Conville Patricia S, Brown June M, Steigerwalt Arnold G, Lee Judy W, Byrer Dorothy E, Anderson Victoria L, Dorman Susan E, Holland Steven M, Cahill Barbara, Carroll Karen C, Witebsky Frank G
Microbiology Service, Department of Laboratory Medicine, Warren G. Magnuson Clinical Center, National Institutes of Health, U.S. Department of Health and Human Services, Bethesda, Maryland 20892, USA.
J Clin Microbiol. 2003 Jun;41(6):2560-8. doi: 10.1128/JCM.41.6.2560-2568.2003.
The molecular methodologies used in our laboratories have allowed us to define a group of Nocardia isolates from clinical samples which resemble the type strain of Nocardia veterana. Three patient isolates and the type strain of N. veterana gave identical and distinctive restriction fragment length polymorphisms (RFLPs) for an amplified portion of the 16S rRNA gene. These three isolates and the N. veterana type strain also gave identical RFLPs for an amplified portion of the 65-kDa heat shock protein gene, but this pattern was identical to that obtained for the Nocardia nova type strain. Sequence analysis of both a 1,359-bp region of the 16S rRNA gene and a 441-bp region of the heat shock protein gene of the patient isolates showed 100% identities with the same regions of the N. veterana type strain. DNA-DNA hybridization of the DNA of one of the patient isolates with the DNA of the N. veterana type strain showed a relative binding ratio of 82%, with 0% divergence, confirming that the isolate was N. veterana. Biochemical and susceptibility testing showed no significant differences among the patient isolates and the N. veterana type strain. Significantly, the results of antimicrobial susceptibility testing obtained for our isolates were similar to those obtained for N. nova, indicating that susceptibility testing alone cannot discriminate between these species. We present two case studies which show that N. veterana is a causative agent of pulmonary disease in immunocompromised patients residing in North America. We also describe difficulties encountered in using 16S rRNA gene sequences alone for discrimination of N. veterana from the related species Nocardia africana and N. nova because of the very high degree of 16S rRNA gene similarity among them.
我们实验室所使用的分子方法,使我们能够从临床样本中确定出一组诺卡氏菌分离株,这些分离株与诺卡氏菌旧种的模式菌株相似。三株患者分离株和诺卡氏菌旧种的模式菌株,在16S rRNA基因的一个扩增片段上呈现出相同且独特的限制性片段长度多态性(RFLP)。这三株分离株和诺卡氏菌旧种的模式菌株,在65-kDa热休克蛋白基因的一个扩增片段上也呈现出相同的RFLP,但这种模式与新星诺卡氏菌模式菌株所获得的模式相同。对患者分离株的16S rRNA基因的一个1359 bp区域和热休克蛋白基因的一个441 bp区域进行序列分析,结果显示与诺卡氏菌旧种模式菌株的相同区域具有100%的同一性。将其中一株患者分离株的DNA与诺卡氏菌旧种模式菌株的DNA进行DNA-DNA杂交,结果显示相对结合率为82%,差异为0%,证实该分离株为诺卡氏菌旧种。生化和药敏试验表明,患者分离株与诺卡氏菌旧种模式菌株之间没有显著差异。值得注意的是,我们分离株的抗菌药敏试验结果与新星诺卡氏菌的结果相似,这表明仅靠药敏试验无法区分这些菌种。我们展示了两个病例研究,表明诺卡氏菌旧种是北美免疫功能低下患者肺部疾病的病原体。我们还描述了仅使用16S rRNA基因序列来区分诺卡氏菌旧种与相关菌种非洲诺卡氏菌和新星诺卡氏菌时遇到的困难,因为它们之间的16S rRNA基因相似度非常高。