Polymerase chain reaction and an outer membrane protein gene probe for the detection of Porphyromonas gingivalis
Polymerase chain reaction and an outer membrane protein gene probe for the detection of Porphyromonas gingivalis
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DOI：
10.1111/j.1574-6968.1996.tb08151.x
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发表时间：
1996-05-01
影响因子：
2.1
通讯作者：
Abiko, Y
中科院分区：
文献类型：
--
作者：
Hiratsuka, K;Yoshida, W;Abiko, Y
建立了一种基于聚合酶链式反应(PCR)的牙龈卟啉单胞菌敏感性检测方法。扩增出426个碱基的DNA片段(278个碱基)，编码牙龈假单胞菌40 kDa的外膜蛋白。聚合酶链式反应产物来自所测试的牙龈假单胞菌菌株的染色体DNA，而不是来自其他口腔微生物的DNA。琼脂糖凝胶电泳法检测模板DNA的下限分别为10 pg/30个循环和100 fg/40个循环。将扩增产物与DRAI-HincII DNA片段进行杂交，检测下限分别为10 pg和10 fg的模板DNA，30次和40次。这些结果证明了该方法的简便、快速和特异性，以及Drai-HincII DNA片段在牙龈假单胞菌鉴定中的应用。
A sensitivity assay for Porphyromonas gingivalis based upon the polymerase chain reaction (PCR) was developed. A 426-bp sequence, including a DraI-HincII DNA fragment (278 bp) encoding the 40-kDa outer membrane protein of the P. gingivalis gene was amplified. PCR products were obtained from chromosomal DNAs of the P. gingivalis strains tested but nor from those of other oral microorganisms. The lower limit of template DNA detection was 10 pg with 30 cycles and 100 fg with 40 cycles of PCR by agarose gel electrophoresis. The PCR products were hybridized with DraI-HincII DNA fragment internal to the PCR primers regions used, The lower Limit of hybridization detection was 10 pg and 10 fg of template DNA with 30 and 40 cycles of PCR, respectively. These results demonstrated the simplicity, rapidity and specificity of the procedure, as well as the use of the DraI-HincII DNA fragment in the identification of P. gingivalis.