Team:USTC/Hardware

USTC-iGEM-Hardware


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Background


     Recent studies have shown that the accumulation of tau protein in peripheral blood of patients with Alzheimer's disease is closely related to the early stage of Alzheimer's disease, which makes the monitoring of tau protein in blood very important. At present, the detection of trace tau protein in patients' blood can only rely on high-cost equipment and professional training, which makes it more difficult to obtain the universality of tau protein detection. Therefore, we designed this hardware to measure the amount of tau protein in blood in an easier way. No special training is required to operate the device, and we want to make it easier for the public to detect tau in their blood so they can predict and prevent Alzheimer's disease.

Simple qPCR overview




     The core of the device is an easy-to-use, user-friendly, customizable biomarker detection device. Different from traditional biological detection devices, our signal amplification adopts RPA amplification, that is, isothermal amplification. Therefore, the whole device forms a constant temperature air bath under the action of heater and special aluminum shell to ensure that the whole reaction is carried out at isothermal temperature. The signal was amplified by a simple aluminum QPCR device heated by air bath and fluorescence detection was carried out in real time. Finally, the content of related biomarkers was calculated by equation.

Function introduction




     As for the heating of the equipment, the very bottom of the equipment is placed with a very sensitive temperature control device, through four heating plates to heat the middle of the silver aluminum air bath. Through understanding and analysis, the heating device can make the reaction system temperature at the set temperature for 30 minutes or more, which is enough to meet our entire reaction.


     About signal detection, because of we adopt is FAM fluorescent probe, we can produce according to the probe fluorescence intensity, and we can measure the concentration range, select the appropriate light sensor (PP730), placed at the top of the equipment, dedicated to detect EP tube samples FAM probe fluorescence intensity, thus fluorescent signal into the computer, The next step is fitting to get the data we want. The fluorescence signal is activated by the blue LED light below to the probe in the EP tube. In order to prevent interference from other wavelengths of light, two 470nm filters (HS-470ZDLGP-Y19.5) are used so that only 470nm light passes through and activates the FAM fluorescence. After that, the fluorescence passes through the orange filter (HS-CB535-Y16) and finally reaches the photosensitive sensor on the top to obtain the desired fluorescence signal.
     As for the operation of the equipment, remove the photosensitive sensor through the upper mechanical arm, place the prepared EP tube (containing plasma and reagents) to the central support of the aluminum air pot, and then install the photosensitive sensor to obtain a dark environment with good tightness, which is conducive to the detection of fluorescence signal. After setting the temperature required by the reaction (39℃), start the set program for detection. The light-sensitive sensor will measure the fluorescent signal at regular intervals, and a computer connected to it will record the data in real time.