Microfluidic chip for multiple loop-mediated isothermal amplification (LAMP) detection and preparation method thereof
A microfluidic chip and chip technology, which is applied in the field of temperature amplification, can solve the problem of not being able to detect multiple targets at the same time, and achieve the effects of being conducive to popularization and application, satisfying effective and rapid diagnosis, and intuitive results.
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Embodiment 1
[0027] Example 1 Rapid typing detection of influenza A virus (Flu A virus), seasonal influenza A virus (Seasonal flu A virus) and 2009 swine-derived influenza A virus (Pandemic flu A virus).
[0028] The specific design size of the chip: the size of the amplification pool is (length: 10 mm, width: 0.6 mm, depth: 0.8 mm); the size of the capillary channel is (length: 2 mm, width: 0.1 mm, depth: 0.1 mm).
[0029] The specific LAMP probes of the three influenza viruses were respectively coated on the 1st-4th amplification pool 1(1'), 2(2'), 3(3'); the amplification pool 4(4') A positive control LAMP probe was applied; the fifth amplification pool 5 (5') was not coated with a probe, and served as a blank control. We use this functional chip to detect and type three important influenza viruses.
[0030] Take 4 μL of the three virus standard solutions and clinical reference products respectively, add them to the sample wells of the chip, then add 46 μL of LAMP amplification solutio...
Embodiment 2
[0036] Embodiment 2, the typing detection of eight kinds of important swine disease viruses
[0037] The specific size of the chip is the same as above, LAMP multi-chip amplification pools 1-8 are respectively coated for porcine foot-and-mouth disease virus (FMDV), swine fever virus (CSFV), porcine blue ear disease virus (PRRSV), porcine transmissible gastroenteritis Virus (TGEV), porcine pseudorabies virus (PRV), porcine parvovirus (PPV), porcine circovirus (PCV), swine influenza virus (SIV) LAMP-specific nucleic acid probes for eight important porcine disease viruses, forming an eight Functional typing chip of swine disease virus; No. 9 amplified pool was coated with other probes and used as a negative control; No. 10 amplified pool was not coated with probes and used as a blank control.
[0038] Add sample 1 (FMDV), sample 2 (CSFV), sample 3 (PRRSV), sample 4 (TGEV), sample 5 (PRV), sample 6 (PPV), sample 7 (PCV), Sample 8 (SIV), Sample 9 (FMDV, CSFV), Sample 10 (FMDV, CSF...
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