Department of Chemistry and Institutes of Biomedical Sciences, Fudan University, Shanghai 200433, P.R. China.
Anal Chem. 2010 Apr 1;82(7):3002-6. doi: 10.1021/ac1000652.
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.
本工作表明,核酸环介导等温扩增(LAMP)可以集成在八通道微流控芯片中,通过肉眼(由于 LAMP 扩增过程中产生的不溶性副产物焦磷酸)或通过光传感器测量的吸光度进行读取;我们称这个系统为 microLAMP(microLAMP)。它能够以高灵敏度和特异性定量分析靶核酸。该测定方法操作简单。它需要 0.4 μL 的样品体积,在 1 小时内完成。该测定的灵敏度与标准方法相当,在等温条件(63°C)下可以检测到 10 fg 的 DNA 样品。通过在芯片中集成光纤,可以实现使用吸光度检测的实时定量 microLAMP 测定。