[艰难梭菌毒素A和毒素B基因3'末端重复片段的聚合酶链反应克隆]
[PCR cloning of 3'-terminal repeat fragments of Clostridium difficile toxin A and toxin B gene].
作者信息
Yang Xiao-qiang, Jiang Bo, Sun Yong, Wang Ji-de
机构信息
Institute of Digestive Diseases of PLA, Nanfang Hospital, First Military Medical University, Guangzhou 510515, China.
出版信息
Di Yi Jun Yi Da Xue Xue Bao. 2002 Sep;22(9):791-3.
OBJECTIVE
To explore a method to rapidly detect the presence of Clostridium difficile.
METHODS
PCR was used to amplify DNA fragments from the highly conserved and repeated domains of the 3'-terminal of Clostridium difficile toxin A gene and toxin B genes.
RESULTS
The fragments 960 bp and 1,851 bp in length were respectively amplified from toxin A and toxin B, while the control bacteria presented no distinct results.
CONCLUSION
The 960 bp and 1,851 bp fragments are specific for Clostridium difficile and PCR can be used to detect Clostridium difficile from the stool. As the peptides encoded by these conserved domains have high hydrophobicity and strong antigenicity, manufacture of the protein vaccine by gene engineering is possible on the basis of these conserved domains.
目的
探索一种快速检测艰难梭菌存在的方法。
方法
采用聚合酶链反应（PCR）从艰难梭菌毒素A基因和毒素B基因3′末端的高度保守且重复的结构域扩增DNA片段。
结果
分别从毒素A和毒素B中扩增出长度为960 bp和1851 bp的片段，而对照细菌未呈现明显结果。
结论
960 bp和1851 bp的片段对艰难梭菌具有特异性，PCR可用于从粪便中检测艰难梭菌。由于这些保守结构域编码的肽具有高疏水性和强抗原性，基于这些保守结构域通过基因工程制备蛋白疫苗是可行的。
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