Bavarian Health and Food Safety Authority, Oberschleißheim, Germany.
FEMS Microbiol Lett. 2012 Mar;328(1):46-53. doi: 10.1111/j.1574-6968.2011.02479.x. Epub 2012 Jan 11.
A real-time PCR procedure targeting the gene of the molecular cochaperon DnaJ (dnaJ) was developed for specific detection of strains belonging to the Enterobacter cloacae group. The inclusivity and exclusivity of the real-time PCR assay were assessed with seven reference strains of E. cloacae, 12 other Enterobacter species and 41 non-Enterobacter strains. Inclusivity as well as exclusivity of the duplex real-time PCR was 100%. In contrast, resolution of matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS) was inadequate for delineation of Enterobacter asburiae, Enterobacter hormaechei, Enterobacter kobei and Enterobacter ludwigii from E. cloacae. Eleven of 56 (20%) clinical isolates of the E. cloacae group could not be clearly identified as a certain species using MALDI-TOF MS. In summary, the combination of MALDI-TOF MS with the E. cloacae-specific duplex real-time PCR is an appropriate method for identification of the six species of the E. cloacae complex.
开发了一种针对分子伴侣 DnaJ（dnaJ）基因的实时 PCR 程序，用于特异性检测肠杆菌属 cloacae 组的菌株。用 7 株参考菌株、12 株其他肠杆菌属菌株和 41 株非肠杆菌属菌株评估了实时 PCR 检测的包容性和排他性。该实时 PCR 双管检测的包容性和排他性均为 100%。相比之下，基质辅助激光解吸电离飞行时间质谱（MALDI-TOF MS）的分辨率不足以区分 asburiae 肠杆菌、hormaechei 肠杆菌、kobei 肠杆菌和 ludwigii 肠杆菌与 cloacae 肠杆菌。使用 MALDI-TOF MS，56 株 cloacae 肠杆菌组临床分离株中 11 株（20%）无法明确鉴定为特定种。总之，MALDI-TOF MS 与 cloacae 特异性实时 PCR 双管检测的结合是鉴定 cloacae 肠杆菌复合体 6 个种的合适方法。
