Use of checkerboard DNA-DNA hybridization to study complex microbial ecosystems
Use of checkerboard DNA-DNA hybridization to study complex microbial ecosystems
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DOI：
10.1111/j.1399-302x.2004.00168.x
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发表时间：
2004-12-01
影响因子：
--
通讯作者：
Goodson, JM
中科院分区：
文献类型：
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作者：
Socransky, SS;Haffajee, AD;Goodson, JM
使用传统的微生物学技术对微生物复杂的生态系统进行大规模的研究是困难的。新的探针-靶格式的分子鉴定技术，如棋盘式DNA-DNA杂交，允许在非常大量的样品中计数大量的物种。以棋盘杂交形式测试了40种常见牙龈下物种的洋地黄毒苷标记的全基因组探针。使用荧光成像仪定量杂交反应产生的化学荧光信号，并用于评价探针的灵敏度和特异性。调整DNA探针的灵敏度以检测10（4）个细胞。对80个可培养的物种，93.5%的潜在交叉反应显示出信号
It has been difficult to conduct large scale studies of microbiologically complex ecosystems using conventional microbiological techniques. Molecular identification techniques in new probe-target formats, such as checkerboard DNA-DNA hybridization, permit enumeration of large numbers of species in very large numbers of samples. Digoxigenin-labeled whole genomic probes to 40 common subgingival species were tested in a checkerboard hydridization format. Chemifluorescent signals resulting from the hybridization reactions were quantified using a Fluorimager and used to evaluate sensitivity and specificity of the probes. Sensitivity of the DNA probes was adjusted to detect 10(4) cells. In all, 93.5% of potential cross-reactions to 80 cultivable species exhibited signals