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A Recombinase-Aided Amplification Assay for the Detection of Chlamydia felis

Jian Liu

Jian Liu

Shanghai Animal Disease Prevention and Control CenterChina
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Liu, Jian
, 
Weidong Qian

Weidong Qian

School of Food and Biological Engineering, Shaanxi University of Science and TechnologyXi’an, China
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Qian, Weidong
, 
Jian Wang

Jian Wang

Shanghai Animal Disease Prevention and Control CenterChina
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Wang, Jian
, 
Yilan Bai

Yilan Bai

Shanghai Animal Disease Prevention and Control CenterChina
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Bai, Yilan
, 
Yaping Gui

Yaping Gui

Shanghai Animal Disease Prevention and Control CenterChina
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Gui, Yaping
, 
Luming Xia

Luming Xia

Shanghai Animal Disease Prevention and Control CenterChina
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Xia, Luming
, 
Guohua Gong

Guohua Gong

Shanghai Animal Disease Prevention and Control CenterChina
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Gong, Guohua
, 
Feifei Ge

Feifei Ge

Shanghai Animal Disease Prevention and Control CenterChina
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Ge, Feifei
, 
Haixiao Shen

Haixiao Shen

Shanghai Animal Disease Prevention and Control CenterChina
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Shen, Haixiao
, 
Xiaojing Chang

Xiaojing Chang

Shanghai Animal Disease Prevention and Control CenterChina
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Chang, Xiaojing
 and 
Hongjin Zhao

Hongjin Zhao

Shanghai Animal Disease Prevention and Control CenterChina
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Zhao, Hongjin
  
Sep 20, 2023
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Article Category: Short Communication
Published Online: Sep 20, 2023
Page range: 339 - 343
Received: Mar 23, 2023
Accepted: Jun 30, 2023
DOI: https://doi.org/10.33073/pjm-2023-029
Keywords
© 2023 Jian Liu et al., published by Sciendo
This work is licensed under the Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International License.

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.
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.

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

Comparison of Chlamydia felis detection in 117 swabs examined by RAA and real-time PCR_

Real-time PCR Performance characteristics
Positive Negative Sensitivity Specificity
RAA Positive 20 0 95.24% 100%
Negative 1 96
Total 21 96
Language:
English
Publication timeframe:
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Journal Subjects:
Life Sciences, Microbiology and Virology
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