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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

Pending Publication Date: 2021-07-02
深圳市刚竹医疗科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The throughput of the traditional fluorescent PCR method is low, and basically only one pathogen can be detected

Method used

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  • Nucleic acid combination product, detection kit and micro-fluidic chip
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  • Nucleic acid combination product, detection kit and micro-fluidic chip

Examples

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Effect test

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

The invention relates to a nucleic acid combination product, a detection kit and a micro-fluidic chip. The nucleic acid combination product comprises at least two of the following detection primer pairs: a feline parvovirus primer pair, a feline coronavirus primer pair, a feline herpesvirus type I primer pair, a feline calicivirus primer pair, a feline leukemia virus primer pair, a feline immunodeficiency virus primer pair, a feline astrovirus primer pair, a feline rotavirus primer pair, a cat vaccinia virus primer pair and a cat reovirus primer pair. The nucleic acid combination product can be used for detecting at least two feline pathogenic viruses in the sample to be detected at one time, and is good in specificity and relatively high in detection sensitivity.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a nucleic acid combination product, a detection kit and a microfluidic chip. Background technique [0002] With the improvement of socioeconomic level and human spiritual level, more and more people keep pet cats. It is understood that the number of pet dogs and cats in cities and towns across the country reached 99.15 million in 2019, an increase of 7.66 million over 2018; among them, the number of pet cats was 44.12 million, an increase of 8.6% over 2018. [0003] However, the current level of disease detection technology in the pet medical industry is generally low, and the frequent and new cases of cats are also on the rise year by year. Common feline infectious viruses include feline parvovirus, feline coronavirus, feline herpesvirus type I, Feline calicivirus, feline leukemia virus, feline immunodeficiency virus, feline astrovirus, feline rotavirus, feline vaccinia virus, feli...

Claims

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Application Information

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IPC IPC(8): C12Q1/70C12Q1/686C12Q1/6806C12N15/11C12M1/34C12M1/00B01L3/00C12R1/93
CPCC12Q1/701C12Q1/705C12Q1/702C12Q1/703C12Q1/686C12Q1/6806B01L3/5027C12Q2600/166B01L2200/10B01L2300/0861C12Q2521/107C12Q2565/629C12Q2563/107C12Q2545/101C12Q2527/125
Inventor 游腾飞
Owner 深圳市刚竹医疗科技有限公司
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