The 7H11 Agar Medium Supplemented with Calf Bovine Serum for Susceptibility Testing of Mycobacterium tuberculosis Isolates Against Pyrazinamide
Despite its importance, pyrazinamide (PZA) is a blind spot in drug susceptibility testing in tuberculosis laboratories. The aim of this study was to set up a reliable agar-based proportion method for detection of PZA-resistant phenotypes using Middlebrook 7H11 agar supplemented with calf bovine seru...
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| Veröffentlicht in: | Microbial drug resistance (Larchmont, N.Y.) N.Y.), 2021-12, Vol.27 (12), p.1652-1657 |
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| container_title | Microbial drug resistance (Larchmont, N.Y.) |
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| creator | Amini, Sirus Kardan-Yamchi, Jalil Kazemian, Hossein Nasiri, Mohammad Javad Hamzehloo, Gholamreza Hoffner, Sven Feizabadi, Mohammad Mehdi |
| description | Despite its importance, pyrazinamide (PZA) is a blind spot in drug susceptibility testing in tuberculosis laboratories. The aim of this study was to set up a reliable agar-based proportion method for detection of PZA-resistant phenotypes using Middlebrook 7H11 agar supplemented with calf bovine serum (CBS) compared with albumin/dextrose/catalase (ADC) enrichment and
pncA
/
rpsA
sequencing results. The 7H11 agar medium supplemented with 10% ADC or 10% CBS (pH 6.2) and 100 μg/mL PZA was used to detect PZA resistance among 64
Mycobacterium tuberculosis
isolates. Sanger sequencing and whole-genome sequencing were performed to track mutations in the
pncA
,
rpsA,
and their upstream regions. A total of 43 rifampicin/multidrug-resistant, 20 drug-susceptible, and 1 isoniazid mono-resistant
M. tuberculosis
isolates were investigated. The 7H11+ADC and 7H11+CBS could detect 22 and 23 PZA-resistant strains, respectively. With the same specificity, the sensitivity and accuracy of 7H11+CBS was found to be a little greater than 7H11+ADC in PZA resistance detection compared with sequencing results. Twenty-four mutant strains were found to have different mutations in
pncA
-upstream,
pncA
and
rpsA
genes, in which Gly97Asp was the most dominant mutation. The results obtained from 7H11+CBS were comparable to the results of 7H11+ADC. Therefore, the 7H11 agar proportion method would be a less-expensive test using CBS and produces reliable results. |
| doi_str_mv | 10.1089/mdr.2020.0509 |
| format | Article |
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pncA
/
rpsA
sequencing results. The 7H11 agar medium supplemented with 10% ADC or 10% CBS (pH 6.2) and 100 μg/mL PZA was used to detect PZA resistance among 64
Mycobacterium tuberculosis
isolates. Sanger sequencing and whole-genome sequencing were performed to track mutations in the
pncA
,
rpsA,
and their upstream regions. A total of 43 rifampicin/multidrug-resistant, 20 drug-susceptible, and 1 isoniazid mono-resistant
M. tuberculosis
isolates were investigated. The 7H11+ADC and 7H11+CBS could detect 22 and 23 PZA-resistant strains, respectively. With the same specificity, the sensitivity and accuracy of 7H11+CBS was found to be a little greater than 7H11+ADC in PZA resistance detection compared with sequencing results. Twenty-four mutant strains were found to have different mutations in
pncA
-upstream,
pncA
and
rpsA
genes, in which Gly97Asp was the most dominant mutation. The results obtained from 7H11+CBS were comparable to the results of 7H11+ADC. Therefore, the 7H11 agar proportion method would be a less-expensive test using CBS and produces reliable results.</description><identifier>ISSN: 1076-6294</identifier><identifier>EISSN: 1931-8448</identifier><identifier>DOI: 10.1089/mdr.2020.0509</identifier><identifier>PMID: 34077245</identifier><language>eng</language><publisher>United States: Mary Ann Liebert, Inc., publishers</publisher><subject>Agar ; Albumins ; Amidohydrolases - genetics ; Antitubercular Agents - pharmacology ; Catalase ; Cattle ; Dextrose ; Drug Resistance, Bacterial - genetics ; Gene sequencing ; Genes, Bacterial ; Genomes ; Humans ; Isoniazid ; Mechanisms ; Microbial Sensitivity Tests - methods ; Multidrug resistance ; Mutation ; Mycobacterium tuberculosis ; Mycobacterium tuberculosis - drug effects ; Mycobacterium tuberculosis - genetics ; Phenotypes ; Pyrazinamide ; Pyrazinamide - pharmacology ; Rifampin ; Serum Albumin, Bovine ; Tuberculosis ; Whole Genome Sequencing</subject><ispartof>Microbial drug resistance (Larchmont, N.Y.), 2021-12, Vol.27 (12), p.1652-1657</ispartof><rights>2021, Mary Ann Liebert, Inc., publishers</rights><rights>Copyright Mary Ann Liebert, Inc. Dec 2021</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c403t-3fec52e5cbf97ac03e0bba9aef9a482cc0e949e3b492ca78bc3fceebd49793993</citedby><cites>FETCH-LOGICAL-c403t-3fec52e5cbf97ac03e0bba9aef9a482cc0e949e3b492ca78bc3fceebd49793993</cites><orcidid>0000-0001-8546-512X</orcidid></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><link.rule.ids>230,314,777,781,882,27905,27906</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/34077245$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink><backlink>$$Uhttp://kipublications.ki.se/Default.aspx?queryparsed=id:146810597$$DView record from Swedish Publication Index$$Hfree_for_read</backlink></links><search><creatorcontrib>Amini, Sirus</creatorcontrib><creatorcontrib>Kardan-Yamchi, Jalil</creatorcontrib><creatorcontrib>Kazemian, Hossein</creatorcontrib><creatorcontrib>Nasiri, Mohammad Javad</creatorcontrib><creatorcontrib>Hamzehloo, Gholamreza</creatorcontrib><creatorcontrib>Hoffner, Sven</creatorcontrib><creatorcontrib>Feizabadi, Mohammad Mehdi</creatorcontrib><title>The 7H11 Agar Medium Supplemented with Calf Bovine Serum for Susceptibility Testing of Mycobacterium tuberculosis Isolates Against Pyrazinamide</title><title>Microbial drug resistance (Larchmont, N.Y.)</title><addtitle>Microb Drug Resist</addtitle><description>Despite its importance, pyrazinamide (PZA) is a blind spot in drug susceptibility testing in tuberculosis laboratories. The aim of this study was to set up a reliable agar-based proportion method for detection of PZA-resistant phenotypes using Middlebrook 7H11 agar supplemented with calf bovine serum (CBS) compared with albumin/dextrose/catalase (ADC) enrichment and
pncA
/
rpsA
sequencing results. The 7H11 agar medium supplemented with 10% ADC or 10% CBS (pH 6.2) and 100 μg/mL PZA was used to detect PZA resistance among 64
Mycobacterium tuberculosis
isolates. Sanger sequencing and whole-genome sequencing were performed to track mutations in the
pncA
,
rpsA,
and their upstream regions. A total of 43 rifampicin/multidrug-resistant, 20 drug-susceptible, and 1 isoniazid mono-resistant
M. tuberculosis
isolates were investigated. The 7H11+ADC and 7H11+CBS could detect 22 and 23 PZA-resistant strains, respectively. With the same specificity, the sensitivity and accuracy of 7H11+CBS was found to be a little greater than 7H11+ADC in PZA resistance detection compared with sequencing results. Twenty-four mutant strains were found to have different mutations in
pncA
-upstream,
pncA
and
rpsA
genes, in which Gly97Asp was the most dominant mutation. The results obtained from 7H11+CBS were comparable to the results of 7H11+ADC. Therefore, the 7H11 agar proportion method would be a less-expensive test using CBS and produces reliable results.</description><subject>Agar</subject><subject>Albumins</subject><subject>Amidohydrolases - genetics</subject><subject>Antitubercular Agents - pharmacology</subject><subject>Catalase</subject><subject>Cattle</subject><subject>Dextrose</subject><subject>Drug Resistance, Bacterial - genetics</subject><subject>Gene sequencing</subject><subject>Genes, Bacterial</subject><subject>Genomes</subject><subject>Humans</subject><subject>Isoniazid</subject><subject>Mechanisms</subject><subject>Microbial Sensitivity Tests - methods</subject><subject>Multidrug resistance</subject><subject>Mutation</subject><subject>Mycobacterium tuberculosis</subject><subject>Mycobacterium tuberculosis - drug effects</subject><subject>Mycobacterium tuberculosis - genetics</subject><subject>Phenotypes</subject><subject>Pyrazinamide</subject><subject>Pyrazinamide - pharmacology</subject><subject>Rifampin</subject><subject>Serum Albumin, Bovine</subject><subject>Tuberculosis</subject><subject>Whole Genome Sequencing</subject><issn>1076-6294</issn><issn>1931-8448</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2021</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNqFkU1v1DAQhiMEoqVw5IosceGSxY6dDx_Lin5IrVqpy9mynXHr4sTBdlpt_0T_Mo52KRIXTjMaPXpn3nmL4iPBK4I7_nXow6rCFV7hGvNXxSHhlJQdY93r3OO2KZuKs4PiXYz3GOOaNPRtcUAZbtuK1YfF8-YOUHtGCDq-lQFdQm_nAd3M0-RggDFBjx5tukNr6Qz65h_sCOgGQmaMD5mLGqZklXU2bdEGYrLjLfIGXW61V1InCItemhUEPTsfbUTn0TuZIC4b7RgTut4G-WRHOdge3hdvjHQRPuzrUfHj5PtmfVZeXJ2er48vSs0wTSU1oOsKaq0Mb6XGFLBSkkswXLKu0hoDZxyoYrzSsu2UpkYDqJ7xllPO6VFR7nTjI0yzElOwgwxb4aUV-9HP3IFgDalrmvkvO34K_tecfYrBZu_OyRH8HEVV06bDhHWL9Od_0Hs_hzG7EVWD24Vj9O8BOvgYA5iXEwgWS7IiJyuWZMWSbOY_7VVnNUD_Qv-JMgN0ByxjOY7OQn56-o_sb6M8s1o</recordid><startdate>20211201</startdate><enddate>20211201</enddate><creator>Amini, Sirus</creator><creator>Kardan-Yamchi, Jalil</creator><creator>Kazemian, Hossein</creator><creator>Nasiri, Mohammad Javad</creator><creator>Hamzehloo, Gholamreza</creator><creator>Hoffner, Sven</creator><creator>Feizabadi, Mohammad Mehdi</creator><general>Mary Ann Liebert, Inc., publishers</general><general>Mary Ann Liebert, Inc</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>7QP</scope><scope>7QR</scope><scope>7T7</scope><scope>7TK</scope><scope>8FD</scope><scope>C1K</scope><scope>FR3</scope><scope>K9.</scope><scope>M7N</scope><scope>P64</scope><scope>7X8</scope><scope>ADTPV</scope><scope>AOWAS</scope><orcidid>https://orcid.org/0000-0001-8546-512X</orcidid></search><sort><creationdate>20211201</creationdate><title>The 7H11 Agar Medium Supplemented with Calf Bovine Serum for Susceptibility Testing of Mycobacterium tuberculosis Isolates Against Pyrazinamide</title><author>Amini, Sirus ; Kardan-Yamchi, Jalil ; Kazemian, Hossein ; Nasiri, Mohammad Javad ; Hamzehloo, Gholamreza ; Hoffner, Sven ; Feizabadi, Mohammad Mehdi</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c403t-3fec52e5cbf97ac03e0bba9aef9a482cc0e949e3b492ca78bc3fceebd49793993</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2021</creationdate><topic>Agar</topic><topic>Albumins</topic><topic>Amidohydrolases - genetics</topic><topic>Antitubercular Agents - pharmacology</topic><topic>Catalase</topic><topic>Cattle</topic><topic>Dextrose</topic><topic>Drug Resistance, Bacterial - genetics</topic><topic>Gene sequencing</topic><topic>Genes, Bacterial</topic><topic>Genomes</topic><topic>Humans</topic><topic>Isoniazid</topic><topic>Mechanisms</topic><topic>Microbial Sensitivity Tests - methods</topic><topic>Multidrug resistance</topic><topic>Mutation</topic><topic>Mycobacterium tuberculosis</topic><topic>Mycobacterium tuberculosis - drug effects</topic><topic>Mycobacterium tuberculosis - genetics</topic><topic>Phenotypes</topic><topic>Pyrazinamide</topic><topic>Pyrazinamide - pharmacology</topic><topic>Rifampin</topic><topic>Serum Albumin, Bovine</topic><topic>Tuberculosis</topic><topic>Whole Genome Sequencing</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Amini, Sirus</creatorcontrib><creatorcontrib>Kardan-Yamchi, Jalil</creatorcontrib><creatorcontrib>Kazemian, Hossein</creatorcontrib><creatorcontrib>Nasiri, Mohammad Javad</creatorcontrib><creatorcontrib>Hamzehloo, Gholamreza</creatorcontrib><creatorcontrib>Hoffner, Sven</creatorcontrib><creatorcontrib>Feizabadi, Mohammad Mehdi</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>Calcium & Calcified Tissue Abstracts</collection><collection>Chemoreception Abstracts</collection><collection>Industrial and Applied Microbiology Abstracts (Microbiology A)</collection><collection>Neurosciences Abstracts</collection><collection>Technology Research Database</collection><collection>Environmental Sciences and Pollution Management</collection><collection>Engineering Research Database</collection><collection>ProQuest Health & Medical Complete (Alumni)</collection><collection>Algology Mycology and Protozoology Abstracts (Microbiology C)</collection><collection>Biotechnology and BioEngineering Abstracts</collection><collection>MEDLINE - Academic</collection><collection>SwePub</collection><collection>SwePub Articles</collection><jtitle>Microbial drug resistance (Larchmont, N.Y.)</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Amini, Sirus</au><au>Kardan-Yamchi, Jalil</au><au>Kazemian, Hossein</au><au>Nasiri, Mohammad Javad</au><au>Hamzehloo, Gholamreza</au><au>Hoffner, Sven</au><au>Feizabadi, Mohammad Mehdi</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>The 7H11 Agar Medium Supplemented with Calf Bovine Serum for Susceptibility Testing of Mycobacterium tuberculosis Isolates Against Pyrazinamide</atitle><jtitle>Microbial drug resistance (Larchmont, N.Y.)</jtitle><addtitle>Microb Drug Resist</addtitle><date>2021-12-01</date><risdate>2021</risdate><volume>27</volume><issue>12</issue><spage>1652</spage><epage>1657</epage><pages>1652-1657</pages><issn>1076-6294</issn><eissn>1931-8448</eissn><abstract>Despite its importance, pyrazinamide (PZA) is a blind spot in drug susceptibility testing in tuberculosis laboratories. The aim of this study was to set up a reliable agar-based proportion method for detection of PZA-resistant phenotypes using Middlebrook 7H11 agar supplemented with calf bovine serum (CBS) compared with albumin/dextrose/catalase (ADC) enrichment and
pncA
/
rpsA
sequencing results. The 7H11 agar medium supplemented with 10% ADC or 10% CBS (pH 6.2) and 100 μg/mL PZA was used to detect PZA resistance among 64
Mycobacterium tuberculosis
isolates. Sanger sequencing and whole-genome sequencing were performed to track mutations in the
pncA
,
rpsA,
and their upstream regions. A total of 43 rifampicin/multidrug-resistant, 20 drug-susceptible, and 1 isoniazid mono-resistant
M. tuberculosis
isolates were investigated. The 7H11+ADC and 7H11+CBS could detect 22 and 23 PZA-resistant strains, respectively. With the same specificity, the sensitivity and accuracy of 7H11+CBS was found to be a little greater than 7H11+ADC in PZA resistance detection compared with sequencing results. Twenty-four mutant strains were found to have different mutations in
pncA
-upstream,
pncA
and
rpsA
genes, in which Gly97Asp was the most dominant mutation. The results obtained from 7H11+CBS were comparable to the results of 7H11+ADC. Therefore, the 7H11 agar proportion method would be a less-expensive test using CBS and produces reliable results.</abstract><cop>United States</cop><pub>Mary Ann Liebert, Inc., publishers</pub><pmid>34077245</pmid><doi>10.1089/mdr.2020.0509</doi><tpages>6</tpages><orcidid>https://orcid.org/0000-0001-8546-512X</orcidid></addata></record> |
| fulltext | fulltext |
| identifier | ISSN: 1076-6294 |
| ispartof | Microbial drug resistance (Larchmont, N.Y.), 2021-12, Vol.27 (12), p.1652-1657 |
| issn | 1076-6294 1931-8448 |
| language | eng |
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| source | MEDLINE; Alma/SFX Local Collection |
| subjects | Agar Albumins Amidohydrolases - genetics Antitubercular Agents - pharmacology Catalase Cattle Dextrose Drug Resistance, Bacterial - genetics Gene sequencing Genes, Bacterial Genomes Humans Isoniazid Mechanisms Microbial Sensitivity Tests - methods Multidrug resistance Mutation Mycobacterium tuberculosis Mycobacterium tuberculosis - drug effects Mycobacterium tuberculosis - genetics Phenotypes Pyrazinamide Pyrazinamide - pharmacology Rifampin Serum Albumin, Bovine Tuberculosis Whole Genome Sequencing |
| title | The 7H11 Agar Medium Supplemented with Calf Bovine Serum for Susceptibility Testing of Mycobacterium tuberculosis Isolates Against Pyrazinamide |
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