Sapico F L, Reeves D, Wexler H M, Duncan J, Wilson K H, Finegold S M
Medical and Research Services, Wadsworth Division, Veterans Administration Medical Center, West Los Angeles, California 90073.
J Clin Microbiol. 1994 Oct;32(10):2510-3. doi: 10.1128/jcm.32.10.2510-2513.1994.
Portions of the 16S RNA from a urease-positive Bilophila wadsworthia strain were sequenced, and a probe was constructed. The probe was end labeled with [32P]ATP and polynucleotide kinase and hybridized on a nylon filter (by dot blot hybridization) to the immobilized rRNA of 12 B. wadsworthia strains and eight other anaerobic isolates. The probe efficiently hybridized only to the Bilophila strains. Cross-reactivity at high RNA levels (2,000 ng) was observed with one strain of Bacteroides thetaiotamicron and one strain of Bacteroides fragilis (with 10x SET buffer [20x SET buffer is 0.5 M NaCl, 0.03 M Tris, and 2 mM EDTA]) but was not seen at lower RNA levels or with 5x SET buffer. When tested against mixed cultures of aerobic and anaerobic isolates representative of appendiceal abscess flora, the probe did not react with mixed cultures containing no Bilophila cells and could detect > or = 10(5) Bilophila CFU/ml when the mixture was seeded with Bilophila cells. This probe is of potential use in the rapid identification of pure isolates and in the direct identification of B. wadsworthia in clinical specimens.
对一株产尿素酶的沃兹沃思嗜胆菌（Bilophila wadsworthia）的16S RNA部分序列进行了测定，并构建了一个探针。该探针用[32P]ATP和多核苷酸激酶进行末端标记，并通过斑点印迹杂交在尼龙滤膜上与12株沃兹沃思嗜胆菌菌株及其他8株厌氧分离株固定化的rRNA进行杂交。该探针仅能有效地与嗜胆菌菌株杂交。在高RNA水平（2000 ng）下，观察到该探针与一株多形拟杆菌（Bacteroides thetaiotamicron）和一株脆弱拟杆菌（Bacteroides fragilis）有交叉反应（使用10倍SET缓冲液[20倍SET缓冲液为0.5 M NaCl、0.03 M Tris和2 mM EDTA]），但在较低RNA水平或使用5倍SET缓冲液时未观察到交叉反应。当针对代表阑尾脓肿菌群的需氧和厌氧分离株的混合培养物进行测试时，该探针与不含嗜胆菌细胞的混合培养物无反应，而当混合物接种嗜胆菌细胞时，该探针能够检测到≥10(5) CFU/ml的嗜胆菌。该探针在快速鉴定纯分离株以及直接鉴定临床标本中的沃兹沃思嗜胆菌方面具有潜在用途。
