A sulfone group-labeled TEM-DNA probe: comparison with a 32P-labeled probe in dot-hybridization
A non-radioactive DNA probe for the TEM-type β-lactamase gene was obtained by using the ‘Chemoprobe’ system. It was used along with a 32P-labeled TEM probe to screen for TEM β-lactamase gene in 107 bacterial isolates representing 7 Gram-negative genera and previously classified as TEM-positive or ne...
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| Veröffentlicht in: | Journal of biochemical and biophysical methods 1988-08, Vol.16 (4), p.301-309 |
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| container_title | Journal of biochemical and biophysical methods |
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| creator | Jouvenot, M. Descotes, F. Remy-Martin, A. Adessi, G.L. |
| description | A non-radioactive DNA probe for the TEM-type β-lactamase gene was obtained by using the ‘Chemoprobe’ system. It was used along with a
32P-labeled TEM probe to screen for TEM β-lactamase gene in 107 bacterial isolates representing 7 Gram-negative genera and previously classified as TEM-positive or negative. The DNA to be tested was extracted from these bacterial isolates by the Birnboim-Doly method and, after blotting charged bylon membranes, it was submitted to hybridization with either the TEM ‘Chemoprobe’ or the
32P-TEM probe. The TEM ‘Chrmoprobe’ could be detect as few as 25 pg specific DNA if it was used at a concentration of 5 ng per cm
2 of membrane. The results obtained by both probes were concordant in 93.5% of the entire samples. The TEM ‘Chemoprobe’ was specific since only one false positive was observed. Furthermore, it appeared at least as sensitive as the
32P-labeled TEM probe. As the dot-hybridization with the sulfone-labeled probe was sensitive, simple ans easy to perform, it will be useful for large-scale screening in clinical laboratory. |
| doi_str_mv | 10.1016/0165-022X(88)90064-4 |
| format | Article |
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32P-labeled TEM probe to screen for TEM β-lactamase gene in 107 bacterial isolates representing 7 Gram-negative genera and previously classified as TEM-positive or negative. The DNA to be tested was extracted from these bacterial isolates by the Birnboim-Doly method and, after blotting charged bylon membranes, it was submitted to hybridization with either the TEM ‘Chemoprobe’ or the
32P-TEM probe. The TEM ‘Chrmoprobe’ could be detect as few as 25 pg specific DNA if it was used at a concentration of 5 ng per cm
2 of membrane. The results obtained by both probes were concordant in 93.5% of the entire samples. The TEM ‘Chemoprobe’ was specific since only one false positive was observed. Furthermore, it appeared at least as sensitive as the
32P-labeled TEM probe. As the dot-hybridization with the sulfone-labeled probe was sensitive, simple ans easy to perform, it will be useful for large-scale screening in clinical laboratory.</description><identifier>ISSN: 0165-022X</identifier><identifier>EISSN: 1872-857X</identifier><identifier>DOI: 10.1016/0165-022X(88)90064-4</identifier><identifier>PMID: 3065394</identifier><language>eng</language><publisher>Netherlands: Elsevier B.V</publisher><subject>beta-Lactamases - genetics ; DNA Probes ; Dot-hybridization ; Genes ; Genes, Bacterial ; Gram-negative bacteria ; Gram-Negative Bacteria - enzymology ; Gram-Negative Bacteria - genetics ; Nucleic Acid Hybridization ; Phosphorus Radioisotopes ; Plasmids ; Radioisotope Dilution Technique ; Sulfone-labeled DNA probe ; TEM-type β-lactmase gene</subject><ispartof>Journal of biochemical and biophysical methods, 1988-08, Vol.16 (4), p.301-309</ispartof><rights>1988</rights><woscitedreferencessubscribed>false</woscitedreferencessubscribed><cites>FETCH-LOGICAL-c1534-a33d215a45dd40c8a5f3d7997c78cd50e0a820f7c3427e07e2480c4b9bc689ae3</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><link.rule.ids>314,780,784,27924,27925</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/3065394$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Jouvenot, M.</creatorcontrib><creatorcontrib>Descotes, F.</creatorcontrib><creatorcontrib>Remy-Martin, A.</creatorcontrib><creatorcontrib>Adessi, G.L.</creatorcontrib><title>A sulfone group-labeled TEM-DNA probe: comparison with a 32P-labeled probe in dot-hybridization</title><title>Journal of biochemical and biophysical methods</title><addtitle>J Biochem Biophys Methods</addtitle><description>A non-radioactive DNA probe for the TEM-type β-lactamase gene was obtained by using the ‘Chemoprobe’ system. It was used along with a
32P-labeled TEM probe to screen for TEM β-lactamase gene in 107 bacterial isolates representing 7 Gram-negative genera and previously classified as TEM-positive or negative. The DNA to be tested was extracted from these bacterial isolates by the Birnboim-Doly method and, after blotting charged bylon membranes, it was submitted to hybridization with either the TEM ‘Chemoprobe’ or the
32P-TEM probe. The TEM ‘Chrmoprobe’ could be detect as few as 25 pg specific DNA if it was used at a concentration of 5 ng per cm
2 of membrane. The results obtained by both probes were concordant in 93.5% of the entire samples. The TEM ‘Chemoprobe’ was specific since only one false positive was observed. Furthermore, it appeared at least as sensitive as the
32P-labeled TEM probe. As the dot-hybridization with the sulfone-labeled probe was sensitive, simple ans easy to perform, it will be useful for large-scale screening in clinical laboratory.</description><subject>beta-Lactamases - genetics</subject><subject>DNA Probes</subject><subject>Dot-hybridization</subject><subject>Genes</subject><subject>Genes, Bacterial</subject><subject>Gram-negative bacteria</subject><subject>Gram-Negative Bacteria - enzymology</subject><subject>Gram-Negative Bacteria - genetics</subject><subject>Nucleic Acid Hybridization</subject><subject>Phosphorus Radioisotopes</subject><subject>Plasmids</subject><subject>Radioisotope Dilution Technique</subject><subject>Sulfone-labeled DNA probe</subject><subject>TEM-type β-lactmase gene</subject><issn>0165-022X</issn><issn>1872-857X</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>1988</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNp9kDlv3DAQhYkghrNe5x8kAKvALmhTPEQqRYCFb8BXsQHcERQ5ihloxQ0p2bB_vbUHtkwxmOK9mTfzIfStoCcFLcrTsSShjD0daX1cUVoKIj6hSaEVI1qqp89osrN8QQc5_6WUcs3EPtrntJS8EhNkZjgPbRM7wH9SHJaktTW04PH84o6c38_wMsUafmIXF0ubQo4dfg39M7aYs8edeW3CocM-9uT5rU7Bh3fbh9gdor3Gthm-bvsU_b68mJ9dk9uHq5uz2S1xheSCWM49K6QV0ntBnbay4V5VlXJKOy8pUKsZbZTjgimgCpjQ1Im6ql2pKwt8in5s9o6n_Bsg92YRsoO2tR3EIRulSynlmDVFYmN0KeacoDHLFBY2vZmCmhVXs4JmVtCM1mbN1Yhx7Pt2_1AvwO-GtiBH_ddGh_HJlwDJZBegc-BDAtcbH8P_Az4AhSGGaQ</recordid><startdate>198808</startdate><enddate>198808</enddate><creator>Jouvenot, M.</creator><creator>Descotes, F.</creator><creator>Remy-Martin, A.</creator><creator>Adessi, G.L.</creator><general>Elsevier B.V</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></search><sort><creationdate>198808</creationdate><title>A sulfone group-labeled TEM-DNA probe: comparison with a 32P-labeled probe in dot-hybridization</title><author>Jouvenot, M. ; Descotes, F. ; Remy-Martin, A. ; Adessi, G.L.</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c1534-a33d215a45dd40c8a5f3d7997c78cd50e0a820f7c3427e07e2480c4b9bc689ae3</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>1988</creationdate><topic>beta-Lactamases - genetics</topic><topic>DNA Probes</topic><topic>Dot-hybridization</topic><topic>Genes</topic><topic>Genes, Bacterial</topic><topic>Gram-negative bacteria</topic><topic>Gram-Negative Bacteria - enzymology</topic><topic>Gram-Negative Bacteria - genetics</topic><topic>Nucleic Acid Hybridization</topic><topic>Phosphorus Radioisotopes</topic><topic>Plasmids</topic><topic>Radioisotope Dilution Technique</topic><topic>Sulfone-labeled DNA probe</topic><topic>TEM-type β-lactmase gene</topic><toplevel>online_resources</toplevel><creatorcontrib>Jouvenot, M.</creatorcontrib><creatorcontrib>Descotes, F.</creatorcontrib><creatorcontrib>Remy-Martin, A.</creatorcontrib><creatorcontrib>Adessi, G.L.</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><jtitle>Journal of biochemical and biophysical methods</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Jouvenot, M.</au><au>Descotes, F.</au><au>Remy-Martin, A.</au><au>Adessi, G.L.</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>A sulfone group-labeled TEM-DNA probe: comparison with a 32P-labeled probe in dot-hybridization</atitle><jtitle>Journal of biochemical and biophysical methods</jtitle><addtitle>J Biochem Biophys Methods</addtitle><date>1988-08</date><risdate>1988</risdate><volume>16</volume><issue>4</issue><spage>301</spage><epage>309</epage><pages>301-309</pages><issn>0165-022X</issn><eissn>1872-857X</eissn><abstract>A non-radioactive DNA probe for the TEM-type β-lactamase gene was obtained by using the ‘Chemoprobe’ system. It was used along with a
32P-labeled TEM probe to screen for TEM β-lactamase gene in 107 bacterial isolates representing 7 Gram-negative genera and previously classified as TEM-positive or negative. The DNA to be tested was extracted from these bacterial isolates by the Birnboim-Doly method and, after blotting charged bylon membranes, it was submitted to hybridization with either the TEM ‘Chemoprobe’ or the
32P-TEM probe. The TEM ‘Chrmoprobe’ could be detect as few as 25 pg specific DNA if it was used at a concentration of 5 ng per cm
2 of membrane. The results obtained by both probes were concordant in 93.5% of the entire samples. The TEM ‘Chemoprobe’ was specific since only one false positive was observed. Furthermore, it appeared at least as sensitive as the
32P-labeled TEM probe. As the dot-hybridization with the sulfone-labeled probe was sensitive, simple ans easy to perform, it will be useful for large-scale screening in clinical laboratory.</abstract><cop>Netherlands</cop><pub>Elsevier B.V</pub><pmid>3065394</pmid><doi>10.1016/0165-022X(88)90064-4</doi><tpages>9</tpages></addata></record> |
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| ispartof | Journal of biochemical and biophysical methods, 1988-08, Vol.16 (4), p.301-309 |
| issn | 0165-022X 1872-857X |
| language | eng |
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| source | MEDLINE; Alma/SFX Local Collection |
| subjects | beta-Lactamases - genetics DNA Probes Dot-hybridization Genes Genes, Bacterial Gram-negative bacteria Gram-Negative Bacteria - enzymology Gram-Negative Bacteria - genetics Nucleic Acid Hybridization Phosphorus Radioisotopes Plasmids Radioisotope Dilution Technique Sulfone-labeled DNA probe TEM-type β-lactmase gene |
| title | A sulfone group-labeled TEM-DNA probe: comparison with a 32P-labeled probe in dot-hybridization |
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