مقالات پذیرفته شده در نهمین کنگره بین المللی زیست پزشکی
Design and production of molecular detection kit (RT-PCR) for feline FIP virus through specific primer design
Design and production of molecular detection kit (RT-PCR) for feline FIP virus through specific primer design
Maryam Rahmani,1,*Shaghayegh Mehdizadeh,2
1. department of Biology,East Tehran Branch, Islamic Azad University, Tehran, Iran 2. Department of Biology, East Tehran Branch, Islamic Azad University, Tehran, Iran
Introduction: Feline Infectious Peritonitis (FIP) is one of the most fatal viral diseases in cats. It is caused by a strain of feline coronavirus (FCoV) that mutates to produce a systemic pathogenic form called FIPV . FCoV is asymptomatic or mild in most cats, but in some cases, the virus mutates in the host to form FIP, which leads to generalized inflammation, fluid accumulation in the abdomen or chest, and death of the animal.
Diagnosis of FIP is difficult due to clinical similarities with other diseases and usually requires invasive and complex procedures.
Methods: 1.Design specific primers for the detection of FIP virus using target gene sequences.
2. Evaluate the accuracy, sensitivity, and specificity of the designed primer.
3. Produce a prototype molecular detection kit (proto-kit) for use in veterinary clinics
Results: 1.Genetic data extraction: Extraction of FIP virus genomic sequences from the NCBI GenBank database and identification of target regions.
2. Primer design: Use of Primer3, Oligo7, and Primer-BLAST software to design specific primers.
3. Sampling: Collection of samples from suspected cats and extraction of viral RNA.
4. RT-PCR: Synthesis of cDNA and PCR reaction with designed primers.
5. Analysis of results: Evaluation of PCR bands by electrophoresis and comparison with control samples
Conclusion: Due to the lack of rapid and non-invasive methods for definitive diagnosis of FIP, sick cats often remain untreated or are diagnosed late. The development of a molecular diagnostic kit can be of great help to veterinarians, animal clinics, and even animal welfare organizations . On the other hand, with the spread of coronaviruses in different species, research on related viruses such as FCoV is also of epidemiological importance.