Loop-Mediated Isothermal Amplification Integrated on Microfluidic Chips for Point-of-Care Quantitative Detection of Pathogens
This work shows that loop-mediated isothermal amplification (LAMP) of nucleic acid can be integrated in an eight-channel microfluidic chip for readout either by the naked eye (as a result of the insoluble byproduct pyrophosphate generating during LAMP amplification) or via absorbance measured by an...
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| Veröffentlicht in: | Analytical chemistry (Washington) 2010-04, Vol.82 (7), p.3002-3006 |
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| container_title | Analytical chemistry (Washington) |
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| creator | Fang, Xueen Liu, Yingyi Kong, Jilie Jiang, Xingyu |
| description | This work shows that loop-mediated isothermal amplification (LAMP) of nucleic acid can be integrated in an eight-channel microfluidic chip for readout either by the naked eye (as a result of the insoluble byproduct pyrophosphate generating during LAMP amplification) or via absorbance measured by an optic sensor; we call this system microLAMP (μLAMP). It is capable of analyzing target nucleic acids quantitatively with high sensitivity and specificity. The assay is straightforward in manipulation. It requires a sample volume of 0.4 μL and is complete within 1 h. The sensitivity of the assay is comparable to standard methods, where 10 fg of DNA sample could be detected under isothermal conditions (63 °C). A real time quantitative μLAMP assay using absorbance detection is possible by integration of optical fibers within the chip. |
| doi_str_mv | 10.1021/ac1000652 |
| format | Article |
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It is capable of analyzing target nucleic acids quantitatively with high sensitivity and specificity. The assay is straightforward in manipulation. It requires a sample volume of 0.4 μL and is complete within 1 h. The sensitivity of the assay is comparable to standard methods, where 10 fg of DNA sample could be detected under isothermal conditions (63 °C). 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Chem</addtitle><description>This work shows that loop-mediated isothermal amplification (LAMP) of nucleic acid can be integrated in an eight-channel microfluidic chip for readout either by the naked eye (as a result of the insoluble byproduct pyrophosphate generating during LAMP amplification) or via absorbance measured by an optic sensor; we call this system microLAMP (μLAMP). It is capable of analyzing target nucleic acids quantitatively with high sensitivity and specificity. The assay is straightforward in manipulation. It requires a sample volume of 0.4 μL and is complete within 1 h. The sensitivity of the assay is comparable to standard methods, where 10 fg of DNA sample could be detected under isothermal conditions (63 °C). A real time quantitative μLAMP assay using absorbance detection is possible by integration of optical fibers within the chip.</description><subject>Analytical chemistry</subject><subject>Chemistry</subject><subject>Deoxyribonucleic acid</subject><subject>Diphosphates - chemistry</subject><subject>DNA</subject><subject>DNA, Viral - analysis</subject><subject>Exact sciences and technology</subject><subject>Fiber optics</subject><subject>General, instrumentation</subject><subject>Herpesvirus 1, Suid - genetics</subject><subject>Integration</subject><subject>Microfluidic Analytical Techniques - methods</subject><subject>Nucleic Acid Amplification Techniques - methods</subject><subject>Point-of-Care Systems</subject><subject>Real time</subject><subject>Spectrometric and optical methods</subject><issn>0003-2700</issn><issn>1520-6882</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2010</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNplkUuLFDEQx4Mo7jh68AtIEBbx0JpX53FcxtfALK6g5yaTruxk6e60SVrw4Hc37o47oKeiil_96_FH6Dklbyhh9K11lBAiW_YArWjLSCO1Zg_RqhZ5wxQhZ-hJzjeEUEqofIzOWO3SrWIr9GsX49xcQh9sgR5vcywHSKMd8MU4D8EHZ0uIE95OBa7TLVOzy-BS9MMS-uDw5hDmjH1M-CqGqTTRNxubAH9Z7FRCqf0_AL-DAu5WKXp8ZcshXsOUn6JH3g4Znh3jGn378P7r5lOz-_xxu7nYNVZwVRpnjCPM25bLXhuhesPrscRRqhSXxoElZs9VKwSRGsD3Qu_BGWVE31JQnq_RqzvdOcXvC-TSjSE7GAY7QVxypzjn2kglKvnyH_ImLmmqy3WMKq1lW0eu0es7qH4h5wS-m1MYbfrZUdL9caS7d6SyL46Cy36E_p78a0EFzo-Azc4OPtnJhXziWKuMFOLEWZdPS_0_8DfS1J5D</recordid><startdate>20100401</startdate><enddate>20100401</enddate><creator>Fang, Xueen</creator><creator>Liu, Yingyi</creator><creator>Kong, Jilie</creator><creator>Jiang, Xingyu</creator><general>American Chemical Society</general><scope>IQODW</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>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></search><sort><creationdate>20100401</creationdate><title>Loop-Mediated Isothermal Amplification Integrated on Microfluidic Chips for Point-of-Care Quantitative Detection of Pathogens</title><author>Fang, Xueen ; Liu, Yingyi ; Kong, Jilie ; Jiang, Xingyu</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-a437t-c99c02fa536d8947d936520c1177369cea09b37544068eefd48bec9794d51e7f3</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2010</creationdate><topic>Analytical chemistry</topic><topic>Chemistry</topic><topic>Deoxyribonucleic acid</topic><topic>Diphosphates - chemistry</topic><topic>DNA</topic><topic>DNA, Viral - analysis</topic><topic>Exact sciences and technology</topic><topic>Fiber optics</topic><topic>General, instrumentation</topic><topic>Herpesvirus 1, Suid - genetics</topic><topic>Integration</topic><topic>Microfluidic Analytical Techniques - methods</topic><topic>Nucleic Acid Amplification Techniques - methods</topic><topic>Point-of-Care Systems</topic><topic>Real time</topic><topic>Spectrometric and optical methods</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Fang, Xueen</creatorcontrib><creatorcontrib>Liu, Yingyi</creatorcontrib><creatorcontrib>Kong, Jilie</creatorcontrib><creatorcontrib>Jiang, Xingyu</creatorcontrib><collection>Pascal-Francis</collection><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><jtitle>Analytical chemistry (Washington)</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Fang, Xueen</au><au>Liu, Yingyi</au><au>Kong, Jilie</au><au>Jiang, Xingyu</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Loop-Mediated Isothermal Amplification Integrated on Microfluidic Chips for Point-of-Care Quantitative Detection of Pathogens</atitle><jtitle>Analytical chemistry (Washington)</jtitle><addtitle>Anal. Chem</addtitle><date>2010-04-01</date><risdate>2010</risdate><volume>82</volume><issue>7</issue><spage>3002</spage><epage>3006</epage><pages>3002-3006</pages><issn>0003-2700</issn><eissn>1520-6882</eissn><coden>ANCHAM</coden><abstract>This work shows that loop-mediated isothermal amplification (LAMP) of nucleic acid can be integrated in an eight-channel microfluidic chip for readout either by the naked eye (as a result of the insoluble byproduct pyrophosphate generating during LAMP amplification) or via absorbance measured by an optic sensor; we call this system microLAMP (μLAMP). It is capable of analyzing target nucleic acids quantitatively with high sensitivity and specificity. The assay is straightforward in manipulation. It requires a sample volume of 0.4 μL and is complete within 1 h. The sensitivity of the assay is comparable to standard methods, where 10 fg of DNA sample could be detected under isothermal conditions (63 °C). A real time quantitative μLAMP assay using absorbance detection is possible by integration of optical fibers within the chip.</abstract><cop>Washington, DC</cop><pub>American Chemical Society</pub><pmid>20218572</pmid><doi>10.1021/ac1000652</doi><tpages>5</tpages></addata></record> |
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| subjects | Analytical chemistry Chemistry Deoxyribonucleic acid Diphosphates - chemistry DNA DNA, Viral - analysis Exact sciences and technology Fiber optics General, instrumentation Herpesvirus 1, Suid - genetics Integration Microfluidic Analytical Techniques - methods Nucleic Acid Amplification Techniques - methods Point-of-Care Systems Real time Spectrometric and optical methods |
| title | Loop-Mediated Isothermal Amplification Integrated on Microfluidic Chips for Point-of-Care Quantitative Detection of Pathogens |
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