Loop-mediated isothermal amplification assay targeting the omp25 gene for rapid detection of Brucella spp
A novel loop-mediated isothermal amplification (LAMP) assay was established to detect Brucella species DNA in milk and blood samples of animals and humans. This LAMP assay based on the sequence of highly repetitive omp25 gene was able to detect 9 fg/μl Brucella spp. DNA with high sensitivity, which...
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| Veröffentlicht in: | Molecular and cellular probes 2011-04, Vol.25 (2), p.126-129 |
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| container_title | Molecular and cellular probes |
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| creator | Lin, Guo-Zhen Zheng, Fu-Ying Zhou, Ji-Zhang Gong, Xiao-Wei Wang, Guang-Hua Cao, Xiao-An Qiu, Chang-Qing |
| description | A novel loop-mediated isothermal amplification (LAMP) assay was established to detect
Brucella species DNA in milk and blood samples of animals and humans. This LAMP assay based on the sequence of highly repetitive
omp25 gene was able to detect 9
fg/μl
Brucella spp. DNA with high sensitivity, which was 10 times higher than the nested PCR. The LAMP was evaluated for its specificity using 19 strains of six
Brucella species and 28 related non-
Brucella micro-organism strains as controls. The target 19
Brucella strains were all amplified, and no cross-reaction was found with all the non-
Brucella micro-organism strains. Both nested PCR and LAMP assays were then used to detect
Brucella spp. DNA in 78 milk samples and 113 blood samples from animals and 11 blood samples from humans, and the established LAMP assay yielded 99.0% concordance rate with the nested PCR. The LAMP assay should be a potential tool with high convenience, rapidity, sensitivity and specificity for the diagnosis of Brucellosis. |
| doi_str_mv | 10.1016/j.mcp.2011.01.001 |
| format | Article |
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Brucella species DNA in milk and blood samples of animals and humans. This LAMP assay based on the sequence of highly repetitive
omp25 gene was able to detect 9
fg/μl
Brucella spp. DNA with high sensitivity, which was 10 times higher than the nested PCR. The LAMP was evaluated for its specificity using 19 strains of six
Brucella species and 28 related non-
Brucella micro-organism strains as controls. The target 19
Brucella strains were all amplified, and no cross-reaction was found with all the non-
Brucella micro-organism strains. Both nested PCR and LAMP assays were then used to detect
Brucella spp. DNA in 78 milk samples and 113 blood samples from animals and 11 blood samples from humans, and the established LAMP assay yielded 99.0% concordance rate with the nested PCR. The LAMP assay should be a potential tool with high convenience, rapidity, sensitivity and specificity for the diagnosis of Brucellosis.</description><identifier>ISSN: 0890-8508</identifier><identifier>EISSN: 1096-1194</identifier><identifier>DOI: 10.1016/j.mcp.2011.01.001</identifier><identifier>PMID: 21232598</identifier><language>eng</language><publisher>England: Elsevier Ltd</publisher><subject>Animals ; Bacteria - classification ; Bacteria - genetics ; Bacterial Outer Membrane Proteins - genetics ; Brucella ; Brucella - classification ; Brucella - genetics ; Brucella spp ; Brucellosis - blood ; Brucellosis - diagnosis ; Brucellosis - microbiology ; Cattle ; DNA Primers - genetics ; DNA, Bacterial - blood ; DNA, Bacterial - genetics ; Female ; Humans ; LAMP ; Milk - microbiology ; Nucleic Acid Amplification Techniques - methods ; omp25 ; Polymerase Chain Reaction - methods ; Reproducibility of Results ; Sensitivity and Specificity ; Sheep ; Species Specificity ; Temperature</subject><ispartof>Molecular and cellular probes, 2011-04, Vol.25 (2), p.126-129</ispartof><rights>2011 Elsevier Ltd</rights><rights>Copyright © 2011 Elsevier Ltd. All rights reserved.</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c384t-d028485cf74b5749e3ff248b32b3f0a98553f263d8edc2391df9e2d8c0e0a003</citedby><cites>FETCH-LOGICAL-c384t-d028485cf74b5749e3ff248b32b3f0a98553f263d8edc2391df9e2d8c0e0a003</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktohtml>$$Uhttps://dx.doi.org/10.1016/j.mcp.2011.01.001$$EHTML$$P50$$Gelsevier$$H</linktohtml><link.rule.ids>314,777,781,3537,27905,27906,45976</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/21232598$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Lin, Guo-Zhen</creatorcontrib><creatorcontrib>Zheng, Fu-Ying</creatorcontrib><creatorcontrib>Zhou, Ji-Zhang</creatorcontrib><creatorcontrib>Gong, Xiao-Wei</creatorcontrib><creatorcontrib>Wang, Guang-Hua</creatorcontrib><creatorcontrib>Cao, Xiao-An</creatorcontrib><creatorcontrib>Qiu, Chang-Qing</creatorcontrib><title>Loop-mediated isothermal amplification assay targeting the omp25 gene for rapid detection of Brucella spp</title><title>Molecular and cellular probes</title><addtitle>Mol Cell Probes</addtitle><description>A novel loop-mediated isothermal amplification (LAMP) assay was established to detect
Brucella species DNA in milk and blood samples of animals and humans. This LAMP assay based on the sequence of highly repetitive
omp25 gene was able to detect 9
fg/μl
Brucella spp. DNA with high sensitivity, which was 10 times higher than the nested PCR. The LAMP was evaluated for its specificity using 19 strains of six
Brucella species and 28 related non-
Brucella micro-organism strains as controls. The target 19
Brucella strains were all amplified, and no cross-reaction was found with all the non-
Brucella micro-organism strains. Both nested PCR and LAMP assays were then used to detect
Brucella spp. DNA in 78 milk samples and 113 blood samples from animals and 11 blood samples from humans, and the established LAMP assay yielded 99.0% concordance rate with the nested PCR. The LAMP assay should be a potential tool with high convenience, rapidity, sensitivity and specificity for the diagnosis of Brucellosis.</description><subject>Animals</subject><subject>Bacteria - classification</subject><subject>Bacteria - genetics</subject><subject>Bacterial Outer Membrane Proteins - genetics</subject><subject>Brucella</subject><subject>Brucella - classification</subject><subject>Brucella - genetics</subject><subject>Brucella spp</subject><subject>Brucellosis - blood</subject><subject>Brucellosis - diagnosis</subject><subject>Brucellosis - microbiology</subject><subject>Cattle</subject><subject>DNA Primers - genetics</subject><subject>DNA, Bacterial - blood</subject><subject>DNA, Bacterial - genetics</subject><subject>Female</subject><subject>Humans</subject><subject>LAMP</subject><subject>Milk - microbiology</subject><subject>Nucleic Acid Amplification Techniques - methods</subject><subject>omp25</subject><subject>Polymerase Chain Reaction - methods</subject><subject>Reproducibility of Results</subject><subject>Sensitivity and Specificity</subject><subject>Sheep</subject><subject>Species Specificity</subject><subject>Temperature</subject><issn>0890-8508</issn><issn>1096-1194</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2011</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNqFkTFvFDEUhC1ERI6EH0CD3FHt8Wyvd21RQUQA6SSa9JbPfj582l0bew8p_x4nF1KCNNJrvhk9zRDylsGWARs-HLezy1sOjG2hCdgLsmGgh44x3b8kG1AaOiVBXZLXtR4BQPegXpFLzrjgUqsNibuUcjejj3ZFT2NN608ss52onfMUQ3R2jWmhtlZ7T1dbDrjG5UAbRdOcuaQHXJCGVGixOXrqcUX3aEmBfi4nh9Nkac35mlwEO1V883SvyN3tl7ubb93ux9fvN592nROqXzsPXPVKujD2ezn2GkUIvFd7wfcigNVKShH4ILxC77jQzAeN3CsHCBZAXJH359hc0q8T1tXMsT4-sWA6VaNhZHIYh_6_pBr6UclxVI1kZ9KVVGvBYHKJsy33hoF5WMIcTVvCPCxhoAlY87x7Sj_tW73Pjr_VN-DjGcBWxu-IxVQXcXFtitIaND7Ff8T_AXovmZg</recordid><startdate>20110401</startdate><enddate>20110401</enddate><creator>Lin, Guo-Zhen</creator><creator>Zheng, Fu-Ying</creator><creator>Zhou, Ji-Zhang</creator><creator>Gong, Xiao-Wei</creator><creator>Wang, Guang-Hua</creator><creator>Cao, Xiao-An</creator><creator>Qiu, Chang-Qing</creator><general>Elsevier Ltd</general><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>7X8</scope><scope>7QL</scope><scope>C1K</scope></search><sort><creationdate>20110401</creationdate><title>Loop-mediated isothermal amplification assay targeting the omp25 gene for rapid detection of Brucella spp</title><author>Lin, Guo-Zhen ; Zheng, Fu-Ying ; Zhou, Ji-Zhang ; Gong, Xiao-Wei ; Wang, Guang-Hua ; Cao, Xiao-An ; Qiu, Chang-Qing</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c384t-d028485cf74b5749e3ff248b32b3f0a98553f263d8edc2391df9e2d8c0e0a003</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2011</creationdate><topic>Animals</topic><topic>Bacteria - classification</topic><topic>Bacteria - genetics</topic><topic>Bacterial Outer Membrane Proteins - genetics</topic><topic>Brucella</topic><topic>Brucella - classification</topic><topic>Brucella - genetics</topic><topic>Brucella spp</topic><topic>Brucellosis - blood</topic><topic>Brucellosis - diagnosis</topic><topic>Brucellosis - microbiology</topic><topic>Cattle</topic><topic>DNA Primers - genetics</topic><topic>DNA, Bacterial - blood</topic><topic>DNA, Bacterial - genetics</topic><topic>Female</topic><topic>Humans</topic><topic>LAMP</topic><topic>Milk - microbiology</topic><topic>Nucleic Acid Amplification Techniques - methods</topic><topic>omp25</topic><topic>Polymerase Chain Reaction - methods</topic><topic>Reproducibility of Results</topic><topic>Sensitivity and Specificity</topic><topic>Sheep</topic><topic>Species Specificity</topic><topic>Temperature</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Lin, Guo-Zhen</creatorcontrib><creatorcontrib>Zheng, Fu-Ying</creatorcontrib><creatorcontrib>Zhou, Ji-Zhang</creatorcontrib><creatorcontrib>Gong, Xiao-Wei</creatorcontrib><creatorcontrib>Wang, Guang-Hua</creatorcontrib><creatorcontrib>Cao, Xiao-An</creatorcontrib><creatorcontrib>Qiu, Chang-Qing</creatorcontrib><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>MEDLINE - Academic</collection><collection>Bacteriology Abstracts (Microbiology B)</collection><collection>Environmental Sciences and Pollution Management</collection><jtitle>Molecular and cellular probes</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Lin, Guo-Zhen</au><au>Zheng, Fu-Ying</au><au>Zhou, Ji-Zhang</au><au>Gong, Xiao-Wei</au><au>Wang, Guang-Hua</au><au>Cao, Xiao-An</au><au>Qiu, Chang-Qing</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Loop-mediated isothermal amplification assay targeting the omp25 gene for rapid detection of Brucella spp</atitle><jtitle>Molecular and cellular probes</jtitle><addtitle>Mol Cell Probes</addtitle><date>2011-04-01</date><risdate>2011</risdate><volume>25</volume><issue>2</issue><spage>126</spage><epage>129</epage><pages>126-129</pages><issn>0890-8508</issn><eissn>1096-1194</eissn><abstract>A novel loop-mediated isothermal amplification (LAMP) assay was established to detect
Brucella species DNA in milk and blood samples of animals and humans. This LAMP assay based on the sequence of highly repetitive
omp25 gene was able to detect 9
fg/μl
Brucella spp. DNA with high sensitivity, which was 10 times higher than the nested PCR. The LAMP was evaluated for its specificity using 19 strains of six
Brucella species and 28 related non-
Brucella micro-organism strains as controls. The target 19
Brucella strains were all amplified, and no cross-reaction was found with all the non-
Brucella micro-organism strains. Both nested PCR and LAMP assays were then used to detect
Brucella spp. DNA in 78 milk samples and 113 blood samples from animals and 11 blood samples from humans, and the established LAMP assay yielded 99.0% concordance rate with the nested PCR. The LAMP assay should be a potential tool with high convenience, rapidity, sensitivity and specificity for the diagnosis of Brucellosis.</abstract><cop>England</cop><pub>Elsevier Ltd</pub><pmid>21232598</pmid><doi>10.1016/j.mcp.2011.01.001</doi><tpages>4</tpages></addata></record> |
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| ispartof | Molecular and cellular probes, 2011-04, Vol.25 (2), p.126-129 |
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| language | eng |
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| source | MEDLINE; Elsevier ScienceDirect Journals |
| subjects | Animals Bacteria - classification Bacteria - genetics Bacterial Outer Membrane Proteins - genetics Brucella Brucella - classification Brucella - genetics Brucella spp Brucellosis - blood Brucellosis - diagnosis Brucellosis - microbiology Cattle DNA Primers - genetics DNA, Bacterial - blood DNA, Bacterial - genetics Female Humans LAMP Milk - microbiology Nucleic Acid Amplification Techniques - methods omp25 Polymerase Chain Reaction - methods Reproducibility of Results Sensitivity and Specificity Sheep Species Specificity Temperature |
| title | Loop-mediated isothermal amplification assay targeting the omp25 gene for rapid detection of Brucella spp |
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