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目的探讨多重PCR技术用于检测牙龈卟啉单胞菌(Porphyromonasgingivalis,P.gingivalis)致病岛rag基因的可行性,并研究致病岛rag基因在毒力株和非毒力株中的分布情况。方法本研究于2011年1—6月在山东省口腔生物医学重点实验室进行,对P.gingivalis毒力株和非毒力株进行厌氧培养,采用普通PCR和多重PCR分别对他们的致病岛rag基因进行检测,对比检测结果,并对临床标本进行预实验研究。结果普通PCR和多重PCR均能对P.gingivalis致病岛rag基因进行检测,并且结果一致。多重PCR可用于临床标本的检测。致病岛rag基因不同基因型在毒力株和非毒力株中的分布不同,rag-1型存在于高毒力株P.gingivalisW83中,而rag-4型存在于低毒力株P.gingivalisATCC33277中。结论致病岛rag基因与细菌致病性密切相关;多重PCR技术省时省力、简单快速,为后续临床标本中rag基因的快速检测提供实验依据。

Detection of rag genotypes of Porphyromonas gingivalis with multiple PCR

Objective To optimize muhiple PCR reaction conditions and to detect rag genotypes of Porphyromonas gingivalis in the virulence strain and avirulent strains. Methods in Shandong Provincial Key Laboratory of Oral Biomedicine This study was carried out from January 2011 to June 2011 Anaerobic culture was done in the virulence strain and avirulent strains of Porphyromonas gingivalis, and detect the rag genotypes of Porphyromonas gingivalis with multiple PCR and common PCR. And optimize muhiple PCR reaction system for pre-experimental study of clinical specimens. Results Both common PCR and muhiple PCR could be used to detect pathogenic islands gene of Porphyromonas gingivalis, and results were consistent. Optimized multiple PCR could be used for the detection of elinical specimens. The distribution of pathogenic island rag gene of different genetic type in the virulence strain and avirulent strains was different. Rag-1 existed in highly toxic strainW83 and rag-4 in minimally toxie strain ATCC33277. Conclusion Pathogenic islands gene of Porphyromonas gingivalis is closely related to the bacterial virulence. Multiple PCR technology can save time and energy and is simple and quick, which provides experimental basis for subsequent quick detection of rag gene of clinical speciment.