Journal: Journal of Virological Methods

Article Title: Characterization of a continuous feline mammary epithelial cell line susceptible to feline epitheliotropic viruses

doi: 10.1016/j.jviromet.2008.11.018

Figure Lengend Snippet: FMECs are susceptible to infection with feline epitheliotropic viruses. FMECs were inoculated with (A) FPV, (B) FCV, (C) FCoV, and (D) FHV then fixed and immunostained using virus-specific antibodies. Virus antigen is pseudocolored green. In panels A, C, and D, fJAM-A was immunostained and is pseudocolored red. Nuclei were stained with DAPI and pseudocolored magenta in panels B and C. Size bar in panel A = 23 μm, size bars in panels B–D = 50 μm.

Article Snippet: Cells were then fixed in 2% paraformaldehyde for 15–20 min at room temperature and then virus detected using the following antibodies: mouse anti-FCV (clone S1-8, Custom Monoclonals Intl., Sacramento, CA), mouse anti-parvovirus (to NS1 non structural protein, courtesy of C. Astell, University of British Columbia), mouse anti-canine coronavirus (FIPV-70, Custom Monoclonals Intl., Sacramento, CA) and mouse anti-feline herpesvirus (Herpes 7-7, Custom Monoclonals Intl., Sacramento, CA).

Techniques: Infection, Staining

Journal: Frontiers in Veterinary Science

Article Title: Development and Application of a Triplex TaqMan Quantitative Real-Time PCR Assay for Simultaneous Detection of Feline Calicivirus, Feline Parvovirus, and Feline Herpesvirus 1

doi: 10.3389/fvets.2021.792322

Figure Lengend Snippet: Specificity of the triplex assay. FPV, FCV, FHV-1, and other animal pathogens (including FIV, FeLV, FCoV, RV, and PRV) were selected and tested using the triplex assay. Only FCV, FHV-1, and FPV detected positive.

Article Snippet: FCV (attenuated vaccine strain F9), FHV-1 (attenuated vaccine strain G2620A), and FPV (attenuated vaccine strain MW-1) were purchased from Intervet International B.V. Wild strains of these three viruses were isolated and stored by our laboratory, and the virus titers were 10 8.0 TCID 50 /0.1 ml for FCV, 10 7.0 TCID 50 /0.1 ml for FPV, and 10 8.3 TCID 50 /0.1 ml for FHV-1.

Techniques:

Journal: Frontiers in Veterinary Science

Article Title: Development and Application of a Triplex TaqMan Quantitative Real-Time PCR Assay for Simultaneous Detection of Feline Calicivirus, Feline Parvovirus, and Feline Herpesvirus 1

doi: 10.3389/fvets.2021.792322

Figure Lengend Snippet: Sensitivity of the real-time PCR.

Article Snippet: FCV (attenuated vaccine strain F9), FHV-1 (attenuated vaccine strain G2620A), and FPV (attenuated vaccine strain MW-1) were purchased from Intervet International B.V. Wild strains of these three viruses were isolated and stored by our laboratory, and the virus titers were 10 8.0 TCID 50 /0.1 ml for FCV, 10 7.0 TCID 50 /0.1 ml for FPV, and 10 8.3 TCID 50 /0.1 ml for FHV-1.

Techniques: Real-time Polymerase Chain Reaction

Journal: Frontiers in Veterinary Science

Article Title: Development and Application of a Triplex TaqMan Quantitative Real-Time PCR Assay for Simultaneous Detection of Feline Calicivirus, Feline Parvovirus, and Feline Herpesvirus 1

doi: 10.3389/fvets.2021.792322

Figure Lengend Snippet: Detection limit of conventional PCR. Templates of pMD18T-FPV (A) , pMD18T-FCV-1 (B) , and pMD18T-FHV-1 (C) were diluted by 10 times gradient to a dilution factor that could not be detected by conventional PCR. The detection limit was 5 × 10 3 copies/assay for FPV and 5 × 10 2 copies/assay for FCV and FHV-1. Template amount for curves 2–8 lanes was 5 × 10 7 -5 × 10 1 copies/assay. M, DL600 marker; NC, negative control.

Article Snippet: FCV (attenuated vaccine strain F9), FHV-1 (attenuated vaccine strain G2620A), and FPV (attenuated vaccine strain MW-1) were purchased from Intervet International B.V. Wild strains of these three viruses were isolated and stored by our laboratory, and the virus titers were 10 8.0 TCID 50 /0.1 ml for FCV, 10 7.0 TCID 50 /0.1 ml for FPV, and 10 8.3 TCID 50 /0.1 ml for FHV-1.

Techniques: Marker, Negative Control

Journal: Frontiers in Veterinary Science

Article Title: Development and Application of a Triplex TaqMan Quantitative Real-Time PCR Assay for Simultaneous Detection of Feline Calicivirus, Feline Parvovirus, and Feline Herpesvirus 1

doi: 10.3389/fvets.2021.792322

Figure Lengend Snippet: Amplification and standard curves of the triplex assay. Serially diluted plasmids were mixed at equal concentration (from 5 × 10 7 copies/assay to 5 × 10 1 copies/assay) and used as template for RT-PCR. Amplification curves of the triplex assay for the detection of FPV (A) , FCV (C) , and FHV-1 (E) . The standard curves of FPV (B) , FCV (D) , and FHV-1 (F) were generated by plotting the Ct values (Y-axis) against the logarithm of copy numbers of plasmids (X-axis).

Article Snippet: FCV (attenuated vaccine strain F9), FHV-1 (attenuated vaccine strain G2620A), and FPV (attenuated vaccine strain MW-1) were purchased from Intervet International B.V. Wild strains of these three viruses were isolated and stored by our laboratory, and the virus titers were 10 8.0 TCID 50 /0.1 ml for FCV, 10 7.0 TCID 50 /0.1 ml for FPV, and 10 8.3 TCID 50 /0.1 ml for FHV-1.

Techniques: Amplification, Concentration Assay, Reverse Transcription Polymerase Chain Reaction, Generated

Journal: Frontiers in Veterinary Science

Article Title: Development and Application of a Triplex TaqMan Quantitative Real-Time PCR Assay for Simultaneous Detection of Feline Calicivirus, Feline Parvovirus, and Feline Herpesvirus 1

doi: 10.3389/fvets.2021.792322

Figure Lengend Snippet: Intra- and inter-assay reproducibility of the triplex assay.

Article Snippet: FCV (attenuated vaccine strain F9), FHV-1 (attenuated vaccine strain G2620A), and FPV (attenuated vaccine strain MW-1) were purchased from Intervet International B.V. Wild strains of these three viruses were isolated and stored by our laboratory, and the virus titers were 10 8.0 TCID 50 /0.1 ml for FCV, 10 7.0 TCID 50 /0.1 ml for FPV, and 10 8.3 TCID 50 /0.1 ml for FHV-1.

Techniques: Inter Assay, Intra Assay

Journal: Frontiers in Veterinary Science

Article Title: Development and Application of a Triplex TaqMan Quantitative Real-Time PCR Assay for Simultaneous Detection of Feline Calicivirus, Feline Parvovirus, and Feline Herpesvirus 1

doi: 10.3389/fvets.2021.792322

Figure Lengend Snippet: Detection of the co-infection models by triplex real-time PCR.

Article Snippet: FCV (attenuated vaccine strain F9), FHV-1 (attenuated vaccine strain G2620A), and FPV (attenuated vaccine strain MW-1) were purchased from Intervet International B.V. Wild strains of these three viruses were isolated and stored by our laboratory, and the virus titers were 10 8.0 TCID 50 /0.1 ml for FCV, 10 7.0 TCID 50 /0.1 ml for FPV, and 10 8.3 TCID 50 /0.1 ml for FHV-1.

Techniques: Virus, Real-time Polymerase Chain Reaction

Journal: Frontiers in Veterinary Science

Article Title: Development and Application of a Triplex TaqMan Quantitative Real-Time PCR Assay for Simultaneous Detection of Feline Calicivirus, Feline Parvovirus, and Feline Herpesvirus 1

doi: 10.3389/fvets.2021.792322

Figure Lengend Snippet: Clinical samples detected by triplex assay and commercial kits.

Article Snippet: FCV (attenuated vaccine strain F9), FHV-1 (attenuated vaccine strain G2620A), and FPV (attenuated vaccine strain MW-1) were purchased from Intervet International B.V. Wild strains of these three viruses were isolated and stored by our laboratory, and the virus titers were 10 8.0 TCID 50 /0.1 ml for FCV, 10 7.0 TCID 50 /0.1 ml for FPV, and 10 8.3 TCID 50 /0.1 ml for FHV-1.

Techniques: