Enhancing Loop-Mediated Isothermal Amplification through Nonpowered Nanoelectric Preconcentration
The global threat posed by the COVID-19 pandemic has catalyzed the development of point-of-care (POC) molecular diagnostics. While loop-mediated isothermal amplification (LAMP) stands out as a promising technique among FDA-approved methods, it is occasionally susceptible to a high risk of false posi...
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| Veröffentlicht in: | Analytical chemistry (Washington) 2024-03, Vol.96 (9), p.3844-3852 |
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| creator | Lee, Na Eun Kim, Kang Hyeon Cho, Youngkyu Kim, Jinhwan Kwak, Seungmin Lee, Dohwan Yoon, Dae Sung Lee, Jeong Hoon |
| description | The global threat posed by the COVID-19 pandemic has catalyzed the development of point-of-care (POC) molecular diagnostics. While loop-mediated isothermal amplification (LAMP) stands out as a promising technique among FDA-approved methods, it is occasionally susceptible to a high risk of false positives due to nonspecific amplification of a primer dimer. In this work, we report an enhancing LAMP technique in terms of assay sensitivity and reliability through streamlined integration with a nonpowered nanoelectric preconcentration (NPP). The NPP, serving as a sample preparation tool, enriched the virus concentration in samples prior to the subsequent LAMP assay. This enrichment enabled not only to achieve more sensitive assay but also to shorten the assay time for all tested clinical samples by ∼10 min compared to the conventional LAMP. The shortened assay time suppresses the occurrence of nonspecific amplification by not providing the necessary incubation time, effectively suppressing misidentification by false positives. Utilizing this technique, we also developed a prototype of the POC NPP-LAMP kit. This kit offers a streamlined diagnostic process for nontrained individuals, from the sample enrichment, transfer of the enriched sample to LAMP assays, which facilitates on-site/on-demand diagnosis of SARS-CoV-2. This development holds the potential to contribute toward preventing not only the current outbreak but also future occurrences of pandemic viruses. |
| doi_str_mv | 10.1021/acs.analchem.3c05236 |
| format | Article |
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While loop-mediated isothermal amplification (LAMP) stands out as a promising technique among FDA-approved methods, it is occasionally susceptible to a high risk of false positives due to nonspecific amplification of a primer dimer. In this work, we report an enhancing LAMP technique in terms of assay sensitivity and reliability through streamlined integration with a nonpowered nanoelectric preconcentration (NPP). The NPP, serving as a sample preparation tool, enriched the virus concentration in samples prior to the subsequent LAMP assay. This enrichment enabled not only to achieve more sensitive assay but also to shorten the assay time for all tested clinical samples by ∼10 min compared to the conventional LAMP. The shortened assay time suppresses the occurrence of nonspecific amplification by not providing the necessary incubation time, effectively suppressing misidentification by false positives. Utilizing this technique, we also developed a prototype of the POC NPP-LAMP kit. This kit offers a streamlined diagnostic process for nontrained individuals, from the sample enrichment, transfer of the enriched sample to LAMP assays, which facilitates on-site/on-demand diagnosis of SARS-CoV-2. This development holds the potential to contribute toward preventing not only the current outbreak but also future occurrences of pandemic viruses.</description><identifier>ISSN: 0003-2700</identifier><identifier>ISSN: 1520-6882</identifier><identifier>EISSN: 1520-6882</identifier><identifier>DOI: 10.1021/acs.analchem.3c05236</identifier><identifier>PMID: 38393745</identifier><language>eng</language><publisher>United States: American Chemical Society</publisher><subject>analytical chemistry ; Assaying ; COVID-19 ; COVID-19 - diagnosis ; COVID-19 infection ; Enrichment ; Gene amplification ; Humans ; loop-mediated isothermal amplification ; Molecular Diagnostic Techniques - methods ; Nucleic Acid Amplification Techniques - methods ; pandemic ; Pandemics ; point-of-care systems ; prototypes ; Reproducibility of Results ; risk ; RNA, Viral ; Sample preparation ; Sensitivity and Specificity ; Severe acute respiratory syndrome coronavirus 2 ; Viral diseases ; viruses</subject><ispartof>Analytical chemistry (Washington), 2024-03, Vol.96 (9), p.3844-3852</ispartof><rights>2024 American Chemical Society</rights><rights>Copyright American Chemical Society Mar 5, 2024</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><cites>FETCH-LOGICAL-a358t-10b30aec48a74985b2a94e8cd77a15a98d0fe9b29f325947580981e0f8b630593</cites><orcidid>0000-0001-6002-1557</orcidid></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://pubs.acs.org/doi/pdf/10.1021/acs.analchem.3c05236$$EPDF$$P50$$Gacs$$H</linktopdf><linktohtml>$$Uhttps://pubs.acs.org/doi/10.1021/acs.analchem.3c05236$$EHTML$$P50$$Gacs$$H</linktohtml><link.rule.ids>314,776,780,2752,27053,27901,27902,56713,56763</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/38393745$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Lee, Na Eun</creatorcontrib><creatorcontrib>Kim, Kang Hyeon</creatorcontrib><creatorcontrib>Cho, Youngkyu</creatorcontrib><creatorcontrib>Kim, Jinhwan</creatorcontrib><creatorcontrib>Kwak, Seungmin</creatorcontrib><creatorcontrib>Lee, Dohwan</creatorcontrib><creatorcontrib>Yoon, Dae Sung</creatorcontrib><creatorcontrib>Lee, Jeong Hoon</creatorcontrib><title>Enhancing Loop-Mediated Isothermal Amplification through Nonpowered Nanoelectric Preconcentration</title><title>Analytical chemistry (Washington)</title><addtitle>Anal. Chem</addtitle><description>The global threat posed by the COVID-19 pandemic has catalyzed the development of point-of-care (POC) molecular diagnostics. While loop-mediated isothermal amplification (LAMP) stands out as a promising technique among FDA-approved methods, it is occasionally susceptible to a high risk of false positives due to nonspecific amplification of a primer dimer. In this work, we report an enhancing LAMP technique in terms of assay sensitivity and reliability through streamlined integration with a nonpowered nanoelectric preconcentration (NPP). The NPP, serving as a sample preparation tool, enriched the virus concentration in samples prior to the subsequent LAMP assay. This enrichment enabled not only to achieve more sensitive assay but also to shorten the assay time for all tested clinical samples by ∼10 min compared to the conventional LAMP. The shortened assay time suppresses the occurrence of nonspecific amplification by not providing the necessary incubation time, effectively suppressing misidentification by false positives. Utilizing this technique, we also developed a prototype of the POC NPP-LAMP kit. This kit offers a streamlined diagnostic process for nontrained individuals, from the sample enrichment, transfer of the enriched sample to LAMP assays, which facilitates on-site/on-demand diagnosis of SARS-CoV-2. This development holds the potential to contribute toward preventing not only the current outbreak but also future occurrences of pandemic viruses.</description><subject>analytical chemistry</subject><subject>Assaying</subject><subject>COVID-19</subject><subject>COVID-19 - diagnosis</subject><subject>COVID-19 infection</subject><subject>Enrichment</subject><subject>Gene amplification</subject><subject>Humans</subject><subject>loop-mediated isothermal amplification</subject><subject>Molecular Diagnostic Techniques - methods</subject><subject>Nucleic Acid Amplification Techniques - methods</subject><subject>pandemic</subject><subject>Pandemics</subject><subject>point-of-care systems</subject><subject>prototypes</subject><subject>Reproducibility of Results</subject><subject>risk</subject><subject>RNA, Viral</subject><subject>Sample preparation</subject><subject>Sensitivity and Specificity</subject><subject>Severe acute respiratory syndrome coronavirus 2</subject><subject>Viral diseases</subject><subject>viruses</subject><issn>0003-2700</issn><issn>1520-6882</issn><issn>1520-6882</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2024</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNqFkU1v1DAURS1ERYfCP0AoEptuMn3-SuxlVZVSaWi7gHX04rw0qRI72IkQ_54MM-2CBaze5tz7pHsY-8Bhy0HwC3Rpix4H19G4lQ60kMUrtuFaQF4YI16zDQDIXJQAp-xtSk8AnAMv3rBTaaSVpdIbhte-Q-96_5jtQpjyr9T0OFOT3aYwdxRHHLLLcRr6tnc498FncxfD8thld8FP4SfFlb1DH2ggN8feZQ-RXPCO_Bz_BN6xkxaHRO-P94x9_3z97epLvru_ub263OUotZlzDrUEJKcMlsoaXQu0ioxryhK5RmsaaMnWwrZSaKtKbcAaTtCaupCgrTxj54feKYYfC6W5GvvkaBjQU1hSJUGBEsqC-C8q1nFKKEHoFf30F_oUlrjOvqeUVCUXolgpdaBcDClFaqsp9iPGXxWHam-rWm1Vz7aqo6019vFYvtQjNS-hZz0rAAdgH395_M_O32duo6M</recordid><startdate>20240305</startdate><enddate>20240305</enddate><creator>Lee, Na Eun</creator><creator>Kim, Kang Hyeon</creator><creator>Cho, Youngkyu</creator><creator>Kim, Jinhwan</creator><creator>Kwak, Seungmin</creator><creator>Lee, Dohwan</creator><creator>Yoon, Dae Sung</creator><creator>Lee, Jeong Hoon</creator><general>American Chemical Society</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>7QF</scope><scope>7QO</scope><scope>7QQ</scope><scope>7SC</scope><scope>7SE</scope><scope>7SP</scope><scope>7SR</scope><scope>7TA</scope><scope>7TB</scope><scope>7TM</scope><scope>7U5</scope><scope>7U7</scope><scope>7U9</scope><scope>8BQ</scope><scope>8FD</scope><scope>C1K</scope><scope>F28</scope><scope>FR3</scope><scope>H8D</scope><scope>H8G</scope><scope>H94</scope><scope>JG9</scope><scope>JQ2</scope><scope>KR7</scope><scope>L7M</scope><scope>L~C</scope><scope>L~D</scope><scope>P64</scope><scope>7X8</scope><scope>7S9</scope><scope>L.6</scope><orcidid>https://orcid.org/0000-0001-6002-1557</orcidid></search><sort><creationdate>20240305</creationdate><title>Enhancing Loop-Mediated Isothermal Amplification through Nonpowered Nanoelectric Preconcentration</title><author>Lee, Na Eun ; Kim, Kang Hyeon ; Cho, Youngkyu ; Kim, Jinhwan ; Kwak, Seungmin ; Lee, Dohwan ; Yoon, Dae Sung ; Lee, Jeong Hoon</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-a358t-10b30aec48a74985b2a94e8cd77a15a98d0fe9b29f325947580981e0f8b630593</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2024</creationdate><topic>analytical chemistry</topic><topic>Assaying</topic><topic>COVID-19</topic><topic>COVID-19 - diagnosis</topic><topic>COVID-19 infection</topic><topic>Enrichment</topic><topic>Gene amplification</topic><topic>Humans</topic><topic>loop-mediated isothermal amplification</topic><topic>Molecular Diagnostic Techniques - methods</topic><topic>Nucleic Acid Amplification Techniques - methods</topic><topic>pandemic</topic><topic>Pandemics</topic><topic>point-of-care systems</topic><topic>prototypes</topic><topic>Reproducibility of Results</topic><topic>risk</topic><topic>RNA, Viral</topic><topic>Sample preparation</topic><topic>Sensitivity and Specificity</topic><topic>Severe acute respiratory syndrome coronavirus 2</topic><topic>Viral diseases</topic><topic>viruses</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Lee, Na Eun</creatorcontrib><creatorcontrib>Kim, Kang Hyeon</creatorcontrib><creatorcontrib>Cho, Youngkyu</creatorcontrib><creatorcontrib>Kim, Jinhwan</creatorcontrib><creatorcontrib>Kwak, Seungmin</creatorcontrib><creatorcontrib>Lee, Dohwan</creatorcontrib><creatorcontrib>Yoon, Dae Sung</creatorcontrib><creatorcontrib>Lee, Jeong Hoon</creatorcontrib><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>Aluminium Industry Abstracts</collection><collection>Biotechnology Research Abstracts</collection><collection>Ceramic Abstracts</collection><collection>Computer and Information Systems Abstracts</collection><collection>Corrosion Abstracts</collection><collection>Electronics & Communications Abstracts</collection><collection>Engineered Materials Abstracts</collection><collection>Materials Business File</collection><collection>Mechanical & Transportation Engineering Abstracts</collection><collection>Nucleic Acids Abstracts</collection><collection>Solid State and Superconductivity Abstracts</collection><collection>Toxicology Abstracts</collection><collection>Virology and AIDS Abstracts</collection><collection>METADEX</collection><collection>Technology Research Database</collection><collection>Environmental Sciences and Pollution Management</collection><collection>ANTE: Abstracts in New Technology & Engineering</collection><collection>Engineering Research Database</collection><collection>Aerospace Database</collection><collection>Copper Technical Reference Library</collection><collection>AIDS and Cancer Research Abstracts</collection><collection>Materials Research Database</collection><collection>ProQuest Computer Science Collection</collection><collection>Civil Engineering Abstracts</collection><collection>Advanced Technologies Database with Aerospace</collection><collection>Computer and Information Systems Abstracts Academic</collection><collection>Computer and Information Systems Abstracts Professional</collection><collection>Biotechnology and BioEngineering Abstracts</collection><collection>MEDLINE - Academic</collection><collection>AGRICOLA</collection><collection>AGRICOLA - Academic</collection><jtitle>Analytical chemistry (Washington)</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Lee, Na Eun</au><au>Kim, Kang Hyeon</au><au>Cho, Youngkyu</au><au>Kim, Jinhwan</au><au>Kwak, Seungmin</au><au>Lee, Dohwan</au><au>Yoon, Dae Sung</au><au>Lee, Jeong Hoon</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Enhancing Loop-Mediated Isothermal Amplification through Nonpowered Nanoelectric Preconcentration</atitle><jtitle>Analytical chemistry (Washington)</jtitle><addtitle>Anal. Chem</addtitle><date>2024-03-05</date><risdate>2024</risdate><volume>96</volume><issue>9</issue><spage>3844</spage><epage>3852</epage><pages>3844-3852</pages><issn>0003-2700</issn><issn>1520-6882</issn><eissn>1520-6882</eissn><abstract>The global threat posed by the COVID-19 pandemic has catalyzed the development of point-of-care (POC) molecular diagnostics. While loop-mediated isothermal amplification (LAMP) stands out as a promising technique among FDA-approved methods, it is occasionally susceptible to a high risk of false positives due to nonspecific amplification of a primer dimer. In this work, we report an enhancing LAMP technique in terms of assay sensitivity and reliability through streamlined integration with a nonpowered nanoelectric preconcentration (NPP). The NPP, serving as a sample preparation tool, enriched the virus concentration in samples prior to the subsequent LAMP assay. This enrichment enabled not only to achieve more sensitive assay but also to shorten the assay time for all tested clinical samples by ∼10 min compared to the conventional LAMP. The shortened assay time suppresses the occurrence of nonspecific amplification by not providing the necessary incubation time, effectively suppressing misidentification by false positives. Utilizing this technique, we also developed a prototype of the POC NPP-LAMP kit. This kit offers a streamlined diagnostic process for nontrained individuals, from the sample enrichment, transfer of the enriched sample to LAMP assays, which facilitates on-site/on-demand diagnosis of SARS-CoV-2. This development holds the potential to contribute toward preventing not only the current outbreak but also future occurrences of pandemic viruses.</abstract><cop>United States</cop><pub>American Chemical Society</pub><pmid>38393745</pmid><doi>10.1021/acs.analchem.3c05236</doi><tpages>9</tpages><orcidid>https://orcid.org/0000-0001-6002-1557</orcidid></addata></record> |
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| ispartof | Analytical chemistry (Washington), 2024-03, Vol.96 (9), p.3844-3852 |
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| subjects | analytical chemistry Assaying COVID-19 COVID-19 - diagnosis COVID-19 infection Enrichment Gene amplification Humans loop-mediated isothermal amplification Molecular Diagnostic Techniques - methods Nucleic Acid Amplification Techniques - methods pandemic Pandemics point-of-care systems prototypes Reproducibility of Results risk RNA, Viral Sample preparation Sensitivity and Specificity Severe acute respiratory syndrome coronavirus 2 Viral diseases viruses |
| title | Enhancing Loop-Mediated Isothermal Amplification through Nonpowered Nanoelectric Preconcentration |
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