Two Multiplex PCR Methods for Detecting Several Pathogens Associated with Feline Respiratory and Intestinal Tracts

Area of Science:

• Veterinary Medicine
• Molecular Diagnostics
• Feline Infectious Diseases

Background:

• Feline respiratory and intestinal diseases pose significant health risks.
• Coinfections with multiple pathogens are frequent in clinical feline cases.
• Accurate and rapid diagnosis is crucial for effective treatment.

Purpose of the Study:

• To develop novel multiplex PCR (mPCR) assays for simultaneous detection of key feline pathogens.
• To establish diagnostic tools for both intestinal and respiratory coinfections in cats.

Main Methods:

• Two novel multiplex polymerase chain reactions (mPCRs) were designed and validated.
• Detection of four intestinal pathogens: feline coronavirus (FCoV), feline astrovirus (FeAstV), feline panleukopenia virus (FPV), and feline kobuvirus (FeKoV).
• Detection of five respiratory pathogens: feline calicivirus (FCV), feline herpesvirus 1 (FHV-1), feline leukemia virus (FeLV), *Chlamydia felis* (*C. felis*), and influenza A virus (IAV).

Main Results:

• Sensitivity analysis determined detection limits ranging from 10^3 to 10^5 copies/µL for the targeted pathogens.
• Both mPCR assays demonstrated high specificity.
• The mPCR methods performed effectively when applied to clinical feline samples.

Conclusions:

• Two novel mPCR methods were successfully established for feline pathogen detection.
• These assays provide a valuable tool for the simultaneous diagnosis and monitoring of feline respiratory and intestinal coinfections.
• The developed methods can aid in timely and effective clinical management of feline infectious diseases.