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 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.
Fang
Xueen
X
Department of Chemistry and Institutes of Biomedical Sciences, Fudan University, Shanghai 200433, P.R. China.
Liu
Yingyi
Y
Kong
Jilie
J
Jiang
Xingyu
X
D004279:DNA, Viral / Q000032:analysis*; D011756:Diphosphates / Q000737:chemistry; D011558:Herpesvirus 1, Suid / Q000235:genetics; D046210:Microfluidic Analytical Techniques / Q000379:methods*; D021141:Nucleic Acid Amplification Techniques / Q000379:methods*; D019095:Point-of-Care Systems
D004279:DNA, Viral; D011756:Diphosphates