Conville Patricia S, Brown June M, Steigerwalt Arnold G, Lee Judy W, Anderson Victoria L, Fishbain Joel T, Holland Steven M, Witebsky Frank G
Microbiology Service, Department of Laboratroy Medicine, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Bethesda, MD 20892-1508, USA.
J Clin Microbiol. 2004 Nov;42(11):5139-45. doi: 10.1128/JCM.42.11.5139-5145.2004.
Molecular methodologies have become useful techniques for the identification of pathogenic Nocardia species and for the recognition of novel species that are capable of causing human disease. Two isolates recovered from immunocompromised patients were characterized as Nocardia nova by biochemical and susceptibility testing results. The restriction fragment length polymorphism (RFLP) patterns obtained by restriction endonuclease analysis (REA) of an amplified portion of the heat shock protein gene were identical to those obtained with the type strain of N. nova. REA of an amplified portion of the 16S rRNA gene showed RFLP patterns that were unlike those obtained for the type strain of N. nova but that were similar to those obtained for the type strains of N. africana and N. veterana. Subsequent sequencing of a portion of the 16S rRNA gene produced identical results for the two patient isolates. Sequence analysis of 1,352-bp portions of the 16S rRNA gene indicated that these isolates were 99.8% similar to the recently described species N. veterana but were only 99.3, 98.1, and 98.1% similar to the type strains of N. africana, N. nova, and N. vaccinii, respectively. DNA-DNA hybridization studies confirmed that the two patient isolates belonged to the same species but were not closely related to N. africana, N. nova, N. vaccinii, or N. veterana. The patient isolates have been designated N. kruczakiae sp. nov. Because N. africana, N. veterana, and the new species are not readily differentiated from N. nova by phenotypic methods alone, the designation "N. nova complex" can be used to designate isolates such as these that phenotypically resemble N. nova but that have not been definitively characterized by 16S rRNA gene sequencing or DNA-DNA hybridization.
分子方法已成为鉴定致病性诺卡氏菌属菌种以及识别能够引起人类疾病的新菌种的有用技术。从免疫功能低下患者身上分离出的两株菌株,通过生化和药敏试验结果被鉴定为新星诺卡氏菌。通过对热休克蛋白基因扩增部分进行限制性内切酶分析(REA)获得的限制性片段长度多态性(RFLP)图谱,与新星诺卡氏菌模式菌株获得的图谱相同。对16S rRNA基因扩增部分进行的REA显示,其RFLP图谱与新星诺卡氏菌模式菌株的不同,但与非洲诺卡氏菌和兽医诺卡氏菌模式菌株的相似。随后对两株患者分离株的16S rRNA基因部分进行测序,结果相同。对16S rRNA基因1352 bp部分的序列分析表明,这些分离株与最近描述的兽医诺卡氏菌物种相似度为99.8%,但与非洲诺卡氏菌、新星诺卡氏菌和牛痘诺卡氏菌模式菌株的相似度分别仅为99.3%、98.1%和98.1%。DNA-DNA杂交研究证实,这两株患者分离株属于同一物种,但与非洲诺卡氏菌、新星诺卡氏菌、牛痘诺卡氏菌或兽医诺卡氏菌关系不密切。这两株患者分离株已被命名为克鲁扎克诺卡氏菌新种。由于仅通过表型方法难以将非洲诺卡氏菌、兽医诺卡氏菌和新物种与新星诺卡氏菌区分开来,因此“新星诺卡氏菌复合体”这一名称可用于指代那些表型上类似新星诺卡氏菌,但尚未通过16S rRNA基因测序或DNA-DNA杂交进行明确鉴定特征的分离株。