Primer group, product and method for detecting feline respiratory tract pathogens as well as application

A technique of respiratory tract and primer set, applied in the field of nucleic acid detection, can solve the problems of increasing the difficulty of diagnosis, aggravating the cat's condition, mixed infection, etc., and achieve the effect of reducing the cost and time of detection, good specificity and high sensitivity of the results.

Inactive Publication Date: 2020-07-28
HANGZHOU BIOER TECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0007] The pathogens of the above five kinds of cats can all belong to respiratory pathogens, and the clinical symptoms are mostly similar, and even mixed infections are often found, which increases the difficulty of diagnosis and increases the risk of cats getting worse without symptomatic treatment. Therefore, it was detected and identified The type of pathogen is particularly important

Method used

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  • Primer group, product and method for detecting feline respiratory tract pathogens as well as application
  • Primer group, product and method for detecting feline respiratory tract pathogens as well as application
  • Primer group, product and method for detecting feline respiratory tract pathogens as well as application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0102] 1 primer probe

[0103] 1.1 Design and screening of primers and probes

[0104] The inventor downloaded a large number of gene sequences of feline herpesvirus type 1, feline calicivirus, mycoplasma, feline chlamydia and bronchiseptica Bordetella from GenBank, and found the highly conserved regions of the above five pathogens through sequence comparison. Primer probes, at least two sets of primers and one probe are designed for each pathogen, and the performance of the primer probes is verified through experimental comparisons, and primer sets, probes and their combinations suitable for multiple detection of feline respiratory pathogens are screened out. The sequence is as follows :

[0105] SEQ ID NO: 1: 5'-CAAGTATGACGGCTTACGATC-3';

[0106] SEQ ID NO: 2: 5'-GTTGGGTCTTATGTGCCAGTC-3';

[0107] SEQ ID NO: 3: 5'-TCAGCGTTGTTGTCACAGTTCTA-3';

[0108] SEQ ID NO: 4: 5'-GCTTAAGATACCCAATCTCGTA-3';

[0109] SEQ ID NO: 5: 5'-GCTTACAGCTATTAGTTCACT-3';

[0110] SEQ ID NO: 6: 5...

Embodiment 2

[0150] Example 2 Detection of Cat Respiratory Tract Samples Collected in Pet Hospital

[0151] 1 Experimental operation

[0152] 1.1 Sample source

[0153] A pet hospital in a certain place has a cat disease detection room, and some samples are now collected for detection of respiratory pathogens.

[0154] 1.2 Sample processing

[0155] Use the Simply P virus DNA / RNA extraction kit from Hangzhou Bioer Technology Co., Ltd. to extract nucleic acid from the sample, and the extraction steps refer to the kit instructions.

[0156] 1.3 Amplification reagent preparation

[0157] According to the number of samples to be tested, take out the corresponding number of PCR tubes containing the fluorescent PCR reaction solution, and use a hand-held centrifuge to centrifuge instantaneously after melting, so that the reagents are concentrated at the bottom of the tube. Open the cap of the PCR tube, add 5 μL each of the negative and positive control substance, the positive control substanc...

Embodiment 3

[0171] Embodiment 3 specificity verification experiment

[0172] Feline herpesvirus type 1, feline calicivirus, mycoplasma, feline chlamydia, bronchiseptic Bordetella, canine herpes virus, feline parvovirus, feline coronavirus, toxoplasma gondii according to the feline respiratory pathogen detection method described in embodiment 1 , Mycoplasma hemophilus, positive plasmid pFHV-1, positive plasmid pFCV, positive plasmid pMyc, positive plasmid pF-Chl, positive plasmid pBb, positive control substance, negative control substance, test and analyze the results, negative and positive control substances meet the requirements, Therefore, it shows that the experiment is valid. Feline herpesvirus type 1 and positive plasmid pFHV-1 only have Ct values ​​for the FAM channel, feline calicivirus and positive plasmid pFCV only have Ct values ​​for the HEX channel, mycoplasma and positive plasmid pMyc only have Ct values ​​for the ROX channel, feline Chlamydia and positive plasmids have Ct va...

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Abstract

The invention provides a primer, product and method for detecting feline respiratory tract pathogens as well as application, and relates to the technical field of nucleic acid tests. In the primer group for detecting the feline respiratory tract pathogens, a primer for detecting feline herpesvirus type 1 (FHV1), a primer for detecting feline calicivirus (FCV), a primer for detecting mycoplasma, aprimer for detecting feline chlamydia and a primer for detecting bordetella bronchiseptica are primers designed through highly conservative regions of each pathogen, have no cross reaction with otherfeline pathogens, and have the characteristics of high specificity and high sensitivity. By use of the primer group, five feline respiratory tract pathogens including the FHV1, the FCV, the mycoplasma, the feline chlamydia and the bordetella bronchiseptica can be identified in the same reaction system, and the detection cost and time are greatly reduced as compared with a traditional method.

Description

technical field [0001] The invention relates to the technical field of nucleic acid detection, in particular to a primer set, product, method and application for detecting feline respiratory pathogens. Background technique [0002] Feline herpesvirus type 1 (Feline herpesvirus type 1, FHV1) is the causative agent of feline viral rhinotracheitis, belonging to the αherpesvirinae and varicella genus in the family Herpesviridae, and can cause acute and highly contact upper respiratory tract diseases in felines . Young cats have weak resistance to the virus, with a high morbidity rate and a fatality rate of about 50%. It is currently known as one of the most important cat respiratory diseases. [0003] Feline Calicivirus (Feline Calicivirus, FCV) belongs to the Feline Caliciviridae family, a member of the Calicivirus genus, and is a single-stranded RNA with no envelope and a spherical capsid (there are multiple cup-shaped depressions on the capsid) The virus is highly contagiou...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/70C12Q1/689C12Q1/686C12Q1/04C12N15/11C12R1/35C12R1/01
CPCC12Q1/686C12Q1/689C12Q1/701C12Q1/705C12Q2600/16C12Q2563/107C12Q2545/114
Inventor 田星陈芝娟王虹军贺贤汉
Owner HANGZHOU BIOER TECH CO LTD
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