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对来自地理和流行病学上不同来源的烟曲霉分离株的全细胞DNA进行限制性内切酶分析。

Restriction endonuclease analysis of total cellular DNA of Aspergillus fumigatus isolates of geographically and epidemiologically diverse origin.

作者信息

Denning D W, Clemons K V, Hanson L H, Stevens D A

机构信息

Department of Medicine, Santa Clara Valley Medical Center, San Jose, CA 95128.

出版信息

J Infect Dis. 1990 Nov;162(5):1151-8. doi: 10.1093/infdis/162.5.1151.

Abstract

No typing system exists for Aspergillus fumigatus, though isolates are distinguishable by phenotypic characteristics. DNA was prepared by lysis of protoplasts, followed by deproteination, phenolchloroform extraction, and dialysis. DNA prepared was of uniform size and exceeded 60 kb. After digestion with SalI and XhoI endonucleases, DNA was electrophoresed, stained, and photographed. Differences in the mobilities of 10- to 50-kb bands distinguished isolates. Reproducibility was shown by repeated preparations and animal passage. By use of a proposed notation system for describing restriction fragment length polymorphism patterns, 31 epidemiologically characterized isolates from three continents revealed 24 patterns (DNA types). Three DNA types were represented by 3 isolates each and 1 DNA type by 2 isolates; 20 types were unique. Two groups of 3 isolates of the same DNA type were from Stanford University Hospital. One patient isolate from Stanford was the same DNA type as a sewage isolate from New Jersey. Another Stanford isolate was the same as a German isolate. These observations indicate widespread dispersal of some clones and restricted locales for others. Paired isolates from airway fluids of three patients had two DNA types in each. Restriction endonuclease typing shows promise for investigating the epidemiology and ecology of A. fumigatus.

摘要

虽然烟曲霉不存在分型系统,但可通过表型特征区分分离株。通过原生质体裂解制备DNA,随后进行脱蛋白、酚氯仿抽提和透析。制备的DNA大小均一,超过60 kb。用SalI和XhoI内切酶消化后,对DNA进行电泳、染色和拍照。10至50 kb条带迁移率的差异可区分分离株。通过重复制备和动物传代显示了可重复性。使用一种提议的用于描述限制性片段长度多态性模式的符号系统,来自三大洲的31株经流行病学特征分析的分离株显示出24种模式(DNA型)。三种DNA型各由3株分离株代表,一种DNA型由2株分离株代表;20种类型是独特的。两组相同DNA型的3株分离株来自斯坦福大学医院。斯坦福大学的一株患者分离株与新泽西州的一株污水分离株DNA型相同。另一株斯坦福分离株与一株德国分离株相同。这些观察结果表明,一些克隆广泛传播,而另一些则局限于特定区域。来自三名患者气道分泌物的配对分离株各有两种DNA型。限制性内切酶分型在研究烟曲霉的流行病学和生态学方面显示出前景。

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