Ferroni A, Sermet-Gaudelus I, Abachin E, Quesnes G, Lenoir G, Berche P, Gaillard J L
Laboratoire de microbiologie, hôpital Necker-Enfants Malades, 149, rue de Sèvres, 75015, Paris, France.
Pathol Biol (Paris). 2003 Sep;51(7):405-11. doi: 10.1016/s0369-8114(03)00021-x.
We used partial 16S rRNA gene (16S DNA) sequencing for the prospective identification of nonfermenting Gram-negative bacilli recovered from patients attending our cystic fibrosis center (hôpital Necker-Enfants malades), which gave problematic results with conventional phenotypic tests. During 1999, we recovered 1093 isolates of nonfermenting Gram-negative bacilli from 702 sputum sampled from 148 patients. Forty-six of these isolates (27 patients) were not identified satisfactorily in routine laboratory tests. These isolates were identified by 16S DNA sequencing as Pseudomonas aeruginosa (19 isolates, 12 patients), Achromobacter xylosoxidans (10 isolates, 8 patients), Stenotrophomonas maltophilia (9 isolates, 9 patients), Burkholderia cepacia genomovar I/III (3 isolates, 3 patients), Burkholderia vietnamiensis (1 isolate), Burkholderia gladioli (1 isolate) and Ralstonia mannitolilytica (3 isolates, 2 patients). Fifteen isolates (33%) were resistant to all antibiotics in routine testing. Sixteen isolates (39%) resistant to colistin were recovered on B. cepacia-selective medium: 2 P. aeruginosa, 3 A. xylosoxidans, 3 S. maltophilia and the 8 Burkholderia--Ralstonia isolates. The API 20NE system gave no identification for 35 isolates and misidentified 11 isolates (2 P. aeruginosa, 2 A. xylosoxidans and 1 S. maltophilia classified as B. cepacia ). Control measures and/or treatment were clearly improved as a result of 16S DNA sequencing in three of these cases. This study confirms the weakness of phenotypic methods for identification of atypical nonfermenting Gram-negative bacilli recovered from cystic fibrosis patients. The genotypic methods, such as 16S DNA sequencing which allows identification of strains in routine practice, appears to have a small, but significant impact on the clinical management of CF patients.
我们采用部分16S rRNA基因（16S DNA）测序法对从我们囊性纤维化中心（内克尔儿童医院）就诊患者中分离出的非发酵革兰氏阴性杆菌进行前瞻性鉴定，这些细菌用传统表型试验检测结果存在问题。1999年期间，我们从148例患者的702份痰液样本中分离出1093株非发酵革兰氏阴性杆菌。其中46株（27例患者）在常规实验室检测中未能得到满意鉴定。通过16S DNA测序，这些菌株被鉴定为铜绿假单胞菌（19株，12例患者）、木糖氧化无色杆菌（10株，8例患者）、嗜麦芽窄食单胞菌（9株，9例患者）、洋葱伯克霍尔德菌基因Ⅰ/Ⅲ型（3株，3例患者）、越南伯克霍尔德菌（1株）、唐菖蒲伯克霍尔德菌（1株）和曼氏罗尔斯顿菌（3株，2例患者）。15株（33%）在常规检测中对所有抗生素耐药。在洋葱伯克霍尔德菌选择性培养基上分离出16株（39%）对黏菌素耐药的菌株：2株铜绿假单胞菌、3株木糖氧化无色杆菌、3株嗜麦芽窄食单胞菌以及8株伯克霍尔德菌-罗尔斯顿菌属菌株。API 20NE系统对35株菌株无法鉴定，对11株菌株鉴定错误（2株铜绿假单胞菌、2株木糖氧化无色杆菌和1株嗜麦芽窄食单胞菌被鉴定为洋葱伯克霍尔德菌）。在其中3例病例中，16S DNA测序显著改善了控制措施和/或治疗方案。本研究证实了表型方法在鉴定从囊性纤维化患者中分离出的非典型非发酵革兰氏阴性杆菌方面存在不足。基因型方法，如16S DNA测序，能够在常规实践中鉴定菌株，似乎对囊性纤维化患者的临床管理产生了虽小但显著的影响。
