What is Canine Adenovirus Type 1 (CAV-1) Fluorescence Quantification Test Kit?

I. Definition

The Canine Adenovirus Type 1 (CAV – 1) real – time fluorescence quantitative PCR detection kit is a specialized tool for detecting Canine Adenovirus Type 1. It employs real – time fluorescence quantitative PCR (polymerase chain reaction) technology and is capable of rapidly and accurately quantitatively detecting the nucleic acid of Canine Adenovirus Type 1 in samples.

II. Detection Principle

1. Principle of real – time fluorescence quantitative PCR
   • This kit is based on real – time fluorescence quantitative PCR technology. Primers and fluorescence probes designed for the specific gene sequences of Canine Adenovirus Type 1 are added to the reaction system.
   • When Canine Adenovirus Type 1 is present in the sample, the primers will specifically bind to the corresponding sequences of the viral nucleic acid. Under the action of DNA polymerase, chain elongation occurs according to the principle of base – pair complementarity, with dNTP (deoxyribonucleoside triphosphate) as the raw material.
   • The fluorescence probe does not emit fluorescence in its intact state. As the PCR reaction proceeds, Taq enzyme (a type of DNA polymerase) will cut the fluorescence probe during the elongation process, separating the fluorescent reporter group from the quenching group and thus releasing a fluorescence signal.
   • With each PCR cycle, there is a corresponding increase in the fluorescence signal. By detecting the change in fluorescence signal intensity with the number of cycles and combining with the standard curve established from known standards, quantitative analysis of the nucleic acid content of Canine Adenovirus Type 1 in the sample can be carried out.

III. Kit Components

1. Reaction buffer
   • Its function is to provide a stable chemical environment for the PCR reaction, including maintaining appropriate pH values and ion concentrations, etc., ensuring that enzymes such as DNA polymerase can function normally.
2. Primers and fluorescence probes
   • The primers are short – chain DNA fragments designed according to the specific gene sequences of Canine Adenovirus Type 1 and can guide DNA synthesis. The fluorescence probe is an oligonucleotide chain with a fluorescent reporter group and a quenching group and is used to detect the amplification of viral nucleic acid.
3. DNA polymerase
   • Usually Taq enzyme, it is a key enzyme for catalyzing the elongation of DNA strands in the PCR reaction. Based on the primers, it can synthesize new DNA strands with dNTP as the raw material.
4. dNTP
   • It is the raw material for synthesizing new DNA strands, including dATP (deoxyadenosine triphosphate), dCTP (deoxycytidine triphosphate), dGTP (deoxyguanosine triphosphate), and dTTP (deoxyuridine triphosphate), providing the material basis for the synthesis of DNA strands in the PCR reaction.
5. Positive and negative controls
   • The positive control is a sample containing a known quantity of Canine Adenovirus Type 1 nucleic acid and is used to verify the effectiveness of the kit and the accuracy of the detection process. The negative control is a sample without Canine Adenovirus Type 1 nucleic acid and is used to exclude false – positive results, ensuring the reliability of the test results.

IV. Types of Test Samples

1. Clinical samples
   • Mainly including blood, urine, eye secretions, and nasal secretions of dogs. For example, collecting blood or eye secretion samples from dogs with suspected Canine Adenovirus Type 1 infection symptoms such as corneal opacity and fever for testing is helpful for disease diagnosis.
2. Environmental samples
   • For example, environmental swabs from places such as dog kennels and pet hospitals. Collecting these environmental samples for testing can help understand the presence of Canine Adenovirus Type 1 in the environment and provide a basis for preventing and controlling virus transmission.

V. Detection Steps

1. Sample collection and processing
   • Samples are collected in accordance with the specifications of sample types. When collecting clinical samples, attention should be paid to avoiding sample contamination. Collected samples may need pre – treatment. For example, blood samples may need to be centrifuged to separate serum or plasma, and some secretion samples may need to be diluted or filtered.
2. Nucleic acid extraction
   • Appropriate nucleic acid extraction methods are used to extract the nucleic acid of Canine Adenovirus Type 1 from the processed samples. Common nucleic acid extraction methods include column – based extraction and magnetic bead extraction, etc. The purpose is to obtain pure viral nucleic acid for subsequent testing.
3. Preparation of the reaction system
   • According to the requirements of the kit instructions, accurately mix components such as reaction buffer, primers, fluorescence probes, DNA polymerase, dNTP, and extracted nucleic acid samples (or control samples) to prepare the PCR reaction system.
4. PCR reaction
   • Place the prepared reaction system into a real – time fluorescence quantitative PCR instrument, set appropriate reaction programs, including temperature and time parameters for steps such as pre – denaturation, denaturation, annealing, and elongation, and start the PCR reaction.
5. Result analysis
   • Analyze the curve of the change in fluorescence signal intensity with the number of cycles through the software of the real – time fluorescence quantitative PCR instrument. Calculate the nucleic acid content of Canine Adenovirus Type 1 in the sample based on the standard curve established by the positive control, and judge whether the sample is infected with Canine Adenovirus Type 1 and the degree of infection accordingly.

VI. Application Fields

1. Veterinary clinical diagnosis
   • It helps veterinarians quickly and accurately diagnose whether dogs are infected with Canine Adenovirus Type 1 so that treatment plans can be formulated in a timely manner. For example, for dogs with symptoms such as hepatitis and respiratory diseases, testing with this kit can determine whether the cause is Canine Adenovirus Type 1 infection, enabling targeted treatment.
2. Disease monitoring and prevention and control
   • In places such as dog breeding farms, pet hospitals, and animal shelters, it can be used for disease monitoring of dog populations. Through regular testing, potential infected cases can be detected in a timely manner, and prevention and control measures such as isolation and disinfection can be taken to prevent the spread of Canine Adenovirus Type 1 among dog populations.
3. Quarantine inspection for dog import and export
   • In the process of dog import and export, this kit can be used for quarantine inspection to ensure that imported and exported dogs do not carry Canine Adenovirus Type 1, ensuring the animal health and safety of countries and regions.