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使用寡核苷酸探针直接检测猕猴牙菌斑样本中的牙龈卟啉单胞菌。

Direct detection of Porphyromonas gingivalis in Macaca fascicularis dental plaque samples using an oligonucleotide probe.

作者信息

Moncla B J, Braham P H, Persson G R, Page R C, Weinberg A

机构信息

Dental Diagnostics, Inc., Seattle, WA.

出版信息

J Periodontol. 1994 May;65(5):398-403. doi: 10.1902/jop.1994.65.5.398.

Abstract

Oligonucleotide probes complementary to the hypervariable regions of the 16S rRNA of Porphyromonas gingivalis, and previously shown to specifically identify human P. gingivalis strains to the species level, were tested for their ability to recognize P. gingivalis from nonhuman primates (Macaca fascicularis), either as distinct isolates or in subgingival dental plaque. The 32P-labeled probes hybridized with all 147 monkey isolates identified as P. gingivalis by morphology and biochemistry, but did not hybridize with any of the 331 isolates representing 17 genera of bacteria unrelated to P. gingivalis, or to the more closely related P. endodontalis and P. asaccharolytica. This corresponds to sensitivities and specificities of 100%. Of 76 M. fascicularis plaque samples, P. gingivalis was detected by probe and culture in 67. Of 26 human plaque samples taken from separate individuals free of periodontal disease, 23 failed to demonstrate P. gingivalis by probe or culture. The results of the combined 102 monkey and human plaque samples indicate that, when compared to culture as the "gold standard," the P. gingivalis probe had a sensitivity of 96%, a specificity of 87%, and an overall agreement with culture of 93%. These results reveal that the oligonucleotide probes used to identify P. gingivalis are specific for this organism, and give results comparable to culture methods for detecting the presence of P. gingivalis in M. fascicularis dental plaque.

摘要

与牙龈卟啉单胞菌16S rRNA高变区互补的寡核苷酸探针,先前已证明能在种水平特异性鉴定人类牙龈卟啉单胞菌菌株,现测试其识别非人灵长类动物(食蟹猴)牙龈卟啉单胞菌的能力,这些菌株可以是单独分离株,也可以是龈下牙菌斑中的菌株。32P标记的探针与通过形态学和生物化学鉴定为牙龈卟啉单胞菌的所有147株猴分离株杂交,但不与代表17个与牙龈卟啉单胞菌无关的细菌属的331株分离株杂交,也不与亲缘关系更近的牙髓卟啉单胞菌和解糖卟啉单胞菌杂交。这对应于100%的敏感性和特异性。在76份食蟹猴菌斑样本中,通过探针和培养检测到67份有牙龈卟啉单胞菌。在从无牙周病的不同个体采集的26份人类菌斑样本中,23份通过探针或培养未显示有牙龈卟啉单胞菌。102份猴和人类菌斑样本的综合结果表明,与作为“金标准”的培养法相比,牙龈卟啉单胞菌探针的敏感性为96%,特异性为87%,与培养法的总体一致性为93%。这些结果表明,用于鉴定牙龈卟啉单胞菌的寡核苷酸探针对该菌具有特异性,并且在检测食蟹猴牙菌斑中牙龈卟啉单胞菌的存在方面,其结果与培养方法相当。

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