Anderson Annette Carola, Sanunu Mohamed, Schneider Christian, Clad Andreas, Karygianni Lamprini, Hellwig Elmar, Al-Ahmad Ali
Department of Operative Dentistry and Periodontology, Albert-Ludwigs-University, Hugstetter Strasse 55, Freiburg, Germany.
Department of Hygiene and Microbiology, Albert-Ludwigs-University, Hermann-Herder-Str. 11, Freiburg, Germany.
BMC Microbiol. 2014 Dec 14;14:312. doi: 10.1186/s12866-014-0312-5.
Lactobacillus represents a large genus with different implications for the human host. Specific lactobacilli are considered to maintain vaginal health and to protect from urogenital infection. The presence of Lactobacillus species in carious lesions on the other hand is associated with progressive caries. Despite their clinical significance, species-level identification of lactobacilli still poses difficulties and mostly involves a combination of different phenotypic and genotypic methods. This study evaluated rapid MALDI-TOF MS analysis of vaginal and oral Lactobacillus isolates in comparison to 16S rDNA analysis.
Both methods were used to analyze 77 vaginal and 21 oral Lactobacillus isolates. The concordance of both methods was at 96% with five samples discordantly identified. Fifteen different Lactobacillus species were found in the vaginal samples, primarily L. iners, L. crispatus, L. jensenii and L. gasseri. In the oral samples 11 different species were identified, mostly L. salivarius, L. gasseri, L. rhamnosus and L. paracasei. Overall, the species found belonged to six different phylogenetic groups. For several samples, MALDI-TOF MS analysis only yielded scores indicating genus-level identification. However, in most cases the species found agreed with the 16S rDNA analysis result.
MALDI-TOF MS analysis proved to be a reliable and fast tool to identify lactobacilli to the species level. Even though some results were ambiguous while 16S rDNA sequencing yielded confident species identification, accuracy can be improved by extending reference databases. Thus, mass spectra analysis provides a suitable method to facilitate monitoring clinically relevant Lactobacillus species.
乳杆菌属是一个庞大的菌属,对人类宿主有不同影响。特定的乳杆菌被认为可维持阴道健康并预防泌尿生殖系统感染。另一方面,龋损部位存在乳杆菌与龋齿进展相关。尽管乳杆菌具有临床意义,但其种水平的鉴定仍存在困难,大多需要结合不同的表型和基因型方法。本研究评估了与16S rDNA分析相比,采用基质辅助激光解吸电离飞行时间质谱(MALDI-TOF MS)对阴道和口腔分离的乳杆菌进行快速分析。
两种方法用于分析77株阴道分离的乳杆菌和21株口腔分离的乳杆菌。两种方法的一致性为96%,有5个样本鉴定结果不一致。在阴道样本中发现了15种不同的乳杆菌,主要是惰性乳杆菌、卷曲乳杆菌、詹氏乳杆菌和加氏乳杆菌。在口腔样本中鉴定出11种不同的乳杆菌,主要是唾液乳杆菌、加氏乳杆菌、鼠李糖乳杆菌和副干酪乳杆菌。总体而言,所发现的菌种属于6个不同的系统发育组。对于几个样本,MALDI-TOF MS分析仅得出属水平鉴定的分数。然而,在大多数情况下,所发现的菌种与16S rDNA分析结果一致。
MALDI-TOF MS分析被证明是一种可靠且快速的将乳杆菌鉴定到种水平的工具。尽管在16S rDNA测序能明确鉴定菌种时,有些结果不明确,但通过扩展参考数据库可提高准确性。因此,质谱分析为促进监测临床相关乳杆菌种提供了一种合适的方法。