Key result
The TaqMan qRT-PCR assay detected feline calicivirus in 21% of clinical samples compared to only 5% using conventional RT-PCR, demonstrating superior sensitivity for samples with low viral loads.
Why the study?
With the emergence of feline calicivirus-associated virulent systemic disease worldwide, a rapid, sensitive, and reproducible diagnostic assay is needed to inform prevention and control strategies.
Absolute Event Rate: 21% vs 5%
The established TaqMan RT-qPCR assay provides a highly sensitive and specific method for the rapid detection and quantification of Feline Calicivirus.
Feline calicivirus (FCV) is an infectious pathogen that causes disease in cats. With the current emergence of FCV-associated virulent systemic disease (FCV VSD) worldwide, the establishment of a rapid, sensitive, and reproducible diagnostic assay for its detection is important to inform prevention and control strategies. In this study, specific primers and TaqMan-FAM probes were designed based on the conserved regions of the FCV genome sequence, and a TaqMan reverse transcriptase quantitative real time PCR assay was established. This assay could specifically detected the FCV genome. The assay had a wide dynamic range, with linear detection in the range of 9.6×109 copies/μL to 9.6×100 copies/μL, with a limit of detection of 9.6×100 copies/μL, showing high sensitivity and repeatability. In addition, we used this assay to evaluated clinical samples (n=100) taken from cats from across China for the presence/absence of FCV genetic material For samples with low virus content, the positive detection rate of TaqMan reverse transcriptase quantitative real time PCR assay (RT-qPCR) was much higher than that of conventional reverse transcriptase PCR assay (cRT-PCR). And The qRT-PCR assay was used to detect the viral load of cat swabs within 17 days after FCV infection. From days 1-9, the oral and nasal swabs generally had higher viral loads than the anal swabs. While from days 10-17, the levels in the oral and nasal swabs being generally lower than those in the anal swabs. Overall, this FCV TaqMan RT-qPCR assay assay represents a rapid and accurate.
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zhao et al. (2021) studied Feline calicivirus (FCV) (n=100). TaqMan reverse transcriptase quantitative real-time PCR (qRT-PCR) vs. Conventional reverse transcriptase PCR (cRT-PCR) was evaluated on Positive detection rate of FCV in clinical samples. The TaqMan qRT-PCR assay detected feline calicivirus in 21% of clinical samples compared to only 5% using conventional RT-PCR, demonstrating superior sensitivity for samples with low viral loads.
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