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幽门螺杆菌基因异质性的脉冲场凝胶电泳分析及DNA同源性的重新评估

Genetic heterogeneity of Helicobacter pylori by pulse-field gel electrophoresis and re-evaluation of DNA homology.

作者信息

Takami S, Hayashi T, Akashi H, Shimoyama T, Tamura T

机构信息

Department of Bacteriology, Hyogo College of Medicine, Japan.

出版信息

Eur J Gastroenterol Hepatol. 1994 Dec;6 Suppl 1:S53-6.

PMID:7735937
Abstract

PURPOSE

The genetic heterogeneity of Helicobacter pylori isolates was re-evaluated by using pulse-field gel electrophoresis to examine macrorestriction patterns and by studying DNA homology.

MATERIALS AND METHODS

Twenty H. pylori isolates, two closely related species, H. mustelae and H. felis, and Campylobacter spp. were used. Notl-digested macrorestriction patterns were examined by pulse-field gel electrophoresis. DNA homology was examined by the S1 nuclease method, using [3H]-labeled DNA from H. pylori NCTC11637 and two H. pylori isolates for reference.

RESULT

Intergenus DNA homology between H. pylori and Campylobacters was 50-60%. Interspecies homology between H. pylori and H. mustelae or H. felis was around 60%. Intraspecies homology among H. pylori isolates was above 80%, except for a few that exhibited 70-80% homology. These findings indicate that all H. pylori isolates were homogeneous and belonged to the same species. Notl pulse-field gel electrophoresis patterns of H. pylori isolates differed markedly at the individual strain level. There was no specific relationship to any deviation from DNA homology, and the differences were observed within rather homogeneous members of the species.

CONCLUSION

The polymorphism in the Notl pulse-field gel electrophoresis patterns of H. pylori isolates differed markedly among strains, even though these isolates displayed species homogeneity, with DNA homology of 70-100%.

摘要

目的

通过使用脉冲场凝胶电泳检测宏观限制性酶切图谱并研究DNA同源性,对幽门螺杆菌分离株的遗传异质性进行重新评估。

材料与方法

使用了20株幽门螺杆菌分离株、两种密切相关的物种——鼬獾螺杆菌和猫螺杆菌以及弯曲杆菌属细菌。通过脉冲场凝胶电泳检测经NotI酶切后的宏观限制性酶切图谱。采用S1核酸酶法,以幽门螺杆菌NCTC11637和两株幽门螺杆菌分离株的[3H]标记DNA为参照检测DNA同源性。

结果

幽门螺杆菌与弯曲杆菌属之间的属间DNA同源性为50% - 60%。幽门螺杆菌与鼬獾螺杆菌或猫螺杆菌之间的种间同源性约为60%。幽门螺杆菌分离株之间的种内同源性高于80%,少数分离株的同源性为70% - 80%。这些结果表明,所有幽门螺杆菌分离株均具有同质性,属于同一物种。幽门螺杆菌分离株的NotI脉冲场凝胶电泳图谱在个体菌株水平上差异显著。与DNA同源性的任何偏差均无特定关系,且这些差异在该物种相当同质的成员中也可观察到。

结论

幽门螺杆菌分离株NotI脉冲场凝胶电泳图谱的多态性在菌株之间差异显著,尽管这些分离株显示出物种同质性,DNA同源性为70% - 100%。

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