Supplementation of sputum cultures with culture filtrate to detect tuberculosis in a cross-sectional study of HIV-infected individuals
While some healthcare systems have shifted to molecular diagnostics, culture still remains the gold standard for tuberculosis diagnosis, but it is limited by its long duration to a positive result. Methods to reduce time to culture positivity (TTP) are urgently required. We determined if growth fact...
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Veröffentlicht in: | Tuberculosis (Edinburgh, Scotland) Scotland), 2021-07, Vol.129, p.102103-102103, Article 102103 |
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description | While some healthcare systems have shifted to molecular diagnostics, culture still remains the gold standard for tuberculosis diagnosis, but it is limited by its long duration to a positive result. Methods to reduce time to culture positivity (TTP) are urgently required. We determined if growth factor supplementation in the mycobacterial growth indicator tube (MGIT) culture system reduces TTP. MGITs were supplemented with fresh culture filtrate (CF) as a source of growth stimulatory molecules from axenic Mycobacterium tuberculosis culture. Different volumes of CF and media components were tested. The performance of these modified MGITs was assessed with sputum from HIV-TB co-infected individuals. Reducing the volume of MGIT cultures and removal of detergent from cultures grown to generate CF had a marginal but significant benefit on reducing TTP. In a subset of specimens, CF inhibited growth. Following optimization of methods, a reduced TTP occurred in specimens with low bacillary load as measured by GeneXpert, smear microscopy and colony forming units. Three specimens that were negative under standard conditions flagged positive following CF supplementation. Our data provide preliminary evidence that addition of CF to MGIT cultures can enhance detection of M. tuberculosis in HIV-TB co-infected patients with low sputum bacillary loads.
•Culture filtrate supplementation of MGIT cultures reduces time to positivity in sputum specimens with low bacterial loads.•Culture filtrate supplementation of MGITs facilitated detection in specimens flagging negative with the standard format.•A reduction in MGIT volume may increase detection but this benefit can be offset by growth inhibitory factors.•Any reduction in MGIT time to positivity was not associated with differentially culturable tubercle bacteria. |
doi_str_mv | 10.1016/j.tube.2021.102103 |
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•Culture filtrate supplementation of MGIT cultures reduces time to positivity in sputum specimens with low bacterial loads.•Culture filtrate supplementation of MGITs facilitated detection in specimens flagging negative with the standard format.•A reduction in MGIT volume may increase detection but this benefit can be offset by growth inhibitory factors.•Any reduction in MGIT time to positivity was not associated with differentially culturable tubercle bacteria.</description><identifier>ISSN: 1472-9792</identifier><identifier>EISSN: 1873-281X</identifier><identifier>DOI: 10.1016/j.tube.2021.102103</identifier><identifier>PMID: 34144375</identifier><language>eng</language><publisher>Scotland: Elsevier Ltd</publisher><subject>Adult ; Bacteriological Techniques - methods ; Coinfection ; Cross-Sectional Studies ; Culture filtrate ; Culture Media, Conditioned ; Differentially culturable tubercle bacteria ; Female ; Filtrate ; Germfree ; Growth factors ; HIV ; HIV Infections ; Human immunodeficiency virus ; Humans ; Male ; Middle Aged ; Mycobacterium tuberculosis, MGIT time To positivity ; Optimization ; Public health ; Resuscitation promoting factors ; South Africa ; Sputum ; Sputum - microbiology ; Supplements ; Tuberculosis ; Tuberculosis - diagnosis</subject><ispartof>Tuberculosis (Edinburgh, Scotland), 2021-07, Vol.129, p.102103-102103, Article 102103</ispartof><rights>2021 The Authors</rights><rights>Copyright © 2021 The Authors. Published by Elsevier Ltd.. All rights reserved.</rights><rights>Copyright Elsevier Science Ltd. Jul 2021</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c428t-5af78ee855c95020d09be8f5aa83df03b7be4b2a373be4eb04ea5836adb591093</citedby><cites>FETCH-LOGICAL-c428t-5af78ee855c95020d09be8f5aa83df03b7be4b2a373be4eb04ea5836adb591093</cites><orcidid>0000-0001-9713-3480 ; 0000-0002-7433-4383 ; 0000-0002-2940-5234 ; 0000-0001-8717-9039</orcidid></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktohtml>$$Uhttps://dx.doi.org/10.1016/j.tube.2021.102103$$EHTML$$P50$$Gelsevier$$Hfree_for_read</linktohtml><link.rule.ids>314,780,784,3548,27922,27923,45993</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/34144375$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>McIvor, Amanda</creatorcontrib><creatorcontrib>Gordhan, Bhavna Gowan</creatorcontrib><creatorcontrib>Waja, Ziyaad</creatorcontrib><creatorcontrib>Otwombe, Kennedy</creatorcontrib><creatorcontrib>Martinson, Neil A.</creatorcontrib><creatorcontrib>Kana, Bavesh Davandra</creatorcontrib><title>Supplementation of sputum cultures with culture filtrate to detect tuberculosis in a cross-sectional study of HIV-infected individuals</title><title>Tuberculosis (Edinburgh, Scotland)</title><addtitle>Tuberculosis (Edinb)</addtitle><description>While some healthcare systems have shifted to molecular diagnostics, culture still remains the gold standard for tuberculosis diagnosis, but it is limited by its long duration to a positive result. Methods to reduce time to culture positivity (TTP) are urgently required. We determined if growth factor supplementation in the mycobacterial growth indicator tube (MGIT) culture system reduces TTP. MGITs were supplemented with fresh culture filtrate (CF) as a source of growth stimulatory molecules from axenic Mycobacterium tuberculosis culture. Different volumes of CF and media components were tested. The performance of these modified MGITs was assessed with sputum from HIV-TB co-infected individuals. Reducing the volume of MGIT cultures and removal of detergent from cultures grown to generate CF had a marginal but significant benefit on reducing TTP. In a subset of specimens, CF inhibited growth. Following optimization of methods, a reduced TTP occurred in specimens with low bacillary load as measured by GeneXpert, smear microscopy and colony forming units. Three specimens that were negative under standard conditions flagged positive following CF supplementation. Our data provide preliminary evidence that addition of CF to MGIT cultures can enhance detection of M. tuberculosis in HIV-TB co-infected patients with low sputum bacillary loads.
•Culture filtrate supplementation of MGIT cultures reduces time to positivity in sputum specimens with low bacterial loads.•Culture filtrate supplementation of MGITs facilitated detection in specimens flagging negative with the standard format.•A reduction in MGIT volume may increase detection but this benefit can be offset by growth inhibitory factors.•Any reduction in MGIT time to positivity was not associated with differentially culturable tubercle bacteria.</description><subject>Adult</subject><subject>Bacteriological Techniques - methods</subject><subject>Coinfection</subject><subject>Cross-Sectional Studies</subject><subject>Culture filtrate</subject><subject>Culture Media, Conditioned</subject><subject>Differentially culturable tubercle bacteria</subject><subject>Female</subject><subject>Filtrate</subject><subject>Germfree</subject><subject>Growth factors</subject><subject>HIV</subject><subject>HIV Infections</subject><subject>Human immunodeficiency virus</subject><subject>Humans</subject><subject>Male</subject><subject>Middle Aged</subject><subject>Mycobacterium tuberculosis, MGIT time To positivity</subject><subject>Optimization</subject><subject>Public health</subject><subject>Resuscitation promoting factors</subject><subject>South Africa</subject><subject>Sputum</subject><subject>Sputum - microbiology</subject><subject>Supplements</subject><subject>Tuberculosis</subject><subject>Tuberculosis - diagnosis</subject><issn>1472-9792</issn><issn>1873-281X</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2021</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNp9kctu1TAQhi1URC_wAiwqS910k4Mv8YkjsUFHhVaqxIKL2FlOPFF9lMTBl6K-AM_NpKdlwYKVx55vfnn-n5C3nG0449t3-00uHWwEExwfBGfyBTnhupGV0PzHEdZ1I6q2acUxOU1pz3CIafaKHMua17Vs1An5_aUsywgTzNlmH2YaBpqWkstE-zLmEiHRXz7fPd_o4MccbQaaA3WQoc90_UXEfkg-UT9TS_sYUqoSNlHSjjTl4h5W6eub75WfB2yAQ9T5e--KHdNr8nLAA948nWfk28err7vr6vbzp5vdh9uqr4XOlbJDowG0Un2rmGCOtR3oQVmrpRuY7JoO6k5Y2UgsoGM1WKXl1rpOtZy18oxcHnSXGH4WSNlMPvUwjnaGUJIRqpbojGgbRC_-QfehRNxmpbaaSaa3KyUO1OPKEQazRD_Z-GA4M2tKZm9Wf8yakjmkhEPnT9Klm8D9HXmOBYH3BwDQi3sP0aTew9yD8xG9My74_-n_Ae7WpiE</recordid><startdate>202107</startdate><enddate>202107</enddate><creator>McIvor, Amanda</creator><creator>Gordhan, Bhavna Gowan</creator><creator>Waja, Ziyaad</creator><creator>Otwombe, Kennedy</creator><creator>Martinson, Neil A.</creator><creator>Kana, Bavesh Davandra</creator><general>Elsevier Ltd</general><general>Elsevier Science Ltd</general><scope>6I.</scope><scope>AAFTH</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>C1K</scope><scope>7X8</scope><orcidid>https://orcid.org/0000-0001-9713-3480</orcidid><orcidid>https://orcid.org/0000-0002-7433-4383</orcidid><orcidid>https://orcid.org/0000-0002-2940-5234</orcidid><orcidid>https://orcid.org/0000-0001-8717-9039</orcidid></search><sort><creationdate>202107</creationdate><title>Supplementation of sputum cultures with culture filtrate to detect tuberculosis in a cross-sectional study of HIV-infected individuals</title><author>McIvor, Amanda ; Gordhan, Bhavna Gowan ; Waja, Ziyaad ; Otwombe, Kennedy ; Martinson, Neil A. ; Kana, Bavesh Davandra</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c428t-5af78ee855c95020d09be8f5aa83df03b7be4b2a373be4eb04ea5836adb591093</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2021</creationdate><topic>Adult</topic><topic>Bacteriological Techniques - methods</topic><topic>Coinfection</topic><topic>Cross-Sectional Studies</topic><topic>Culture filtrate</topic><topic>Culture Media, Conditioned</topic><topic>Differentially culturable tubercle bacteria</topic><topic>Female</topic><topic>Filtrate</topic><topic>Germfree</topic><topic>Growth factors</topic><topic>HIV</topic><topic>HIV Infections</topic><topic>Human immunodeficiency virus</topic><topic>Humans</topic><topic>Male</topic><topic>Middle Aged</topic><topic>Mycobacterium tuberculosis, MGIT time To positivity</topic><topic>Optimization</topic><topic>Public health</topic><topic>Resuscitation promoting factors</topic><topic>South Africa</topic><topic>Sputum</topic><topic>Sputum - microbiology</topic><topic>Supplements</topic><topic>Tuberculosis</topic><topic>Tuberculosis - diagnosis</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>McIvor, Amanda</creatorcontrib><creatorcontrib>Gordhan, Bhavna Gowan</creatorcontrib><creatorcontrib>Waja, Ziyaad</creatorcontrib><creatorcontrib>Otwombe, Kennedy</creatorcontrib><creatorcontrib>Martinson, Neil A.</creatorcontrib><creatorcontrib>Kana, Bavesh Davandra</creatorcontrib><collection>ScienceDirect Open Access Titles</collection><collection>Elsevier:ScienceDirect:Open Access</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>Environmental Sciences and Pollution Management</collection><collection>MEDLINE - Academic</collection><jtitle>Tuberculosis (Edinburgh, Scotland)</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>McIvor, Amanda</au><au>Gordhan, Bhavna Gowan</au><au>Waja, Ziyaad</au><au>Otwombe, Kennedy</au><au>Martinson, Neil A.</au><au>Kana, Bavesh Davandra</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Supplementation of sputum cultures with culture filtrate to detect tuberculosis in a cross-sectional study of HIV-infected individuals</atitle><jtitle>Tuberculosis (Edinburgh, Scotland)</jtitle><addtitle>Tuberculosis (Edinb)</addtitle><date>2021-07</date><risdate>2021</risdate><volume>129</volume><spage>102103</spage><epage>102103</epage><pages>102103-102103</pages><artnum>102103</artnum><issn>1472-9792</issn><eissn>1873-281X</eissn><abstract>While some healthcare systems have shifted to molecular diagnostics, culture still remains the gold standard for tuberculosis diagnosis, but it is limited by its long duration to a positive result. Methods to reduce time to culture positivity (TTP) are urgently required. We determined if growth factor supplementation in the mycobacterial growth indicator tube (MGIT) culture system reduces TTP. MGITs were supplemented with fresh culture filtrate (CF) as a source of growth stimulatory molecules from axenic Mycobacterium tuberculosis culture. Different volumes of CF and media components were tested. The performance of these modified MGITs was assessed with sputum from HIV-TB co-infected individuals. Reducing the volume of MGIT cultures and removal of detergent from cultures grown to generate CF had a marginal but significant benefit on reducing TTP. In a subset of specimens, CF inhibited growth. Following optimization of methods, a reduced TTP occurred in specimens with low bacillary load as measured by GeneXpert, smear microscopy and colony forming units. Three specimens that were negative under standard conditions flagged positive following CF supplementation. Our data provide preliminary evidence that addition of CF to MGIT cultures can enhance detection of M. tuberculosis in HIV-TB co-infected patients with low sputum bacillary loads.
•Culture filtrate supplementation of MGIT cultures reduces time to positivity in sputum specimens with low bacterial loads.•Culture filtrate supplementation of MGITs facilitated detection in specimens flagging negative with the standard format.•A reduction in MGIT volume may increase detection but this benefit can be offset by growth inhibitory factors.•Any reduction in MGIT time to positivity was not associated with differentially culturable tubercle bacteria.</abstract><cop>Scotland</cop><pub>Elsevier Ltd</pub><pmid>34144375</pmid><doi>10.1016/j.tube.2021.102103</doi><tpages>1</tpages><orcidid>https://orcid.org/0000-0001-9713-3480</orcidid><orcidid>https://orcid.org/0000-0002-7433-4383</orcidid><orcidid>https://orcid.org/0000-0002-2940-5234</orcidid><orcidid>https://orcid.org/0000-0001-8717-9039</orcidid><oa>free_for_read</oa></addata></record> |
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subjects | Adult Bacteriological Techniques - methods Coinfection Cross-Sectional Studies Culture filtrate Culture Media, Conditioned Differentially culturable tubercle bacteria Female Filtrate Germfree Growth factors HIV HIV Infections Human immunodeficiency virus Humans Male Middle Aged Mycobacterium tuberculosis, MGIT time To positivity Optimization Public health Resuscitation promoting factors South Africa Sputum Sputum - microbiology Supplements Tuberculosis Tuberculosis - diagnosis |
title | Supplementation of sputum cultures with culture filtrate to detect tuberculosis in a cross-sectional study of HIV-infected individuals |
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