Development and application of a triplex‐PCR assay for rapid detection of methicillin‐resistant Staphylococcus aureus from pigs
A triplex‐PCR assay was developed and evaluated for rapid detection of methicillin‐resistant Staphylococcus aureus (MRSA) recovered from various biological samples of pig. Three sets of primers were designed to target mecA, 16S rRNA and nuc genes of MRSA. The specific amplification generated three b...
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| Veröffentlicht in: | Letters in applied microbiology 2021-02, Vol.72 (2), p.121-125 |
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| creator | Rajkhowa, S. Pegu, S.R. Patil, G.P. Agrawal, R.K. |
| description | A triplex‐PCR assay was developed and evaluated for rapid detection of methicillin‐resistant Staphylococcus aureus (MRSA) recovered from various biological samples of pig. Three sets of primers were designed to target mecA, 16S rRNA and nuc genes of MRSA. The specific amplification generated three bands on agarose gel, with sizes 280 bp for mecA, 654 bp for 16S rRNA and 481 bp for nuc, respectively. A potential advantage of the PCR assay is its sensitivity with a detection limit of 102 CFU per ml of bacteria. In all, 79 MRSA isolates recovered from various samples of pigs were subjected to the amplification by the triplex‐PCR assay and all the isolates yielded three bands corresponding to the three genes under this study. No false‐positive amplification was observed, indicating the high specificity of the developed triplex‐PCR assay. This assay will be a useful and powerful method for differentiation of MRSA from methicillin‐sensitive S. aureus, coagulase‐negative methicillin‐resistant staphylococci and coagulase‐negative methicillin‐sensitive staphylococci.
Significance and Impact of the Study: The study reports the development and evaluation of a triplex‐PCR assay for the rapid detection of methicillin‐resistant Staphylococcus aureus (MRSA) from pigs. There is an urgent public health need for early and reliable detection of MRSA infection not only to reduce the cost of treatment but also to direct appropriate antibiotic therapy for its better management. This assay will be very useful for the rapid detection of MRSA from pigs. |
| doi_str_mv | 10.1111/lam.13408 |
| format | Article |
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Significance and Impact of the Study: The study reports the development and evaluation of a triplex‐PCR assay for the rapid detection of methicillin‐resistant Staphylococcus aureus (MRSA) from pigs. There is an urgent public health need for early and reliable detection of MRSA infection not only to reduce the cost of treatment but also to direct appropriate antibiotic therapy for its better management. This assay will be very useful for the rapid detection of MRSA from pigs.</description><identifier>ISSN: 0266-8254</identifier><identifier>EISSN: 1472-765X</identifier><identifier>DOI: 10.1111/lam.13408</identifier><identifier>PMID: 33090539</identifier><language>eng</language><publisher>England: Oxford University Press</publisher><subject>Amplification ; Animals ; Assaying ; Bacterial Proteins - genetics ; Biological properties ; Biological samples ; Coagulase ; DNA Primers - genetics ; Drug resistance ; Genes ; Humans ; Limit of Detection ; Methicillin ; Methicillin - pharmacology ; Methicillin Resistance - genetics ; Methicillin-Resistant Staphylococcus aureus - genetics ; Methicillin-Resistant Staphylococcus aureus - isolation & purification ; methicillin‐resistant Staphylococcus aureus ; Micrococcal Nuclease - genetics ; Multiplex Polymerase Chain Reaction - methods ; Penicillin-Binding Proteins - genetics ; pig ; RNA, Ribosomal, 16S - genetics ; rRNA 16S ; sensitivity ; specificity ; Staphylococcal Infections - diagnosis ; Staphylococcal Infections - microbiology ; Staphylococcal Infections - veterinary ; Staphylococcus aureus ; Staphylococcus infections ; Swine ; triplex PCR</subject><ispartof>Letters in applied microbiology, 2021-02, Vol.72 (2), p.121-125</ispartof><rights>2020 The Society for Applied Microbiology</rights><rights>2020 The Society for Applied Microbiology.</rights><rights>Copyright © 2021 The Society for Applied Microbiology</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c3538-d2989813f7e6d1a740ae6681066e2876f85872164f411347ceaa312fb26783f3</citedby><cites>FETCH-LOGICAL-c3538-d2989813f7e6d1a740ae6681066e2876f85872164f411347ceaa312fb26783f3</cites><orcidid>0000-0001-6296-1286</orcidid></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://onlinelibrary.wiley.com/doi/pdf/10.1111%2Flam.13408$$EPDF$$P50$$Gwiley$$H</linktopdf><linktohtml>$$Uhttps://onlinelibrary.wiley.com/doi/full/10.1111%2Flam.13408$$EHTML$$P50$$Gwiley$$H</linktohtml><link.rule.ids>314,780,784,1416,27923,27924,45573,45574</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/33090539$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Rajkhowa, S.</creatorcontrib><creatorcontrib>Pegu, S.R.</creatorcontrib><creatorcontrib>Patil, G.P.</creatorcontrib><creatorcontrib>Agrawal, R.K.</creatorcontrib><title>Development and application of a triplex‐PCR assay for rapid detection of methicillin‐resistant Staphylococcus aureus from pigs</title><title>Letters in applied microbiology</title><addtitle>Lett Appl Microbiol</addtitle><description>A triplex‐PCR assay was developed and evaluated for rapid detection of methicillin‐resistant Staphylococcus aureus (MRSA) recovered from various biological samples of pig. Three sets of primers were designed to target mecA, 16S rRNA and nuc genes of MRSA. The specific amplification generated three bands on agarose gel, with sizes 280 bp for mecA, 654 bp for 16S rRNA and 481 bp for nuc, respectively. A potential advantage of the PCR assay is its sensitivity with a detection limit of 102 CFU per ml of bacteria. In all, 79 MRSA isolates recovered from various samples of pigs were subjected to the amplification by the triplex‐PCR assay and all the isolates yielded three bands corresponding to the three genes under this study. No false‐positive amplification was observed, indicating the high specificity of the developed triplex‐PCR assay. This assay will be a useful and powerful method for differentiation of MRSA from methicillin‐sensitive S. aureus, coagulase‐negative methicillin‐resistant staphylococci and coagulase‐negative methicillin‐sensitive staphylococci.
Significance and Impact of the Study: The study reports the development and evaluation of a triplex‐PCR assay for the rapid detection of methicillin‐resistant Staphylococcus aureus (MRSA) from pigs. There is an urgent public health need for early and reliable detection of MRSA infection not only to reduce the cost of treatment but also to direct appropriate antibiotic therapy for its better management. This assay will be very useful for the rapid detection of MRSA from pigs.</description><subject>Amplification</subject><subject>Animals</subject><subject>Assaying</subject><subject>Bacterial Proteins - genetics</subject><subject>Biological properties</subject><subject>Biological samples</subject><subject>Coagulase</subject><subject>DNA Primers - genetics</subject><subject>Drug resistance</subject><subject>Genes</subject><subject>Humans</subject><subject>Limit of Detection</subject><subject>Methicillin</subject><subject>Methicillin - pharmacology</subject><subject>Methicillin Resistance - genetics</subject><subject>Methicillin-Resistant Staphylococcus aureus - genetics</subject><subject>Methicillin-Resistant Staphylococcus aureus - isolation & purification</subject><subject>methicillin‐resistant Staphylococcus aureus</subject><subject>Micrococcal Nuclease - genetics</subject><subject>Multiplex Polymerase Chain Reaction - methods</subject><subject>Penicillin-Binding Proteins - genetics</subject><subject>pig</subject><subject>RNA, Ribosomal, 16S - genetics</subject><subject>rRNA 16S</subject><subject>sensitivity</subject><subject>specificity</subject><subject>Staphylococcal Infections - diagnosis</subject><subject>Staphylococcal Infections - microbiology</subject><subject>Staphylococcal Infections - veterinary</subject><subject>Staphylococcus aureus</subject><subject>Staphylococcus infections</subject><subject>Swine</subject><subject>triplex PCR</subject><issn>0266-8254</issn><issn>1472-765X</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2021</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNp10c9u1DAQBnALgehSOPACyBIXOKT1n8R2jtW2BaRFIOiBWzR1xtSVEwc7od1bpb4Az8iTYNiWAxJzmctPn0bzEfKcswNe5jDAcMBlzcwDsuK1FpVWzZeHZMWEUpURTb1HnuR8yRgzXLSPyZ6UrGWNbFfk9hi_Y4jTgONMYewpTFPwFmYfRxodBTonPwW8_nnz4-P6E4WcYUtdTDTB5Hva44z2Hg84X3jrQ_Bj4QmzzzOU3M8zTBfbEG20dskUloRluRQHOvmv-Sl55CBkfHa398nZ6cnZ-m21-fDm3fpoU1nZSFP1ojWt4dJpVD0HXTNApQxnSqEwWjnTGC24ql3Nyze0RQDJhTsXShvp5D55tYudUvy2YJ67wWeLIcCIccmdqBupTCt1U-jLf-hlXNJYjivKcN4yVvOiXu-UTTHnhK6bkh8gbTvOut_FdKWY7k8xxb64S1zOB-z_yvsmCjjcgSsfcPv_pG5z9H4X-QuB15nf</recordid><startdate>202102</startdate><enddate>202102</enddate><creator>Rajkhowa, S.</creator><creator>Pegu, S.R.</creator><creator>Patil, G.P.</creator><creator>Agrawal, R.K.</creator><general>Oxford University Press</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>7QL</scope><scope>7QO</scope><scope>7ST</scope><scope>7T7</scope><scope>7TM</scope><scope>8FD</scope><scope>C1K</scope><scope>FR3</scope><scope>M7N</scope><scope>P64</scope><scope>SOI</scope><scope>7X8</scope><orcidid>https://orcid.org/0000-0001-6296-1286</orcidid></search><sort><creationdate>202102</creationdate><title>Development and application of a triplex‐PCR assay for rapid detection of methicillin‐resistant Staphylococcus aureus from pigs</title><author>Rajkhowa, S. ; Pegu, S.R. ; Patil, G.P. ; Agrawal, R.K.</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c3538-d2989813f7e6d1a740ae6681066e2876f85872164f411347ceaa312fb26783f3</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2021</creationdate><topic>Amplification</topic><topic>Animals</topic><topic>Assaying</topic><topic>Bacterial Proteins - genetics</topic><topic>Biological properties</topic><topic>Biological samples</topic><topic>Coagulase</topic><topic>DNA Primers - genetics</topic><topic>Drug resistance</topic><topic>Genes</topic><topic>Humans</topic><topic>Limit of Detection</topic><topic>Methicillin</topic><topic>Methicillin - pharmacology</topic><topic>Methicillin Resistance - genetics</topic><topic>Methicillin-Resistant Staphylococcus aureus - genetics</topic><topic>Methicillin-Resistant Staphylococcus aureus - isolation & purification</topic><topic>methicillin‐resistant Staphylococcus aureus</topic><topic>Micrococcal Nuclease - genetics</topic><topic>Multiplex Polymerase Chain Reaction - methods</topic><topic>Penicillin-Binding Proteins - genetics</topic><topic>pig</topic><topic>RNA, Ribosomal, 16S - genetics</topic><topic>rRNA 16S</topic><topic>sensitivity</topic><topic>specificity</topic><topic>Staphylococcal Infections - diagnosis</topic><topic>Staphylococcal Infections - microbiology</topic><topic>Staphylococcal Infections - veterinary</topic><topic>Staphylococcus aureus</topic><topic>Staphylococcus infections</topic><topic>Swine</topic><topic>triplex PCR</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Rajkhowa, S.</creatorcontrib><creatorcontrib>Pegu, S.R.</creatorcontrib><creatorcontrib>Patil, G.P.</creatorcontrib><creatorcontrib>Agrawal, R.K.</creatorcontrib><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>Biotechnology Research Abstracts</collection><collection>Environment Abstracts</collection><collection>Industrial and Applied Microbiology Abstracts (Microbiology A)</collection><collection>Nucleic Acids Abstracts</collection><collection>Technology Research Database</collection><collection>Environmental Sciences and Pollution Management</collection><collection>Engineering Research Database</collection><collection>Algology Mycology and Protozoology Abstracts (Microbiology C)</collection><collection>Biotechnology and BioEngineering Abstracts</collection><collection>Environment Abstracts</collection><collection>MEDLINE - Academic</collection><jtitle>Letters in applied microbiology</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Rajkhowa, S.</au><au>Pegu, S.R.</au><au>Patil, G.P.</au><au>Agrawal, R.K.</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Development and application of a triplex‐PCR assay for rapid detection of methicillin‐resistant Staphylococcus aureus from pigs</atitle><jtitle>Letters in applied microbiology</jtitle><addtitle>Lett Appl Microbiol</addtitle><date>2021-02</date><risdate>2021</risdate><volume>72</volume><issue>2</issue><spage>121</spage><epage>125</epage><pages>121-125</pages><issn>0266-8254</issn><eissn>1472-765X</eissn><abstract>A triplex‐PCR assay was developed and evaluated for rapid detection of methicillin‐resistant Staphylococcus aureus (MRSA) recovered from various biological samples of pig. Three sets of primers were designed to target mecA, 16S rRNA and nuc genes of MRSA. The specific amplification generated three bands on agarose gel, with sizes 280 bp for mecA, 654 bp for 16S rRNA and 481 bp for nuc, respectively. A potential advantage of the PCR assay is its sensitivity with a detection limit of 102 CFU per ml of bacteria. In all, 79 MRSA isolates recovered from various samples of pigs were subjected to the amplification by the triplex‐PCR assay and all the isolates yielded three bands corresponding to the three genes under this study. No false‐positive amplification was observed, indicating the high specificity of the developed triplex‐PCR assay. This assay will be a useful and powerful method for differentiation of MRSA from methicillin‐sensitive S. aureus, coagulase‐negative methicillin‐resistant staphylococci and coagulase‐negative methicillin‐sensitive staphylococci.
Significance and Impact of the Study: The study reports the development and evaluation of a triplex‐PCR assay for the rapid detection of methicillin‐resistant Staphylococcus aureus (MRSA) from pigs. There is an urgent public health need for early and reliable detection of MRSA infection not only to reduce the cost of treatment but also to direct appropriate antibiotic therapy for its better management. This assay will be very useful for the rapid detection of MRSA from pigs.</abstract><cop>England</cop><pub>Oxford University Press</pub><pmid>33090539</pmid><doi>10.1111/lam.13408</doi><tpages>5</tpages><orcidid>https://orcid.org/0000-0001-6296-1286</orcidid></addata></record> |
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| ispartof | Letters in applied microbiology, 2021-02, Vol.72 (2), p.121-125 |
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| source | MEDLINE; Oxford University Press Journals All Titles (1996-Current); EZB-FREE-00999 freely available EZB journals; Wiley Online Library All Journals; Alma/SFX Local Collection |
| subjects | Amplification Animals Assaying Bacterial Proteins - genetics Biological properties Biological samples Coagulase DNA Primers - genetics Drug resistance Genes Humans Limit of Detection Methicillin Methicillin - pharmacology Methicillin Resistance - genetics Methicillin-Resistant Staphylococcus aureus - genetics Methicillin-Resistant Staphylococcus aureus - isolation & purification methicillin‐resistant Staphylococcus aureus Micrococcal Nuclease - genetics Multiplex Polymerase Chain Reaction - methods Penicillin-Binding Proteins - genetics pig RNA, Ribosomal, 16S - genetics rRNA 16S sensitivity specificity Staphylococcal Infections - diagnosis Staphylococcal Infections - microbiology Staphylococcal Infections - veterinary Staphylococcus aureus Staphylococcus infections Swine triplex PCR |
| title | Development and application of a triplex‐PCR assay for rapid detection of methicillin‐resistant Staphylococcus aureus from pigs |
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