Cangelosi G A, Iversen J M, Zuo Y, Oswald T K, Lamont R J
MicroProbe Corporation, Diagnostics Division, Bothell, WA 98021.
Mol Cell Probes. 1994 Feb;8(1):73-80. doi: 10.1006/mcpr.1994.1011.
Oligodeoxyribonucleotide probes complementary to hypervariable regions of the 16S ribosomal RNA were designed for specificity toward Streptococcus mutans and Streptococcus sobrinus. The probes were tested for specificity and sensitivity by hybridization with nucleic acid from over 100 mutans and non-mutans oral streptococci and other common oropharyngeal bacteria. Probes designated SM002 and SM010 were 100% sensitive and > 99% specific for S. mutans. Probe SSP001, designed to detect both S. mutans and S. sobrinus, was 88% sensitive and > 99% specific. The probes were able to detect nucleic acid extracted from 3 x 10(4)-1 x 10(5) homologous bacteria. Sensitivity did not vary significantly with the growth state of the cells, except for diminished signals when using nucleic acid extracted from very old cultures. The probes correctly identified 72 S. mutans colonies isolated from 10 volunteer saliva samples, using sugar utilization patterns as a reference standard. Ten isolates resembling S. mutans by colony morphology but not by sugar utilization patterns were correctly distinguished from mutans streptococci by the probes. These results demonstrate that oligonucleotide probes can accurately identify S. mutans in saliva samples.
设计了与16S核糖体RNA高变区互补的寡脱氧核糖核苷酸探针，以特异性识别变形链球菌和远缘链球菌。通过与100多种变形链球菌和非变形链球菌口腔链球菌以及其他常见口咽细菌的核酸杂交，对这些探针的特异性和敏感性进行了测试。命名为SM002和SM010的探针对变形链球菌的敏感性为100%，特异性>99%。设计用于同时检测变形链球菌和远缘链球菌的探针SSP001，敏感性为88%，特异性>99%。这些探针能够检测从3×10⁴-1×10⁵同源细菌中提取的核酸。除了使用从非常陈旧培养物中提取的核酸时信号减弱外，敏感性随细胞生长状态的变化不显著。以糖利用模式作为参考标准，这些探针正确鉴定了从10名志愿者唾液样本中分离出的72个变形链球菌菌落。通过菌落形态类似于变形链球菌但糖利用模式不同的10个分离株，被这些探针与变形链球菌正确区分开来。这些结果表明，寡核苷酸探针可以准确识别唾液样本中的变形链球菌。
