Kim Yeon-Hee, Lee Si Young
Department of Microbiology and Immunology, College of Dentistry, Research Institute of Oral Science, Gangneung-Wonju National University, Gangneung 210-702, Republic of Korea.
Department of Microbiology and Immunology, College of Dentistry, Research Institute of Oral Science, Gangneung-Wonju National University, Gangneung 210-702, Republic of Korea.
Arch Oral Biol. 2015 Feb;60(2):267-71. doi: 10.1016/j.archoralbio.2014.11.002. Epub 2014 Nov 8.
Mitis-salivarius (MS) agar has been used widely in microbial epidemiological studies because oral viridans streptococci can be selectively grown on this medium. Even though the previous findings reported the limited selecting power of MS agar for streptococcus strains, the identities of non-streptococcal strains from human oral samples which can grow on this medium are not clear yet. In this study, we identified non-streptococcal organisms grown on MS agar plates by polymerase chain reaction (PCR) amplification and sequencing of the 16S ribosomal RNA (rRNA) gene.
Eighty bacterial colonies on MS plates were isolated from plaque samples, and bacterial identification was achieved with the rapid ID 32 Strep system and mini API reader. The bacterial colonies identified as non-streptococci by the API system were selected for further identification. The 16S rRNA gene was amplified by PCR and verified using DNA sequencing analysis for identification. Sequences were compared with those of reference organisms in the genome database of the National Center for Biotechnology Information using the Basic Local Alignment Search Tool (BLAST).
Among the 11 isolated non-streptococcal strains on MS plates, 3 strains were identified as Actinomyces naeslundii, 7 strains were identified as Actinomyces oris and 1 strain were identified as Actinomyces sp. using Blastn.
In this study, we showed that some oral Actinomyces species can grow on Streptococcus-selective MS agar plates.
轻唾琼脂已广泛应用于微生物流行病学研究,因为口腔甲型溶血性链球菌能在此培养基上选择性生长。尽管先前的研究结果报道了轻唾琼脂对链球菌菌株的选择能力有限,但在这种培养基上生长的来自人类口腔样本的非链球菌菌株的种类尚不清楚。在本研究中,我们通过聚合酶链反应(PCR)扩增和16S核糖体RNA(rRNA)基因测序,鉴定了在轻唾琼脂平板上生长的非链球菌微生物。
从牙菌斑样本中分离出80个轻唾琼脂平板上的细菌菌落,并用快速ID 32 Strep系统和微型API读数器进行细菌鉴定。选择API系统鉴定为非链球菌的细菌菌落进行进一步鉴定。通过PCR扩增16S rRNA基因,并使用DNA测序分析进行验证以进行鉴定。使用基本局部比对搜索工具(BLAST)将序列与美国国立生物技术信息中心基因组数据库中的参考生物序列进行比较。
在轻唾琼脂平板上分离出的11株非链球菌菌株中,使用Blastn鉴定出3株为内氏放线菌,7株为口腔放线菌,1株为放线菌属。
在本研究中,我们表明一些口腔放线菌物种可以在选择性链球菌的轻唾琼脂平板上生长。