Micro-fluidic chip and portable infrared isothermal PCR platform
A microfluidic chip and infrared technology, applied in the field of warm PCR platform, can solve the problems that hinder patients from seeking medical treatment, achieve the effect of cheap and fast molecular diagnosis selection, improved function, and simple process
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Embodiment 1
[0050] Example 1. Design and preparation of microfluidic chip
[0051] In a specific embodiment, the microfluidic chip (such as figure 1 and 2 Shown) includes a lower cover 18, a MWNT heating layer, a PCR chamber layer, an upper cover 4, an RNA extraction chamber 7, and an RNA extraction chamber cover 6 arranged from bottom to top; the PCR chamber layer is provided with a chamber, and the chamber includes sequential passages The connected hydrogen peroxide chamber 12 , PCR sample chamber 11 and PCR chamber 9 also include a temperature control chamber 10 .
[0052] The MWNT heating layer includes a hydrogen peroxide chamber MWNT heating element 14, a temperature control chamber MWNT heating element 16, and a PCR chamber MWNT heating element corresponding to the hydrogen peroxide chamber 12, the temperature control chamber 10, the PCR chamber 9, and the RNA extraction chamber 7, respectively. 15 and RNA extraction chamber MWNT heating element 13. The RNA extraction chamber 7 ...
Embodiment 2
[0059] Embodiment 2, design of portable infrared isothermal PCR platform
[0060] In a specific example (such as image 3 and 4 As shown in ), the portable infrared isothermal PCR platform consists of three module components: a temperature control module component, a fluorescence detection module component 40 and a positioning module component. The portable infrared isothermal PCR platform is used in conjunction with the above-mentioned microfluidic chip.
[0061] The temperature module components include infrared LED22 (20W, 840nm), thermocouple 21 (TT-K-36-SLE, ETA), thermocouple cold junction compensation circuit and LED circuit; infrared LED22 is electrically connected with LED circuit; thermocouple 21 is connected with thermoelectric The dual cold junction compensation circuit is electrically connected.
[0062] Fluorescence detection module components (such as Figure 5 shown) includes blue LED49 (1400mW, 460nm, SMB490-1100-01, Epitex), bandpass filter 45 (532nm, HB-...
Embodiment 3
[0067] Embodiment 3, sample pretreatment and PCR procedure
[0068] like Figure 11 Mix 0.5 mL of the diluted sample with 0.5 mL of storage buffer as indicated. Add 400 µL of mixed sample and 100 µL of RNA extraction buffer containing magnetic beads to the RNA extraction chamber. Then, the microfluidic chip was placed on the infrared LED, and the sample was heated to 60 °C for 5 min, and then cooled to room temperature for 10 min. Place a magnet under the RNA extraction chamber. After the magnetic beads are absorbed to the bottom, pipette waste from the extraction chamber. Then, the magnet was removed and 1 mL of wash buffer was added to the chamber to wash the beads. Repeat steps 4 and 5 twice. Then, add 40 µL of detection buffer to the RNA extraction chamber and transfer the sample to the PCR sample chamber. After the PCR sample chamber lids were affixed, the hydrogen peroxide chamber was heated to generate oxygen, which in turn drove the samples into the three PCR cha...
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