Integrated nucleic acid testing system to enable TB diagnosis in peripheral settings
To facilitate treatment and limit transmission of tuberculosis (TB), new methods are needed to enable rapid and affordable diagnosis of the disease in high-burden low-resource settings. We have developed a prototype integrated nucleic acid testing device to detect Mycobacterium tuberculosis ( M.tb )...
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| Veröffentlicht in: | Lab on a chip 2020-10, Vol.2 (21), p.471-481 |
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| creator | Lu, Hsiang-Wei Sakamuri, Rama Kumar, Pranav Ferguson, Tanya M Doebler, Robert W Herrington, Keith D Talbot, Ryan P Weigel, Kris M Nguyen, Felicia K Cangelosi, Gerard A Narita, Masahiro Boyle, David S Niemz, Angelika |
| description | To facilitate treatment and limit transmission of tuberculosis (TB), new methods are needed to enable rapid and affordable diagnosis of the disease in high-burden low-resource settings. We have developed a prototype integrated nucleic acid testing device to detect
Mycobacterium tuberculosis
(
M.tb
) in sputum. The device consists of a disposable cartridge and compact, inexpensive instrument that automates pathogen lysis, nucleic acid extraction, isothermal DNA amplification and lateral flow detection. A liquefied and disinfected sputum sample is manually injected into the cartridge, and all other steps are automated, with a result provided in |
| doi_str_mv | 10.1039/d0lc00445f |
| format | Article |
| fullrecord | <record><control><sourceid>proquest_pubme</sourceid><recordid>TN_cdi_pubmedcentral_primary_oai_pubmedcentral_nih_gov_7787164</recordid><sourceformat>XML</sourceformat><sourcesystem>PC</sourcesystem><sourcerecordid>2454400530</sourcerecordid><originalsourceid>FETCH-LOGICAL-c532t-535299aed09d1d11c88d80dae17b372951e03c674087f3ae9174b458705831013</originalsourceid><addsrcrecordid>eNp9kc1vEzEQxS1ERT_gwh1k1BtSyszajr2XShAorRSJSzhbjj2butp4F9tB6n_PtikBLpw80vvN87MfY68RLhBE-yFA7wGkVN0zdoJSixmgaZ8f5lYfs9NS7gBQybl5wY6FgAbniCdsdZMqbbKrFHja-Z6i587HwCuVGtOGl_tSacvrwCm5dU989YmH6DZpKLHwmPhIOY63lF3PC9WHnfKSHXWuL_Tq6Txj36--rBbXs-W3rzeLj8uZV6KpMyVU07aOArQBA6I3JhgIjlCvhW5ahQTCz7UEozvhqEUt11IZDcoIBBRn7HLvO-7WWwqeUp1i2DHHrcv3dnDR_qukeGs3w0-rtdE4l5PB-ZNBHn7sphfbu2GX05TZNlJJCaAETNT7PeXzUEqm7nADgn1owH6G5eKxgasJfvt3pgP6-8sn4M0eyMUf1D8VTvq7_-l2DJ34BYm1lgs</addsrcrecordid><sourcetype>Open Access Repository</sourcetype><iscdi>true</iscdi><recordtype>article</recordtype><pqid>2454400530</pqid></control><display><type>article</type><title>Integrated nucleic acid testing system to enable TB diagnosis in peripheral settings</title><source>Royal Society Of Chemistry Journals 2008-</source><source>Alma/SFX Local Collection</source><creator>Lu, Hsiang-Wei ; Sakamuri, Rama ; Kumar, Pranav ; Ferguson, Tanya M ; Doebler, Robert W ; Herrington, Keith D ; Talbot, Ryan P ; Weigel, Kris M ; Nguyen, Felicia K ; Cangelosi, Gerard A ; Narita, Masahiro ; Boyle, David S ; Niemz, Angelika</creator><creatorcontrib>Lu, Hsiang-Wei ; Sakamuri, Rama ; Kumar, Pranav ; Ferguson, Tanya M ; Doebler, Robert W ; Herrington, Keith D ; Talbot, Ryan P ; Weigel, Kris M ; Nguyen, Felicia K ; Cangelosi, Gerard A ; Narita, Masahiro ; Boyle, David S ; Niemz, Angelika</creatorcontrib><description>To facilitate treatment and limit transmission of tuberculosis (TB), new methods are needed to enable rapid and affordable diagnosis of the disease in high-burden low-resource settings. We have developed a prototype integrated nucleic acid testing device to detect
Mycobacterium tuberculosis
(
M.tb
) in sputum. The device consists of a disposable cartridge and compact, inexpensive instrument that automates pathogen lysis, nucleic acid extraction, isothermal DNA amplification and lateral flow detection. A liquefied and disinfected sputum sample is manually injected into the cartridge, and all other steps are automated, with a result provided in <1.5 h. Cell disruption and DNA extraction is executed within a four-port active valve containing a miniature bead blender (based on PureLyse® technology, Claremont BioSolutions LLC). The DNA-containing eluate is combined with dry master-mix reagents and target DNA is isothermally amplified. Amplified master-mix is then pumped into a lateral flow strip chamber for detection. The entire process is performed in a single-use closed-system cartridge to prevent amplicon carryover. For testing of
M.tb
-spiked sputum the system provided a limit of detection of 5 × 10
3
colony forming units (CFU) per mL. None of the negative sputum-only controls yielded a false-positive result. Testing of 45 clinical sputum specimens from TB cases and controls relative to a validated manual qPCR-based comparator method revealed a preliminary sensitivity of 90% and specificity of 96%. With further development, the herein described integrated nucleic acid testing device can enable TB diagnosis and treatment initiation in the same clinical encounter in near-patient low-resource settings of high TB burden countries.
We developed a nucleic acid testing device that automates pathogen lysis, DNA extraction, isothermal DNA amplification and lateral flow detection.</description><identifier>ISSN: 1473-0197</identifier><identifier>EISSN: 1473-0189</identifier><identifier>DOI: 10.1039/d0lc00445f</identifier><identifier>PMID: 33021611</identifier><language>eng</language><publisher>England: Royal Society of Chemistry</publisher><subject>Amplification ; Automation ; Cartridges ; Deoxyribonucleic acid ; Diagnosis ; DNA ; Health services ; Reagents ; Tuberculosis</subject><ispartof>Lab on a chip, 2020-10, Vol.2 (21), p.471-481</ispartof><rights>Copyright Royal Society of Chemistry 2020</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c532t-535299aed09d1d11c88d80dae17b372951e03c674087f3ae9174b458705831013</citedby><cites>FETCH-LOGICAL-c532t-535299aed09d1d11c88d80dae17b372951e03c674087f3ae9174b458705831013</cites><orcidid>0000-0001-7385-1541 ; 0000-0002-9135-2217</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/33021611$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Lu, Hsiang-Wei</creatorcontrib><creatorcontrib>Sakamuri, Rama</creatorcontrib><creatorcontrib>Kumar, Pranav</creatorcontrib><creatorcontrib>Ferguson, Tanya M</creatorcontrib><creatorcontrib>Doebler, Robert W</creatorcontrib><creatorcontrib>Herrington, Keith D</creatorcontrib><creatorcontrib>Talbot, Ryan P</creatorcontrib><creatorcontrib>Weigel, Kris M</creatorcontrib><creatorcontrib>Nguyen, Felicia K</creatorcontrib><creatorcontrib>Cangelosi, Gerard A</creatorcontrib><creatorcontrib>Narita, Masahiro</creatorcontrib><creatorcontrib>Boyle, David S</creatorcontrib><creatorcontrib>Niemz, Angelika</creatorcontrib><title>Integrated nucleic acid testing system to enable TB diagnosis in peripheral settings</title><title>Lab on a chip</title><addtitle>Lab Chip</addtitle><description>To facilitate treatment and limit transmission of tuberculosis (TB), new methods are needed to enable rapid and affordable diagnosis of the disease in high-burden low-resource settings. We have developed a prototype integrated nucleic acid testing device to detect
Mycobacterium tuberculosis
(
M.tb
) in sputum. The device consists of a disposable cartridge and compact, inexpensive instrument that automates pathogen lysis, nucleic acid extraction, isothermal DNA amplification and lateral flow detection. A liquefied and disinfected sputum sample is manually injected into the cartridge, and all other steps are automated, with a result provided in <1.5 h. Cell disruption and DNA extraction is executed within a four-port active valve containing a miniature bead blender (based on PureLyse® technology, Claremont BioSolutions LLC). The DNA-containing eluate is combined with dry master-mix reagents and target DNA is isothermally amplified. Amplified master-mix is then pumped into a lateral flow strip chamber for detection. The entire process is performed in a single-use closed-system cartridge to prevent amplicon carryover. For testing of
M.tb
-spiked sputum the system provided a limit of detection of 5 × 10
3
colony forming units (CFU) per mL. None of the negative sputum-only controls yielded a false-positive result. Testing of 45 clinical sputum specimens from TB cases and controls relative to a validated manual qPCR-based comparator method revealed a preliminary sensitivity of 90% and specificity of 96%. With further development, the herein described integrated nucleic acid testing device can enable TB diagnosis and treatment initiation in the same clinical encounter in near-patient low-resource settings of high TB burden countries.
We developed a nucleic acid testing device that automates pathogen lysis, DNA extraction, isothermal DNA amplification and lateral flow detection.</description><subject>Amplification</subject><subject>Automation</subject><subject>Cartridges</subject><subject>Deoxyribonucleic acid</subject><subject>Diagnosis</subject><subject>DNA</subject><subject>Health services</subject><subject>Reagents</subject><subject>Tuberculosis</subject><issn>1473-0197</issn><issn>1473-0189</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2020</creationdate><recordtype>article</recordtype><recordid>eNp9kc1vEzEQxS1ERT_gwh1k1BtSyszajr2XShAorRSJSzhbjj2butp4F9tB6n_PtikBLpw80vvN87MfY68RLhBE-yFA7wGkVN0zdoJSixmgaZ8f5lYfs9NS7gBQybl5wY6FgAbniCdsdZMqbbKrFHja-Z6i587HwCuVGtOGl_tSacvrwCm5dU989YmH6DZpKLHwmPhIOY63lF3PC9WHnfKSHXWuL_Tq6Txj36--rBbXs-W3rzeLj8uZV6KpMyVU07aOArQBA6I3JhgIjlCvhW5ahQTCz7UEozvhqEUt11IZDcoIBBRn7HLvO-7WWwqeUp1i2DHHrcv3dnDR_qukeGs3w0-rtdE4l5PB-ZNBHn7sphfbu2GX05TZNlJJCaAETNT7PeXzUEqm7nADgn1owH6G5eKxgasJfvt3pgP6-8sn4M0eyMUf1D8VTvq7_-l2DJ34BYm1lgs</recordid><startdate>20201027</startdate><enddate>20201027</enddate><creator>Lu, Hsiang-Wei</creator><creator>Sakamuri, Rama</creator><creator>Kumar, Pranav</creator><creator>Ferguson, Tanya M</creator><creator>Doebler, Robert W</creator><creator>Herrington, Keith D</creator><creator>Talbot, Ryan P</creator><creator>Weigel, Kris M</creator><creator>Nguyen, Felicia K</creator><creator>Cangelosi, Gerard A</creator><creator>Narita, Masahiro</creator><creator>Boyle, David S</creator><creator>Niemz, Angelika</creator><general>Royal Society of Chemistry</general><scope>NPM</scope><scope>AAYXX</scope><scope>CITATION</scope><scope>7SP</scope><scope>7TB</scope><scope>7U5</scope><scope>8FD</scope><scope>FR3</scope><scope>L7M</scope><scope>5PM</scope><orcidid>https://orcid.org/0000-0001-7385-1541</orcidid><orcidid>https://orcid.org/0000-0002-9135-2217</orcidid></search><sort><creationdate>20201027</creationdate><title>Integrated nucleic acid testing system to enable TB diagnosis in peripheral settings</title><author>Lu, Hsiang-Wei ; Sakamuri, Rama ; Kumar, Pranav ; Ferguson, Tanya M ; Doebler, Robert W ; Herrington, Keith D ; Talbot, Ryan P ; Weigel, Kris M ; Nguyen, Felicia K ; Cangelosi, Gerard A ; Narita, Masahiro ; Boyle, David S ; Niemz, Angelika</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c532t-535299aed09d1d11c88d80dae17b372951e03c674087f3ae9174b458705831013</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2020</creationdate><topic>Amplification</topic><topic>Automation</topic><topic>Cartridges</topic><topic>Deoxyribonucleic acid</topic><topic>Diagnosis</topic><topic>DNA</topic><topic>Health services</topic><topic>Reagents</topic><topic>Tuberculosis</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Lu, Hsiang-Wei</creatorcontrib><creatorcontrib>Sakamuri, Rama</creatorcontrib><creatorcontrib>Kumar, Pranav</creatorcontrib><creatorcontrib>Ferguson, Tanya M</creatorcontrib><creatorcontrib>Doebler, Robert W</creatorcontrib><creatorcontrib>Herrington, Keith D</creatorcontrib><creatorcontrib>Talbot, Ryan P</creatorcontrib><creatorcontrib>Weigel, Kris M</creatorcontrib><creatorcontrib>Nguyen, Felicia K</creatorcontrib><creatorcontrib>Cangelosi, Gerard A</creatorcontrib><creatorcontrib>Narita, Masahiro</creatorcontrib><creatorcontrib>Boyle, David S</creatorcontrib><creatorcontrib>Niemz, Angelika</creatorcontrib><collection>PubMed</collection><collection>CrossRef</collection><collection>Electronics & Communications Abstracts</collection><collection>Mechanical & Transportation Engineering Abstracts</collection><collection>Solid State and Superconductivity Abstracts</collection><collection>Technology Research Database</collection><collection>Engineering Research Database</collection><collection>Advanced Technologies Database with Aerospace</collection><collection>PubMed Central (Full Participant titles)</collection><jtitle>Lab on a chip</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Lu, Hsiang-Wei</au><au>Sakamuri, Rama</au><au>Kumar, Pranav</au><au>Ferguson, Tanya M</au><au>Doebler, Robert W</au><au>Herrington, Keith D</au><au>Talbot, Ryan P</au><au>Weigel, Kris M</au><au>Nguyen, Felicia K</au><au>Cangelosi, Gerard A</au><au>Narita, Masahiro</au><au>Boyle, David S</au><au>Niemz, Angelika</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Integrated nucleic acid testing system to enable TB diagnosis in peripheral settings</atitle><jtitle>Lab on a chip</jtitle><addtitle>Lab Chip</addtitle><date>2020-10-27</date><risdate>2020</risdate><volume>2</volume><issue>21</issue><spage>471</spage><epage>481</epage><pages>471-481</pages><issn>1473-0197</issn><eissn>1473-0189</eissn><abstract>To facilitate treatment and limit transmission of tuberculosis (TB), new methods are needed to enable rapid and affordable diagnosis of the disease in high-burden low-resource settings. We have developed a prototype integrated nucleic acid testing device to detect
Mycobacterium tuberculosis
(
M.tb
) in sputum. The device consists of a disposable cartridge and compact, inexpensive instrument that automates pathogen lysis, nucleic acid extraction, isothermal DNA amplification and lateral flow detection. A liquefied and disinfected sputum sample is manually injected into the cartridge, and all other steps are automated, with a result provided in <1.5 h. Cell disruption and DNA extraction is executed within a four-port active valve containing a miniature bead blender (based on PureLyse® technology, Claremont BioSolutions LLC). The DNA-containing eluate is combined with dry master-mix reagents and target DNA is isothermally amplified. Amplified master-mix is then pumped into a lateral flow strip chamber for detection. The entire process is performed in a single-use closed-system cartridge to prevent amplicon carryover. For testing of
M.tb
-spiked sputum the system provided a limit of detection of 5 × 10
3
colony forming units (CFU) per mL. None of the negative sputum-only controls yielded a false-positive result. Testing of 45 clinical sputum specimens from TB cases and controls relative to a validated manual qPCR-based comparator method revealed a preliminary sensitivity of 90% and specificity of 96%. With further development, the herein described integrated nucleic acid testing device can enable TB diagnosis and treatment initiation in the same clinical encounter in near-patient low-resource settings of high TB burden countries.
We developed a nucleic acid testing device that automates pathogen lysis, DNA extraction, isothermal DNA amplification and lateral flow detection.</abstract><cop>England</cop><pub>Royal Society of Chemistry</pub><pmid>33021611</pmid><doi>10.1039/d0lc00445f</doi><tpages>11</tpages><orcidid>https://orcid.org/0000-0001-7385-1541</orcidid><orcidid>https://orcid.org/0000-0002-9135-2217</orcidid><oa>free_for_read</oa></addata></record> |
| fulltext | fulltext |
| identifier | ISSN: 1473-0197 |
| ispartof | Lab on a chip, 2020-10, Vol.2 (21), p.471-481 |
| issn | 1473-0197 1473-0189 |
| language | eng |
| recordid | cdi_pubmedcentral_primary_oai_pubmedcentral_nih_gov_7787164 |
| source | Royal Society Of Chemistry Journals 2008-; Alma/SFX Local Collection |
| subjects | Amplification Automation Cartridges Deoxyribonucleic acid Diagnosis DNA Health services Reagents Tuberculosis |
| title | Integrated nucleic acid testing system to enable TB diagnosis in peripheral settings |
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