基于基因探针，利用聚合酶链反应检测E型肉毒杆菌神经毒素结合蛋白

Gene probe-based detection of type E botulinum neurotoxin binding protein using polymerase chain reaction.

作者信息

Singh B R, Barcomb-Caddle L A, Fu F N, Li B

机构信息

Department of Chemistry, University of Massachusetts, Dartmouth 02747, USA.

出版信息

Toxicon. 1996 Jul;34(7):737-42. doi: 10.1016/0041-0101(95)00166-2.

DOI:10.1016/0041-0101(95)00166-2

PMID:8843574

Abstract

Using primers based on the nucleotide sequence of a neurotoxin binding protein from Type E Clostridium botulinum cultures, an amplified DNA product was obtained through polymerase chain reaction. The 400 base pair amplified DNA fragment was detectable with as low as 0.1 pg template DNA from Type E C. botulinum, and its fidelity was confirmed by Southern blotting using a DNA probe designed to detect the expected amplified DNA fragment. On the other hand, no DNA amplification was observed with as high as 10 ng template DNA from related Types A and B C. botulinum or from C. tetani, indicating the specificity of the probe.

摘要

利用基于来自E型肉毒梭菌培养物的神经毒素结合蛋白核苷酸序列设计的引物，通过聚合酶链反应获得了扩增的DNA产物。该400个碱基对的扩增DNA片段在低至0.1 pg的E型肉毒梭菌模板DNA中即可检测到，并且通过使用设计用于检测预期扩增DNA片段的DNA探针进行Southern印迹分析，证实了其保真度。另一方面，来自相关的A型和B型肉毒梭菌或破伤风梭菌的高达10 ng的模板DNA均未观察到DNA扩增，表明该探针具有特异性。