Sensing apparatus and method
A method of observing reaction intermediaries during a chemical reaction and comprising detecting an electrical signal output from an ion sensitive field effect transistor exposed to said reaction, and monitoring the detected electrical signal to discriminate discrete fluctuations in the electrical signal, the discrete fluctuations indicating reaction intermediaries occurring during a chemical reaction.
1. A sensing apparatus comprising at least two ion sensitive field effect transistors (ISFET) and at least two sets of reagents;
a first of the transistors exposable to an unknown sample and a first of the sets of reagents, and separated from a second of the transistors exposable to an unknown sample and a second of the sets of reagents, different from the first set of reagents; and
further comprising means for determining the difference between electrical signals generated by the first and second transistors, to provide an output signals to identify the sample.
2. The sensing apparatus according to claim 1 , further comprising microfluidic chambers having a volume of less than 50 μL.
3. The sensing apparatus according to claim 2 , wherein each ISFET is housed within the microfluidic chamber to receive the unknown sample and one of the sets of reagents.
4. The sensing apparatus according to claim 1 , further comprising microchannels to deliver the unknown sample.
5. The sensing apparatus according to claim 4 , further comprising a plate covering a silicon chip integrating the ISFETs, the plate having portions removed to provide said microfluidic chambers.
6. The sensing apparatus according to claim 5 , further comprising a heating element and a temperature sensor integrated into the silicon chip.
7. The sensing apparatus according to claim 2 , further comprising means to deliver the sample to more than one of said microfluidic chambers for simultaneous testing of more than one variation of the sample.
8. The sensing apparatus according to claim 2 , further comprising a seal for the microfluidic chambers.
9. The sensing apparatus according to claim 1 , wherein the unknown sample comprises a target nucleic acid.
10. The sensing apparatus according to claim 1 , wherein the sets of reagents comprise allele specific reagents.
11. The sensing apparatus according to claim 9 , wherein the sets of reagents comprise one or more types of nucleotide and an oligonucleotide serving as a primer or probe for the target nucleic acid.
12. The sensing apparatus according to claim 9 , wherein one of the sets of reagents is complementary and one of the sets of reagents is non-complementary with the target nucleic acid.
13. The sensing apparatus according to claim 1 , wherein each set of reagents comprise magnesium.
14. The sensing apparatus according to claim 1 , further comprising beads having probes or primers immobilised on the surface.
15. The sensing apparatus according to claim 1 , wherein the apparatus further comprises monitoring means arranged to monitor the time at which fluctuations occur in the electrical signals generated by the first and second transistors and the magnitude of said fluctuations.
16. The sensing apparatus according to claim 1 , wherein the first and second transistors are connected to first and second operational amplifiers respectively.
17. The sensing apparatus according to claim 1 , wherein the first and second transistors are arranged to form part of first and second operational amplifiers respectively.
18. The sensing apparatus according to claim 1 , wherein an electrical signal is passed to an electrode to maintain voltages applied to the first and second transistors at a constant level.