Table I
Sequences of primers and probes in the RAA assay for Chlamydia felis.*
| Primers/Probes | Sequence (5′–3′) | Genomic position† |
|---|---|---|
| C. felis F | GAAAGCAAGGGGAGCAAACAGGATT | 769–793 |
| C. felis R | TCAGGCGGCATACTTAACGCGTTAG | 891–867 |
| C. felis probe | CGATGCATACTTGATGTGGATAGTCTCAACCC[FAM-dT] A(THF)[BHQ1-dT] CCGTGTCGTAGCTAAC(C3-Spacer) | 821–871 |
Fig. 1.
A) Specificity evaluation of the recombinase-aided amplification (RAA) assay for the detection of Chlamydia felis using genomic DNA from C. felis_pUC57 plasmid, feline calicivirus-SH202101 (FCV), feline herpesvirus-1/Cat/Shanghai/01/2014 (FHV-1), feline infectious peritonitis virus VR-990 (FIPV), feline parvovirus/Shanghai/01/2021 (FPV), Mycoplasma felis ATCC® 23391™ (M. felis), and Bordetella bronchiseptica ATCC® 4617™ (B. bronchiseptica), as the templates of RAA, respectively.
B) Examination of the RAA assay's sensitivity for detecting Chlamydia felis using a serial dilution of the C. felis_pUC57 plasmid as the template. The lower limit of detection was 10.6 copies/reaction plasmid DNA. The higher the template concentration, the greater the fluorescence value and the earlier the peak time. NC, negative control group. All the experiments were performed at least three times.