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Molecular Epidemiological and Antibiotic Susceptibility Characterization of Brucella Isolates from Humans in Sicily, Italy
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 molecularl...
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Published in: | Journal of Clinical Microbiology 2007-09, Vol.45 (9), p.2923-2928 |
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creator | Marianelli, Cinzia Graziani, Caterina Santangelo, Carmela Xibilia, Maria Teresa Imbriani, Alida Amato, Rosa Neri, Domenico Cuccia, Mario Rinnone, Sebastiano Di Marco, Vincenzo Ciuchini, Franco |
description | 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. |
doi_str_mv | 10.1128/JCM.00822-07 |
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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.</description><identifier>ISSN: 0095-1137</identifier><identifier>EISSN: 1098-660X</identifier><identifier>EISSN: 1098-5530</identifier><identifier>DOI: 10.1128/JCM.00822-07</identifier><identifier>PMID: 17634297</identifier><identifier>CODEN: JCMIDW</identifier><language>eng</language><publisher>Washington, DC: American Society for Microbiology</publisher><subject>Anti-Bacterial Agents - pharmacology ; Bacterial Proteins - genetics ; Bacterial Typing Techniques ; Bacteriology ; Biological and medical sciences ; Brucella melitensis ; Brucella melitensis - classification ; Brucella melitensis - drug effects ; Brucella melitensis - genetics ; Brucella melitensis - isolation & purification ; Brucellosis - epidemiology ; Brucellosis - microbiology ; DNA, Bacterial - genetics ; DNA-Directed RNA Polymerases - genetics ; Epidemiology ; Fundamental and applied biological sciences. Psychology ; Genotype ; Humans ; Microbial Sensitivity Tests - methods ; Microbiology ; Miscellaneous ; Molecular Epidemiology ; Polymorphism, Genetic ; Polymorphism, Restriction Fragment Length ; Porins - genetics ; Sequence Analysis, DNA ; Sicily - epidemiology ; Tandem Repeat Sequences - genetics</subject><ispartof>Journal of Clinical Microbiology, 2007-09, Vol.45 (9), p.2923-2928</ispartof><rights>2007 INIST-CNRS</rights><rights>Copyright © 2007, American Society for Microbiology 2007</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c493t-d9380bbce79cd201ab44db06cf94eb6f45a45f08e200ff8be7aa9a674d8f85863</citedby><cites>FETCH-LOGICAL-c493t-d9380bbce79cd201ab44db06cf94eb6f45a45f08e200ff8be7aa9a674d8f85863</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://www.ncbi.nlm.nih.gov/pmc/articles/PMC2045243/pdf/$$EPDF$$P50$$Gpubmedcentral$$H</linktopdf><linktohtml>$$Uhttps://www.ncbi.nlm.nih.gov/pmc/articles/PMC2045243/$$EHTML$$P50$$Gpubmedcentral$$H</linktohtml><link.rule.ids>230,314,727,780,784,885,3188,3189,27924,27925,53791,53793</link.rule.ids><backlink>$$Uhttp://pascal-francis.inist.fr/vibad/index.php?action=getRecordDetail&idt=19093711$$DView record in Pascal Francis$$Hfree_for_read</backlink><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/17634297$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Marianelli, Cinzia</creatorcontrib><creatorcontrib>Graziani, Caterina</creatorcontrib><creatorcontrib>Santangelo, Carmela</creatorcontrib><creatorcontrib>Xibilia, Maria Teresa</creatorcontrib><creatorcontrib>Imbriani, Alida</creatorcontrib><creatorcontrib>Amato, Rosa</creatorcontrib><creatorcontrib>Neri, Domenico</creatorcontrib><creatorcontrib>Cuccia, Mario</creatorcontrib><creatorcontrib>Rinnone, Sebastiano</creatorcontrib><creatorcontrib>Di Marco, Vincenzo</creatorcontrib><creatorcontrib>Ciuchini, Franco</creatorcontrib><title>Molecular Epidemiological and Antibiotic Susceptibility Characterization of Brucella Isolates from Humans in Sicily, Italy</title><title>Journal of Clinical Microbiology</title><addtitle>J Clin Microbiol</addtitle><description>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.</description><subject>Anti-Bacterial Agents - pharmacology</subject><subject>Bacterial Proteins - genetics</subject><subject>Bacterial Typing Techniques</subject><subject>Bacteriology</subject><subject>Biological and medical sciences</subject><subject>Brucella melitensis</subject><subject>Brucella melitensis - classification</subject><subject>Brucella melitensis - drug effects</subject><subject>Brucella melitensis - genetics</subject><subject>Brucella melitensis - isolation & purification</subject><subject>Brucellosis - epidemiology</subject><subject>Brucellosis - microbiology</subject><subject>DNA, Bacterial - genetics</subject><subject>DNA-Directed RNA Polymerases - genetics</subject><subject>Epidemiology</subject><subject>Fundamental and applied biological sciences. Psychology</subject><subject>Genotype</subject><subject>Humans</subject><subject>Microbial Sensitivity Tests - methods</subject><subject>Microbiology</subject><subject>Miscellaneous</subject><subject>Molecular Epidemiology</subject><subject>Polymorphism, Genetic</subject><subject>Polymorphism, Restriction Fragment Length</subject><subject>Porins - genetics</subject><subject>Sequence Analysis, DNA</subject><subject>Sicily - epidemiology</subject><subject>Tandem Repeat Sequences - genetics</subject><issn>0095-1137</issn><issn>1098-660X</issn><issn>1098-5530</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2007</creationdate><recordtype>article</recordtype><recordid>eNqFkc1vEzEQxVcIRNPCjTOYA5y6Zbz2fviCVKJCg1pxCJW4WbNeO3HlXQd7lyr963GaiMKJkzXyT2_em5dlryicUVo0H77Or88AmqLIoX6SzSiIJq8q-PE0mwGIMqeU1UfZcYy3AJTzsnyeHdG6YrwQ9Sy7v_ZOq8lhIBcb2-neeudXVqEjOHTkfBhta_1oFVlOUenNbnR23JL5GgOqUQd7j6P1A_GGfAqT0s4hWUTvcNSRmOB7cjn1OERiB7K0yrrtKVmM6LYvsmcGXdQvD-9JdvP54vv8Mr_69mUxP7_KFRdszDvBGmhbpWuhugIotpx3LVTKCK7byvASeWmg0QWAMU2ra0SBVc27xjRlU7GT7ONedzO1ve6UHsaATm6C7TFspUcr__0Z7Fqu_C9ZAC8LzpLA-4NA8D8nHUfZ2_gQdNB-irJKx6-bZPZ_IBUJK4Em8HQPquBjDNr8cUNB7lqVqVX50KqEOuGv_07wCB9qTMC7A4AxVWcCDsrGR06AYDXd7X2759Z2tb6zQUuMvbxVveSlFLIQxS7tmz1j0EtchaRzs0x3Z8kOcCYY-w0PWsH6</recordid><startdate>20070901</startdate><enddate>20070901</enddate><creator>Marianelli, Cinzia</creator><creator>Graziani, Caterina</creator><creator>Santangelo, Carmela</creator><creator>Xibilia, Maria Teresa</creator><creator>Imbriani, Alida</creator><creator>Amato, Rosa</creator><creator>Neri, Domenico</creator><creator>Cuccia, Mario</creator><creator>Rinnone, Sebastiano</creator><creator>Di Marco, Vincenzo</creator><creator>Ciuchini, Franco</creator><general>American Society for Microbiology</general><scope>FBQ</scope><scope>IQODW</scope><scope>CGR</scope><scope>CUY</scope><scope>CVF</scope><scope>ECM</scope><scope>EIF</scope><scope>NPM</scope><scope>AAYXX</scope><scope>CITATION</scope><scope>7QL</scope><scope>7TM</scope><scope>C1K</scope><scope>7X8</scope><scope>5PM</scope></search><sort><creationdate>20070901</creationdate><title>Molecular Epidemiological and Antibiotic Susceptibility Characterization of Brucella Isolates from Humans in Sicily, Italy</title><author>Marianelli, Cinzia ; Graziani, Caterina ; Santangelo, Carmela ; Xibilia, Maria Teresa ; Imbriani, Alida ; Amato, Rosa ; Neri, Domenico ; Cuccia, Mario ; Rinnone, Sebastiano ; Di Marco, Vincenzo ; Ciuchini, Franco</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c493t-d9380bbce79cd201ab44db06cf94eb6f45a45f08e200ff8be7aa9a674d8f85863</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2007</creationdate><topic>Anti-Bacterial Agents - pharmacology</topic><topic>Bacterial Proteins - genetics</topic><topic>Bacterial Typing Techniques</topic><topic>Bacteriology</topic><topic>Biological and medical sciences</topic><topic>Brucella melitensis</topic><topic>Brucella melitensis - classification</topic><topic>Brucella melitensis - drug effects</topic><topic>Brucella melitensis - genetics</topic><topic>Brucella melitensis - isolation & purification</topic><topic>Brucellosis - epidemiology</topic><topic>Brucellosis - microbiology</topic><topic>DNA, Bacterial - genetics</topic><topic>DNA-Directed RNA Polymerases - genetics</topic><topic>Epidemiology</topic><topic>Fundamental and applied biological sciences. Psychology</topic><topic>Genotype</topic><topic>Humans</topic><topic>Microbial Sensitivity Tests - methods</topic><topic>Microbiology</topic><topic>Miscellaneous</topic><topic>Molecular Epidemiology</topic><topic>Polymorphism, Genetic</topic><topic>Polymorphism, Restriction Fragment Length</topic><topic>Porins - genetics</topic><topic>Sequence Analysis, DNA</topic><topic>Sicily - epidemiology</topic><topic>Tandem Repeat Sequences - genetics</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Marianelli, Cinzia</creatorcontrib><creatorcontrib>Graziani, Caterina</creatorcontrib><creatorcontrib>Santangelo, Carmela</creatorcontrib><creatorcontrib>Xibilia, Maria Teresa</creatorcontrib><creatorcontrib>Imbriani, Alida</creatorcontrib><creatorcontrib>Amato, Rosa</creatorcontrib><creatorcontrib>Neri, Domenico</creatorcontrib><creatorcontrib>Cuccia, Mario</creatorcontrib><creatorcontrib>Rinnone, Sebastiano</creatorcontrib><creatorcontrib>Di Marco, Vincenzo</creatorcontrib><creatorcontrib>Ciuchini, Franco</creatorcontrib><collection>AGRIS</collection><collection>Pascal-Francis</collection><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>Bacteriology Abstracts (Microbiology B)</collection><collection>Nucleic Acids Abstracts</collection><collection>Environmental Sciences and Pollution Management</collection><collection>MEDLINE - Academic</collection><collection>PubMed Central (Full Participant titles)</collection><jtitle>Journal of Clinical Microbiology</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Marianelli, Cinzia</au><au>Graziani, Caterina</au><au>Santangelo, Carmela</au><au>Xibilia, Maria Teresa</au><au>Imbriani, Alida</au><au>Amato, Rosa</au><au>Neri, Domenico</au><au>Cuccia, Mario</au><au>Rinnone, Sebastiano</au><au>Di Marco, Vincenzo</au><au>Ciuchini, Franco</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Molecular Epidemiological and Antibiotic Susceptibility Characterization of Brucella Isolates from Humans in Sicily, Italy</atitle><jtitle>Journal of Clinical Microbiology</jtitle><addtitle>J Clin Microbiol</addtitle><date>2007-09-01</date><risdate>2007</risdate><volume>45</volume><issue>9</issue><spage>2923</spage><epage>2928</epage><pages>2923-2928</pages><issn>0095-1137</issn><eissn>1098-660X</eissn><eissn>1098-5530</eissn><coden>JCMIDW</coden><abstract>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.</abstract><cop>Washington, DC</cop><pub>American Society for Microbiology</pub><pmid>17634297</pmid><doi>10.1128/JCM.00822-07</doi><tpages>6</tpages><oa>free_for_read</oa></addata></record> |
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subjects | Anti-Bacterial Agents - pharmacology Bacterial Proteins - genetics Bacterial Typing Techniques Bacteriology Biological and medical sciences Brucella melitensis Brucella melitensis - classification Brucella melitensis - drug effects Brucella melitensis - genetics Brucella melitensis - isolation & purification Brucellosis - epidemiology Brucellosis - microbiology DNA, Bacterial - genetics DNA-Directed RNA Polymerases - genetics Epidemiology Fundamental and applied biological sciences. Psychology Genotype Humans Microbial Sensitivity Tests - methods Microbiology Miscellaneous Molecular Epidemiology Polymorphism, Genetic Polymorphism, Restriction Fragment Length Porins - genetics Sequence Analysis, DNA Sicily - epidemiology Tandem Repeat Sequences - genetics |
title | Molecular Epidemiological and Antibiotic Susceptibility Characterization of Brucella Isolates from Humans in Sicily, Italy |
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