Development of a time-resolved fluorescence immunoassay kit for detecting canine coronavirus and parvovirus through double labeling
Abstract Objective Canine enteric coronavirus (CCV) and canine parvovirus type 2 (CPV-2) are the main pathogens responsible for acute gastroenteritis in dogs, and both single and mixed infections are common. This study aimed to establish a double-labeling time-resolved fluorescence immunoassay (TRFI...
| Main Authors: | , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
BMC
2024-03-01
|
| Series: | Virology Journal |
| Subjects: | |
| Online Access: | https://doi.org/10.1186/s12985-024-02302-4 |
| _version_ | 1797259482536869888 |
|---|---|
| author | Laiqing Li Cuicui Chen Huankun Liang Wenqi Dong V. N. Leontiev Igor Vitalievich Voytov |
| author_facet | Laiqing Li Cuicui Chen Huankun Liang Wenqi Dong V. N. Leontiev Igor Vitalievich Voytov |
| author_sort | Laiqing Li |
| collection | DOAJ |
| description | Abstract Objective Canine enteric coronavirus (CCV) and canine parvovirus type 2 (CPV-2) are the main pathogens responsible for acute gastroenteritis in dogs, and both single and mixed infections are common. This study aimed to establish a double-labeling time-resolved fluorescence immunoassay (TRFIA) to test and distinguish CCV and CPV-2 diseases. Methods A sandwich double-labeling TRFIA method was established and optimized using europium(III) (Eu3+)/samarium(III) (Sm3+) chelates. CCV/CPV-2 antigens were first captured by the immobilized antibodies. Then, combined with Eu3+/Sm3+-labeled paired antibodies, the Eu3+/Sm3+ fluorescence values were detected after dissociation to calculate the CCV/CPV-2 ratios. The performance, clinical performance and methodology used for laboratory (sensitivity, specificity, accuracy and stability) testing were evaluated. Results A double-label TRFIA for CCV and CPV-2 detection was optimized and established. The sensitivity of this TRFIA kit was 0.51 ng/mL for CCV and 0.80 ng/mL for CPV-2, with high specificity for CCV and CPV-2. All the accuracy data were less than 10%, and the recovery ranged from 101.21 to 110.28%. The kits can be temporarily stored for 20 days at 4 °C and can be stored for 12 months at temperatures less than − 20 °C. Based on a methodology comparison of 137 clinically suspected patients, there was no statistically significant difference between the TRFIA kit and the PCR method. Additionally, for CCV detection, the clinical sensitivity was 95.74%, and the clinical specificity was 93.33%. For CPV-2 detection, the clinical sensitivity was 92.86%, and the clinical specificity was 96.97%. Conclusion In this study, a double-label TRFIA kit was prepared for CCV and CPV-2 detection with high laboratory sensitivity, specificity, accuracy, stability, clinical sensitivity and specificity. This kit provides a new option for screening/distinguishing between CCV and CPV-2 and may help improve strategies to prevent and control animal infectious diseases in the future. |
| first_indexed | 2024-04-24T23:10:08Z |
| format | Article |
| id | doaj.art-2d005f4a09ef462885c6e2a2aa5e8932 |
| institution | Directory Open Access Journal |
| issn | 1743-422X |
| language | English |
| last_indexed | 2024-04-24T23:10:08Z |
| publishDate | 2024-03-01 |
| publisher | BMC |
| record_format | Article |
| series | Virology Journal |
| spelling | doaj.art-2d005f4a09ef462885c6e2a2aa5e89322024-03-17T12:14:26ZengBMCVirology Journal1743-422X2024-03-012111810.1186/s12985-024-02302-4Development of a time-resolved fluorescence immunoassay kit for detecting canine coronavirus and parvovirus through double labelingLaiqing Li0Cuicui Chen1Huankun Liang2Wenqi Dong3V. N. Leontiev4Igor Vitalievich Voytov5Belarusian State Technological UniversityGuangzhou Youdi Bio-technology Co., LtdGuangzhou Youdi Bio-technology Co., LtdGuangzhou Zhenda Biopharmaceutical Technology Co., LtdBelarusian State Technological UniversityBelarusian State Technological UniversityAbstract Objective Canine enteric coronavirus (CCV) and canine parvovirus type 2 (CPV-2) are the main pathogens responsible for acute gastroenteritis in dogs, and both single and mixed infections are common. This study aimed to establish a double-labeling time-resolved fluorescence immunoassay (TRFIA) to test and distinguish CCV and CPV-2 diseases. Methods A sandwich double-labeling TRFIA method was established and optimized using europium(III) (Eu3+)/samarium(III) (Sm3+) chelates. CCV/CPV-2 antigens were first captured by the immobilized antibodies. Then, combined with Eu3+/Sm3+-labeled paired antibodies, the Eu3+/Sm3+ fluorescence values were detected after dissociation to calculate the CCV/CPV-2 ratios. The performance, clinical performance and methodology used for laboratory (sensitivity, specificity, accuracy and stability) testing were evaluated. Results A double-label TRFIA for CCV and CPV-2 detection was optimized and established. The sensitivity of this TRFIA kit was 0.51 ng/mL for CCV and 0.80 ng/mL for CPV-2, with high specificity for CCV and CPV-2. All the accuracy data were less than 10%, and the recovery ranged from 101.21 to 110.28%. The kits can be temporarily stored for 20 days at 4 °C and can be stored for 12 months at temperatures less than − 20 °C. Based on a methodology comparison of 137 clinically suspected patients, there was no statistically significant difference between the TRFIA kit and the PCR method. Additionally, for CCV detection, the clinical sensitivity was 95.74%, and the clinical specificity was 93.33%. For CPV-2 detection, the clinical sensitivity was 92.86%, and the clinical specificity was 96.97%. Conclusion In this study, a double-label TRFIA kit was prepared for CCV and CPV-2 detection with high laboratory sensitivity, specificity, accuracy, stability, clinical sensitivity and specificity. This kit provides a new option for screening/distinguishing between CCV and CPV-2 and may help improve strategies to prevent and control animal infectious diseases in the future.https://doi.org/10.1186/s12985-024-02302-4CoronavirusParvovirusCanineTime-resolved fluorescence immunoassayDouble labelingKit |
| spellingShingle | Laiqing Li Cuicui Chen Huankun Liang Wenqi Dong V. N. Leontiev Igor Vitalievich Voytov Development of a time-resolved fluorescence immunoassay kit for detecting canine coronavirus and parvovirus through double labeling Virology Journal Coronavirus Parvovirus Canine Time-resolved fluorescence immunoassay Double labeling Kit |
| title | Development of a time-resolved fluorescence immunoassay kit for detecting canine coronavirus and parvovirus through double labeling |
| title_full | Development of a time-resolved fluorescence immunoassay kit for detecting canine coronavirus and parvovirus through double labeling |
| title_fullStr | Development of a time-resolved fluorescence immunoassay kit for detecting canine coronavirus and parvovirus through double labeling |
| title_full_unstemmed | Development of a time-resolved fluorescence immunoassay kit for detecting canine coronavirus and parvovirus through double labeling |
| title_short | Development of a time-resolved fluorescence immunoassay kit for detecting canine coronavirus and parvovirus through double labeling |
| title_sort | development of a time resolved fluorescence immunoassay kit for detecting canine coronavirus and parvovirus through double labeling |
| topic | Coronavirus Parvovirus Canine Time-resolved fluorescence immunoassay Double labeling Kit |
| url | https://doi.org/10.1186/s12985-024-02302-4 |
| work_keys_str_mv | AT laiqingli developmentofatimeresolvedfluorescenceimmunoassaykitfordetectingcaninecoronavirusandparvovirusthroughdoublelabeling AT cuicuichen developmentofatimeresolvedfluorescenceimmunoassaykitfordetectingcaninecoronavirusandparvovirusthroughdoublelabeling AT huankunliang developmentofatimeresolvedfluorescenceimmunoassaykitfordetectingcaninecoronavirusandparvovirusthroughdoublelabeling AT wenqidong developmentofatimeresolvedfluorescenceimmunoassaykitfordetectingcaninecoronavirusandparvovirusthroughdoublelabeling AT vnleontiev developmentofatimeresolvedfluorescenceimmunoassaykitfordetectingcaninecoronavirusandparvovirusthroughdoublelabeling AT igorvitalievichvoytov developmentofatimeresolvedfluorescenceimmunoassaykitfordetectingcaninecoronavirusandparvovirusthroughdoublelabeling |