摘要
目的:建立志贺菌肠毒素1基因(ShET-1B)、志贺菌肠毒素2基因(ShET-2)、侵袭性质粒抗原H基因(ipaH)和侵袭相关基因(ial)的多重PCR鉴定方法,实现志贺菌多基因同时检测。方法:选择志贺菌的4种毒力基因作为靶基因,应用Touchdown PCR方法,在一个反应管中同时进行扩增,根据产物分子量定性判断结果。结果:多重PCR同时检测与单基因PCR分别检测4种毒力基因两种方法具有相同的灵敏度与特异性。624株志贺菌具有6种毒力基因型,571株福氏志贺菌(包括11个血清型)中556株ShET-1B阳性。结论:多重PCR鉴定志贺菌毒力基因具有简便、快速的特点,适用于毒力基因鉴定和流行病学调查研究。ShET-1B基因可能广泛存在于福氏志贺菌之中。
Objective：To develop a method of multiplex PCR to identify Shigella enterotoxins 1 gene （ShET- 1B）, Shigella enterotoxins 2 gene （ShET - 2）, the invasion plasmid antigen H gene （ipaH） and the invasion - associated locus （ial） in order to detect multi - gene of Shigella at the same time. Methods： Four kinds of virulence gene were selected as target genes and amplified at the same time in the reaction tube with Touchdown PCR. Then the result was decided based on molecular weight of the product. Results： Either 4 virulence genes were detected with multiplex PCR at the same time or detected separately with general PCR and it showed that both ways were sensitive and specific. Six genotypes of virulence gene were found in 624 strains of shigella and 556 strains were ShET - 1B positive of 571 strains of Shigella flexneri （ 11 serotypes included）. Conclusion：It is convenient and fast to determine virulence genes of shigella with multiplex PCR. The method is applicable to determine virulence genes for epidemiological investigations. ShET - 1B might abroadly exist in Shigella flexneri.
出处
《中国卫生检验杂志》
CAS
2006年第9期1038-1040,共3页
Chinese Journal of Health Laboratory Technology
