Qiu Y S, Klitorinos A, Rahal M D, Siboo R, Chan E C
Department of Microbiology and Immunology, Faculty of Medicine, McGill University, Montreal, Quebec, Canada.
Oral Microbiol Immunol. 1994 Oct;9(5):301-4. doi: 10.1111/j.1399-302x.1994.tb00075.x.
We recently developed a successful method for quantifying oral anaerobic spirochetes in pure culture by a viable count. New oral spirochete medium was used with low temperature-gelling agarose in polystyrene tissue-culture flasks. We have extended the use of this method to determine the viable count of spirochetes from periodontal pockets. Sixteen subgingival plaque samples were obtained by insertion of sterile paper points into deep periodontal pockets. The points were placed into reduced transport medium at chairside, vortexed in the microbiology laboratory and aliquots of the medium inoculated into molten new oral spirochete-agarose medium (37 degrees C) containing rifampin (20 micrograms/ml) in a flask. Subsequent dilutions were made from this initial flask to other flasks containing selective medium in sequence. All flasks were incubated anaerobically. Most other subgingival bacteria were selectively inhibited by rifampin. Spirochete colonies were typically spherical and were either dense or cottony. Their identities were checked by darkfield examination. Counts of colony-forming units of cultivable spirochetes ranged from 12.5% to 28.2% of the total cultivable anaerobic flora by the method described.
我们最近开发了一种通过活菌计数对纯培养中的口腔厌氧螺旋体进行定量的成功方法。在聚苯乙烯组织培养瓶中使用新型口腔螺旋体培养基与低温凝胶化琼脂糖。我们已将此方法的应用扩展到测定牙周袋中螺旋体的活菌计数。通过将无菌纸尖插入深部牙周袋来获取16份龈下菌斑样本。这些纸尖在椅旁放入还原运输培养基中，在微生物实验室中涡旋，然后将培养基的等分试样接种到含有利福平（20微克/毫升）的熔化的新型口腔螺旋体 - 琼脂糖培养基（37℃）的烧瓶中。随后从这个初始烧瓶依次向其他含有选择性培养基的烧瓶进行稀释。所有烧瓶均进行厌氧培养。大多数其他龈下细菌被利福平选择性抑制。螺旋体菌落通常呈球形，要么致密要么呈棉絮状。通过暗视野检查来确认它们的身份。按照所述方法，可培养螺旋体的菌落形成单位计数占可培养厌氧菌群总数的12.5％至28.2％。
