BIOCHIP AND TARGET DNA QUANTITATIVE METHOD

摘要

A biochip used for quantitative analysis of a target DNA contained in a sample. The biochip includes a type I chamber that includes a primer designed to bind to the target DNA, an internal standard DNA of a first amount that has a sequence different from a sequence of the target DNA, and is amplifiable with the primer, and a fluorescent probe that is designed to bind to a part of a PCR product of the target DNA and to a part of a PCR product of the internal standard DNA. The fluorescent probe fluoresces differently for the PCR product of the target DNA and the PCR product of the internal standard DNA. The biochip also includes a type II chamber that includes the internal standard DNA of a second amount, the primer, and the fluorescent probe. The first and second amounts are different.

主权项

1. A quantitative method for a target DNA contained in a biological sample using a biochip, comprising:
introducing a sample into a type I chamber and a type II chamber of the biochip; the type I chamber including;
a primer that is designed to bind to the target DNA;a internal standard DNA of a first amount that has a sequence different from a sequence of the target DNA, and is amplifiable with the primer; anda fluorescent probe that is designed to bind to a part of a PCR product of the target DNA and to apart of a PCR product of the internal standard DNA, and fluoresces differently for the PCR product of the target DNA and the PCR product of the internal standard DNA, and the type II chamber including;
the internal standard DNA of a second amount different from the first amount;the primer; andthe fluorescent probe, performing a PCR amplification in the type I chamber and the type II chamber; measuring a fluorescence intensity of the fluorescent probe in the type I chamber and the type II chamber; determining a regression curve of equation (1) below based on the measured fluorescence intensities in the type I chamber and the type II chamber, and amounts of the internal standard DNA contained in the type I chamber and the type II chamber,
F=a/(C+b)+c  (1)where F is a fluorescence intensity, C is an amount of internal standard DNA, b is an amount of target DNA contained in the sample, and a and c are predetermined values; and
estimating the amount of the target DNA contained in the sample based on the regression curve.