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[地高辛配基与生物素标记用于DNA探针鉴定TEM型β-内酰胺酶编码基因的比较研究]

[Comparative study with digoxigenin and biotin markers for the DNA probe identification of genes coding for type TEM beta-lactamases].

作者信息

Hermida M, Gibert I, Barbé J, Roy C

机构信息

Facultad de Medicina (Departamento de Genética y Microbiología), Universidad Autónoma de Barcelona.

出版信息

Enferm Infecc Microbiol Clin. 1990 Oct;8(8):501-4.

PMID:2095262
Abstract

We have tested two non-isotopic labels: digoxigenin--11-dUTP and biotin-7-dATP for the detection of TEM-1 beta-lactamase gene with a TEM-1 probe by DNA:DNA hybridisation (using spot technique). The use of radioactive labels is inconvenient and not available to all the clinical bacteriological laboratories. The strains tested were: 16 on file (14 carriers of different types of beta-lactamase) and 19 clinical isolated strains. Previously, the plasmid beta-lactamase of these strains had been identified by analytical isoelectric focusing. Using the probe labeled with digoxigenin-11-dUTP, the correlation between results obtained by DNA:DNA hybridisation and those by analytical isoelectric focusing was excellent, with no false positives; with the biotin--7-dATP labeled probe, reading was difficult and some false positives were observed.

摘要

我们通过DNA:DNA杂交(采用斑点技术),用TEM-1探针检测TEM-1β-内酰胺酶基因时,测试了两种非同位素标记物:地高辛配基-11-dUTP和生物素-7-dATP。使用放射性标记物不方便,且并非所有临床细菌学实验室都能获得。所测试的菌株包括:存档的16株(14株携带不同类型β-内酰胺酶)和19株临床分离菌株。此前,这些菌株的质粒β-内酰胺酶已通过分析性等电聚焦进行鉴定。使用用地高辛配基-11-dUTP标记的探针时,DNA:DNA杂交结果与分析性等电聚焦结果之间的相关性极佳,无假阳性;使用生物素-7-dATP标记的探针时,读数困难,且观察到一些假阳性。

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