Molecular epidemiological and antibiotic susceptibility characterization of Brucella isolates from humans in sicily, Italy
Molecular epidemiological and antibiotic susceptibility characterization of Brucella isolates from humans in sicily, Italy
复制标题
DOI：
10.1128/jcm.00822-07
复制
发表时间：
2007-09-01
影响因子：
9.4
通讯作者：
Ciuchini, Franco
中科院分区：
文献类型：
--
作者：
Marianelli, Cinzia;Graziani, Caterina;Ciuchini, Franco
布鲁氏菌病在西西里是一个严重的问题。在意大利，羊布鲁氏菌被确定为人类中最常见的分离种。然而，目前还没有关于人类布鲁氏菌分离株的分子流行病学特征的数据。本研究对临床分离的布鲁氏菌进行了分子特征研究。并评估它们的抗微生物敏感性。对20株布鲁氏菌进行了研究。利用不同的生长特性和DNA多态分析，如聚合酶链式反应扩增的omp2a和omp2b基因的限制性内切酶图谱、rpoB核苷酸序列分析和16个基因座的多位点可变数目串联重复序列分析(MLVA-16)来鉴定菌株的特征。用E-test法测定两种不同琼脂上的体外药敏结果，并对结果进行比较。RpoB核苷酸序列分析可以鉴定出两种不同的基因类型。另一方面，MLVA-16分型试验识别出17种不同的基因类型。所有菌株对所有受试抗生素(利福平、多西环素、环丙沙星、头孢曲松和新诺明-磺胺甲恶唑)均敏感，用E-test法推荐用Mueller-Hinton琼脂纸片进行药敏试验。我们的发现确定了西西里岛分离出的梅里特氏杆菌生物型3为病原菌，并鼓励在流行病学追溯分析中使用这两种分子方法，特别是MLVA-16方法。这项研究是在意大利，特别是在西西里岛东部流行的布鲁氏菌菌株分子分型的第一个流行病学数据。
Brucellosis is a serious problem in Sicily. Brucella melitensis was identified as the species most frequently isolated in humans in Italy. No data, however, are available about the molecular epidemiological characterization of Brucella isolates from humans. We have conducted this study to molecularly characterize clinical isolates of Brucella spp. and to evaluate their antimicrobial susceptibilities. Twenty Brucella isolates were studied. Differential growth characteristics and DNA polymorphisms such as the restriction patterns of the PCR-amplified omp2a and omp2b genes, rpoB nucleotide sequencing, and multiple-locus variable-number tandem repeat analysis of 16 loci (MLVA-16) were used to characterize the strains. In vitro antibiotic susceptibility was determined by the E-test method on two different agar media, and the results were compared. All isolates were identified as B. melitensis biovar 3. rpoB nucleotide sequence analysis allowed the identification of two different genotypes of B. melitensis biovar 3. On the other hand, the MLVA-16 typing assay recognized 17 distinct genotypes. All isolates were sensitive to all tested antibiotics (rifampin, doxycycline, ciprofloxacin, ceftriaxone, and trimethoprim-sulfamethoxazole), and the Mueller-Hinton agar plate is recommended for antibiotic susceptibility testing by the E-test method. Our findings identify B. melitensis biovar 3 as the etiological agent isolated in Sicily and encourage the use of both molecular methods, and in particular of the MLVA-16 assay, in epidemiological trace-back analysis. This study represents the first epidemiological data from molecular typing of Brucella strains circulating in Italy and, in particular, in eastern Sicily.