Kawamura Y, Nagase K, Endo M, Shigematsu T, Ezaki T, Yabuuchi E
Department of Microbiology, Gifu University School of Medicine, Japan.
Nihon Saikingaku Zasshi. 1991 May;46(3):597-610. doi: 10.3412/jsb.46.597.
A new simplified test kit, STAPHYOGRAM plate, was developed for 4-hr identification of aerobic, Gram-positive and catalase-positive cocci. The plate has 18 wells, in which different dehydrated substrates and nutrients are fixed. An 18-hr agar-culture suspension of a test strain with a turbidity of McFarland No. 4 was distributed into all wells in 50-microliters quantities. After 4-hr incubation at 37C, the profile number was obtained by summarizing positive reactions. The ability of the plate to differentiate the type strains of the 30 species of the three genera in the family Micrococcaceae was confirmed. These three genera are Staphylococcus, Micrococcus and Stomatococcus. The applicability of the fluorometric microplate hybridization technique to identification of aerobic, Gram-positive and catalase-positive cocci was confirmed by homologous hybridization among the type strains of the 30 species. Thus, 386 isolates of human and animal origin were pre-identified by microplate hybridization and used for evaluating the STAPHYOGRAM plate. Of the 236 profile numbers thus obtained with the 386 isolates, 218 (92.4%) were species-proper each and all for the 15 species of Staphylococcus and Stomatococcus mucilaginosus. A total of 342 (88.6%) of the 386 isolates were given such profile numbers, and were identified without any additional test. Among the 15 species identified primarily by the results of STAPHYOGRAM plate culture, S. caprae, S. lugdunensis, S. gallinarum and S. delphini were validly published after Approved Lists of Bacterial Names. The identified strains of S. caprae (48), S. haemolyticus (46), S. capitis (35) numbered between those of S. epidermidis (67) and S. saprophyticus (31). Profile numbers common to two species were seven (27 strains) and that to four species was one (17 strains). These 44 strains were identified with one to three additional tests. From these results, we were convinced that the STAPHYOGRAM test plate is useful for the rapid identification of members of family Micrococcaceae. By compiling STAPHYOGRAM plate data on genetically identified strains, an exclusive list of profile numbers will soon be prepared for perfection of the kit.
开发了一种新型简化检测试剂盒——葡萄球菌谱板,用于需氧、革兰氏阳性和过氧化氢酶阳性球菌的4小时鉴定。该板有18个孔,其中固定了不同的脱水底物和营养物质。将浊度为麦氏比浊法4号的测试菌株18小时琼脂培养物悬液以50微升的量分配到所有孔中。在37℃孵育4小时后,通过汇总阳性反应获得谱号。证实了该板区分微球菌科三个属30个种的模式菌株的能力。这三个属是葡萄球菌属、微球菌属和口腔球菌属。通过30个种的模式菌株之间的同源杂交,证实了荧光微孔板杂交技术在需氧、革兰氏阳性和过氧化氢酶阳性球菌鉴定中的适用性。因此,对386株人和动物来源的分离株进行了微孔板杂交预鉴定,并用于评估葡萄球菌谱板。用这386株分离株得到的236个谱号中,葡萄球菌属15个种和黏性口腔球菌的218个(92.4%)谱号与各自种完全匹配。386株分离株中共有342个(88.6%)获得了这样的谱号,无需任何额外测试即可鉴定。在主要根据葡萄球菌谱板培养结果鉴定的15个种中,山羊葡萄球菌、路邓葡萄球菌、鸡葡萄球菌和海豚葡萄球菌在《细菌名称认可名录》之后有效发表。鉴定出的山羊葡萄球菌(48株)、溶血葡萄球菌(46株)、头葡萄球菌(35株)的数量介于表皮葡萄球菌(67株)和腐生葡萄球菌(31株)之间。两个种共有的谱号有7个(27株),四个种共有的谱号有1个(17株)。这44株通过一到三项额外测试进行鉴定。从这些结果来看,我们确信葡萄球菌谱板检测对于微球菌科成员的快速鉴定是有用的。通过汇编基因鉴定菌株的葡萄球菌谱板数据,不久将编制一份专属谱号清单,以使试剂盒更加完善。