Nucleic acid combination product, detection kit and micro-fluidic chip
A microfluidic chip, combined product technology, applied in the biological field, can solve problems such as low throughput
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preparation example Construction
[0071] In addition, an embodiment of the present invention also provides a preparation method of the above-mentioned microfluidic chip, the preparation method includes preparing the PCR reaction reagent, the internal standard, the above-mentioned detection primer pair and the detection probe corresponding to the detection primer pair into a freeze-dried powder step.
[0072] Specifically, the PCR reaction reagent, the internal standard, the above-mentioned detection primer pair and the detection probe were respectively frozen and stored at -80° C. for 8 h to 10 h. Then vacuum freeze-drying is carried out, and after the freeze-drying is completed, the corresponding reagents are placed in the chamber of the microfluidic chip and packaged. Concrete vacuum freeze-drying process parameter is as follows table 1:
[0073] Table 1
[0074]
[0075] The PCR reaction reagent, the internal standard, the detection primer pair and the detection probe after freeze-drying according to t...
Embodiment 1
[0086] The structure of the microfluidic unit of the microfluidic chip of Example 1 is as follows figure 1 shown. The microfluidic unit includes a sample chamber, a lysate storage chamber, a lysis chamber, a cleaning liquid storage chamber, a buffer chamber, an eluent storage chamber, a selection chamber, a nucleic acid chamber, a reaction chamber and a waste liquid chamber. In the microfluidic unit:
[0087] An internal standard is set in the sample cavity, and the internal standard is freeze-dried powder. The internal standard consists of an internal standard primer pair and an internal standard probe, and the specific sequences of the internal standard primer pair and internal standard probe are shown in Table 2;
[0088] The lysate is stored in the lysate storage cavity, and the lysate is composed of guanidine hydrochloride with a final concentration of 5mol / L, C with a final volume percentage of 40%. 2 h 5 OH, Tris-HCl with a final concentration of 40mmol / L and Triton...
Embodiment 2
[0098] (1) Collect 10 positive samples of feline pathogenic virus and 8 negative samples, numbered 1 to 18. Sample No. 1 is a positive sample of feline parvovirus, sample No. 2 is a positive sample of feline coronavirus, and sample No. 3 is a cat Herpes virus type I positive sample, sample No. 4 is a positive sample of feline calicivirus, sample No. 5 is a positive sample of feline leukemia virus, sample No. 6 is a positive sample of feline immunodeficiency virus, and sample No. 7 is a positive sample of feline astrovirus, Sample No. 8 was positive for cat rotavirus, sample No. 9 was positive for feline vaccinia virus, sample No. 10 was positive for feline reovirus, sample No. 11 was positive for canine parainfluenza virus, and sample No. 12 was positive for rotavirus. Sample No. 13 was Chlamydia, Sample No. 14 was Mycoplasma, Sample No. 15 was Canine Adenovirus Type I, Sample No. 16 was Canine Adenovirus Type II, Sample No. 17 was Canine Distemper Virus, and Sample No. 18 was ...
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Abstract
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