Loop-mediated isothermal amplification integrated on microfluidic chips for point-of-care quantitative detection of pathogens

Anal Chem. 2010 Apr 1;82(7):3002-6. doi: 10.1021/ac1000652.

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 (microLAMP). 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 microL 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 degrees C). A real time quantitative microLAMP assay using absorbance detection is possible by integration of optical fibers within the chip.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • DNA, Viral / analysis*
  • Diphosphates / chemistry
  • Herpesvirus 1, Suid / genetics
  • Microfluidic Analytical Techniques / methods*
  • Nucleic Acid Amplification Techniques / methods*
  • Point-of-Care Systems

Substances

  • DNA, Viral
  • Diphosphates