Use of randomly cloned DNA fragments for the identification of oral spirochetes.
Use of randomly cloned DNA fragments for the identification of oral spirochetes.
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使用随机克隆的 DNA 片段来鉴定口腔螺旋体。
DOI：
10.1111/j.1399-302x.1991.tb00457.x
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发表时间：
1991
影响因子：
--
通讯作者：
Boehringer,H
中科院分区：
文献类型：
--
作者：
DiRienzo,JM;Cornell,S;Boehringer,H
制备用于检测和鉴定口腔螺旋体的4种可培养菌种的DNA探针，即齿垢密螺旋体、索氏密螺旋体、文森特密螺旋体和食果胶密螺旋体。为了获得探针序列，将从每个菌种内的代表性菌株中分离的染色体DNA克隆到大肠杆菌K-12中。筛选克隆的DNA片段与仅来自同源菌株的DNA杂交的能力。鉴定了几个这样的片段，并且当针对来自革兰氏阴性和革兰氏阳性口腔细菌的一系列DNA进行测试时显示出特异性。所选探针序列在T. denticola和T. socranskii，从而观察到限制性片段长度多态性（RFLP）。在T. socranskii，RFLP可用于区分3个已知亚种。2株T.在严格条件下，Vincentii未能与来自这些菌株中的每一个的基因组DNA交叉杂交。该杂交探针适用于临床分离株的鉴定。denticolaand可用于检测混合培养物中单个密螺旋体物种的存在。在此基础上，成功地将该探针用于T.牙垢蛋白未培养菌斑样品。
DNA probes were produced for the detection and identification of 4 cultivable species of oral spirochetes,Treponema denticola, Treponema socranskii, Treponema vincentii andTreponema pectinovorum.To obtain probe sequences, chromosomal DNA, isolated from representative strains within each species, was cloned in Escherichia coli K‐12. Cloned DNA fragments were screened for the ability to hybridize to DNA only from homologous strains. Several such fragments were identified and shown to be specific when tested against a series of DNAs from gram‐negative and gram‐positive oral bacteria. The selected probe sequences were semi‐conserved within strains ofT. denticola and T. socranskiisuch that restriction fragment length polymorphism (RFLP) was observed. In the case ofT. socranskii, RFLP was useful in distinguishing between the 3 known subspecies. Chromosomal DNA fragments from 2 strains ofT. vincentiifailed to cross‐hybridize, under stringent conditions, to genomic DNA from each of these strains. The hybridization probes were suitable for the identification of clinical isolates ofT. denticolaand could be used to detect the presence of individual Treponema species in mixed cultures. On this basis, the probes were used successfully to detectT. denticolain uncultured plaque samples.