Department of Clinical Laboratory, Peking Union Medical College Hospital, Chinese Academy of Medical Sciences, Beijing 100730, People’s Republic of China.
J Clin Microbiol. 2011 May;49(5):1890-8. doi: 10.1128/JCM.02415-10. Epub 2011 Mar 9.
Eleven reference and 25 clinical isolates of Fusarium were subject to multilocus DNA sequence analysis to determine the species and haplotypes of the fusarial isolates from Beijing and Shandong, China. Seven loci were analyzed: the translation elongation factor 1 alpha gene (EF-1α); the nuclear rRNA internal transcribed spacer (ITS), large subunit (LSU), and intergenic spacer (IGS) regions; the second largest subunit of the RNA polymerase gene (RPB2); the calmodulin gene (CAM); and the mitochondrial small subunit (mtSSU) rRNA gene. We also evaluated an IGS-targeted PCR/reverse line blot (RLB) assay for species/haplotype identification of Fusarium. Twenty Fusarium species and seven species complexes were identified. Of 25 clinical isolates (10 species), the Gibberella (Fusarium) fujikuroi species complex was the commonest (40%) and was followed by the Fusarium solani species complex (FSSC) (36%) and the F. incarnatum-F. equiseti species complex (12%). Six FSSC isolates were identified to the species level as FSSC-3+4, and three as FSSC-5. Twenty-nine IGS, 27 EF-1α, 26 RPB2, 24 CAM, 18 ITS, 19 LSU, and 18 mtSSU haplotypes were identified; 29 were unique, and haplotypes for 24 clinical strains were novel. By parsimony informative character analysis, the IGS locus was the most phylogenetically informative, and the rRNA gene regions were the least. Results by RLB were concordant with multilocus sequence analysis for all isolates. Amphotericin B was the most active drug against all species. Voriconazole MICs were high (>8 μg/ml) for 15 (42%) isolates, including FSSC. Analysis of larger numbers of isolates is required to determine the clinical utility of the seven-locus sequence analysis and RLB assay in species classification of fusaria.
从中国北京和山东采集的 11 株参考菌株和 25 株临床分离株进行多位点 DNA 序列分析，以确定这些真菌的种和单倍型。分析了 7 个基因座：翻译延伸因子 1α（EF-1α）基因；核 rRNA 内转录间隔区（ITS）、大亚基（LSU）和基因间间隔区（IGS）；RNA 聚合酶 II 大亚基（RPB2）基因；钙调蛋白基因（CAM）；线粒体小亚基（mtSSU）rRNA 基因。我们还评估了一种基于 IGS 的 PCR/反向线杂交（RLB）检测方法，用于鉴定 Fusarium 的种/单倍型。鉴定出 20 种 Fusarium 种和 7 个种复合体。在 25 株临床分离株（10 个种）中，最常见的是藤仓赤霉（Gibberella）种复合体（40%），其次是茄病镰刀菌（Fusarium）种复合体（36%）和 F. incarnatum-F. equiseti 种复合体（12%）。6 株 FSSC 分离株被鉴定为 FSSC-3+4，3 株被鉴定为 FSSC-5。鉴定出 29 个 IGS、27 个 EF-1α、26 个 RPB2、24 个 CAM、18 个 ITS、19 个 LSU 和 18 个 mtSSU 单倍型；29 个是独特的，24 个临床株的单倍型是新的。通过简约信息特征分析，IGS 基因座在系统发育上最具信息量，rRNA 基因区信息量最小。所有分离株的 RLB 结果与多基因座序列分析结果一致。两性霉素 B 是所有种中最有效的药物。15 株（42%）分离株的伏立康唑 MIC 值较高（>8μg/ml），包括 FSSC。需要对更多的分离株进行分析，以确定 7 个基因座序列分析和 RLB 检测在 Fusarium 种分类中的临床应用价值。
