Wang He, Chen Ya-Lei, Teng Shih-Hua, Xu Zhi-Peng, Xu Ying-Chun, Hsueh Po-Ren
Department of Clinical Laboratory, Peking Union Medical College Hospital, Chinese Academy of Medical Sciences Beijing, China.
Department of Biotechnology, National Kaohsiung Normal University Kaohsiung, Taiwan.
Front Microbiol. 2016 Apr 8;7:415. doi: 10.3389/fmicb.2016.00415. eCollection 2016.
Burkholderia pseudomallei is not represented in the current version of Bruker Biotyper matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) system. A total of 66 isolates of B. pseudomallei, including 30 clinical isolates collected from National Taiwan University Hospital (NTUH, n = 27) and Peking Union Medical College Hospital (PUMCH, n = 3), and 36 isolates of genetically confirmed strains, including 13 from clinical samples and 23 from environmental samples, collected from southern Taiwan were included in this study. All these isolates were identified by partial 16S rDNA gene sequencing analysis and the Bruker Biotyper MALDI-TOF MS system. Among the 30 isolates initially identified as B. pseudomallei by conventional identification methods, one was identified as B. cepacia complex (NTUH) and three were identified as B. putida (PUMCH) by partial 16S rDNA gene sequencing analysis and Bruker Biotyper MALDI-TOF MS system. The Bruker Biotyper MALDI-TOF MS system misidentified 62 genetically confirmed B. pseudomallei isolates as B. thailandensis or Burkholderia species (score values, 1.803-2.063) when the currently available database (DB 5627) was used. However, using a newly created MALDI-TOF MS database (including B. pseudomallei NTUH-3 strain), all isolates were correctly identified as B. pseudomallei (score values >2.000, 100%). An additional 60 isolates of genetically confirmed B. cepacia complex and B. putida were also evaluated by the Bruker Biotyper MALDI-TOF MS system using the newly created database and none of these isolates were identified as B. pseudomallei. MALDI-TOF MS is a versatile and robust tool for the rapid identification of B. pseudomallei using the enhanced database.
伯克霍尔德菌在布鲁克生物分型基质辅助激光解吸/电离飞行时间质谱（MALDI-TOF MS）系统的当前版本中没有体现。本研究纳入了总共66株伯克霍尔德菌分离株，其中包括从台湾大学医院（NTUH，n = 27）和北京协和医院（PUMCH，n = 3）收集的30株临床分离株，以及从台湾南部收集的36株经基因确认的菌株，包括13株临床样本分离株和23株环境样本分离株。所有这些分离株均通过部分16S rDNA基因测序分析和布鲁克生物分型MALDI-TOF MS系统进行鉴定。在最初通过传统鉴定方法鉴定为伯克霍尔德菌的30株分离株中，通过部分16S rDNA基因测序分析和布鲁克生物分型MALDI-TOF MS系统，有1株被鉴定为洋葱伯克霍尔德菌复合体（NTUH），3株被鉴定为恶臭假单胞菌（PUMCH）。当使用当前可用数据库（DB 5627）时，布鲁克生物分型MALDI-TOF MS系统将62株经基因确认的伯克霍尔德菌分离株错误鉴定为泰国伯克霍尔德菌或伯克霍尔德菌属（得分值为1.803 - 2.063）。然而，使用新创建的MALDI-TOF MS数据库（包括伯克霍尔德菌NTUH-3菌株）时，所有分离株均被正确鉴定为伯克霍尔德菌（得分值>2.000，100%）。另外60株经基因确认的洋葱伯克霍尔德菌复合体和恶臭假单胞菌分离株也使用新创建的数据库通过布鲁克生物分型MALDI-TOF MS系统进行了评估，这些分离株均未被鉴定为伯克霍尔德菌。MALDI-TOF MS是一种使用增强数据库快速鉴定伯克霍尔德菌的通用且强大的工具。
