only 83.5 to 96.9% homologous with other Helicobacter species in this region of the 16S rRNA gene. These data provide strong support for the presumptive identification of these isolates as H. pylori.

摘要

我们对38株幽门螺杆菌分离株进行了特征分析,其中22株来自14只恒河猴的胃活检样本,另外16株来自不同猴群的单株分离物。这些分离株的生化特征与幽门螺杆菌ATCC 43504几乎相同。限制性片段长度多态性(RFLP)分析表明,每只受感染的猴子携带1至4种菌株。在这22株分离物中发现的17种RFLP类型与其他16株分离物中发现的所有7种RFLP均不同。因此,与来自不同猴群的猴子相比,同一猴群中的猴子更有可能被具有相同或相似RFLP的幽门螺杆菌分离株感染。通过PCR扩增并从恒河猴85D08的幽门螺杆菌分离株中克隆了16S rRNA基因。用该探针进行核糖体分型显示,恒河猴分离株之间的多样性低于人类幽门螺杆菌分离株之间的多样性报道以及ureA-ureB基因簇PCR片段的RFLP分析结果。测定了克隆的16S rRNA基因的DNA序列,并与已报道的幽门螺杆菌和其他幽门螺杆菌物种的序列进行了比较。我们对127个核苷酸(对应于大肠杆菌16S rRNA基因的1240至1366位残基)的分析表明,来自猴子85D08的幽门螺杆菌分离株与人类幽门螺杆菌分离株的同源性为99.2%至100%,但在16S rRNA基因的该区域与其他幽门螺杆菌物种的同源性仅为83.5%至96.9%。这些数据为将这些分离株推定为幽门螺杆菌提供了有力支持。

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