Abstract: Systems and methods for isothermal amplification of nucleic acids in a ratiometric manner are disclosed. Methods of isothermal amplification of nucleic acids are convenient for areas that lack reliable electricity and/or funding for precision lab equipment. However, these methods typically result in non-ratiometric amplification of target sequences, thus preventing quantitative analysis or application of the results, such as for diagnostic testing. Systems and methods herein provide a method of isothermal amplification that ratiometrically amplifies target sequences, thus expanding the reach of diagnostics into remote and/or economically disadvantaged areas.