Oligonucleotide probes to the 16S ribosomal RNA: implications of sequence homology and secondary structure with particular reference to the oral species Prevotella intermedia and Prevotella nigrescens.
Oligonucleotide probes to the 16S ribosomal RNA: implications of sequence homology and secondary structure with particular reference to the oral species Prevotella intermedia and Prevotella nigrescens.
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16S 核糖体 RNA 的寡核苷酸探针：序列同源性和二级结构的影响，特别是口腔物种中间普雷沃菌和黑质普雷沃菌。
DOI：
10.1111/j.1601-0825.1995.tb00154.x
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发表时间：
2008
期刊：
影响因子：
3.8
通讯作者：
S. Finegold
中科院分区：
文献类型：
--
作者：
H. Shah;S. Gharbia;C. Scully;S. Finegold
基于小亚基16S核糖体RNA基因序列区域的8个寡核苷酸在分子内的位置及其二维结构的背景下进行了分析，以使其在识别中间普雷沃氏菌和黑化普雷沃氏菌中的应用合理化。对41株口腔和呼吸道临床分离株及2株参考菌株进行DNA-DNA杂交和多位点酶电泳鉴定。将标记为I Bi-2至I Bi-6（用于P. intermedia）和2Bi-2（用于P. nigrescens）的寡核苷酸探针与16S rRNA比对表明，这些探针缺乏特异性或由高变区构建。一种52聚寡核苷酸（代号Bi）可靠地检测了这两种物种。由于两种物种的16S rnas高度一致，因此需要改变杂交条件的严格程度来检测两种物种。因此探针I Bi-I识别中间假体，而I Bi-I在低强度下同时检测中间假体和黑孢假体。然而，在高严格度条件下，探针2bi - 1只识别出P. nigresensis。这些探针具有高度特异性，不会与其他牙周病原体（如核梭杆菌、放线菌、齿状密螺旋体）和几种色素物种（如黑色素普雷沃氏菌、齿状假杆菌、loescheip、解糖卟啉单胞菌、Py）的DNA杂交。Py endodontalis。Py gingivalis。李维斯和Py。macacae。
Eight oligonucleotides based upon regions of the small subunit 16S ribosomal RNA gene sequences were analysed against a background of their position within the molecule and their two-dimensional structure to rationalise their use in recognising Prevotella intermedia and Prevotella nigrescens. The 41 clinical isolates from both oral and respiratory sites and two reference strains were subjected to DNA-DNA hybridisation and multilocus enzyme electrophoresis to confirm their identity. Alignment of oligonucleotide probes designated I Bi-2 to I Bi-6 (for P. intermedia) and 2Bi-2 (for P. nigrescens) with the 16S rRNA suggested that these probes lacked specificity or were constructed from hypervariable regions. A 52-mer oligonucleotide (designated Bi) reliably detected both species. Because of the high degree of concordance between the 16S rRNAs of both species, it was necessary to vary the stringency of hybridisation conditions for detection of both species. Thus probe I Bi-I recognised P. intermedia while I Bi-I detected both P. intermedia and P. nigrescens at low stringency. However, under conditions of high stringency only P. nigrescens was recognised by probe 2Bi-I. These probes were highly specific and did not hybridise with DNA from the closely related P. corporis, nor other periodontal pathogens such as Fusobacterium nucleatum, Actinobacillus actinomycetemcomitans, Treponema denticola and several pigmented species such as Prevotella melaninogenica, P. denticola, P. loescheii, Porphyromonas asaccharolytica, Py. endodontalis, Py. gingivalis, Py. levii, and Py. macacae.