Two Multiplex PCR Methods for Detecting Several Pathogens Associated with Feline Respiratory and Intestinal Tracts
Abstract
:Simple Summary
Abstract
1. Introduction
2. Materials and Methods
2.1. Viruses and Bacterial Strains
2.2. Primer Design
2.3. Nucleic Acid Extraction and Reverse Transcription and Construction of Standard Plasmids
2.4. Establishment of the mPCR Methods
2.5. Specificity of the mPCR Methods
2.6. Sensitivity of mPCR Methods
2.7. Examination of Clinical Samples
3. Results
3.1. Optimum Annealing Temperature for the mPCR Methods
3.2. Specificity of mPCR Methods
3.3. Sensitivity of the mPCR Methods
3.4. Examination of Clinical Samples
4. Discussion
5. Conclusions
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Acknowledgments
Conflicts of Interest
References
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Pathogen | Primer Name | Primer Sequence (5′-3′) | Target Gene | PCR Products |
---|---|---|---|---|
FCV | FCV-F | AACCTGCGCTAACGTGCT | VP1 | 1940 bp |
FCV-R | TGWATTCCCATGTAGGAGGC | |||
FHV-1 | FHV-1-F | TACTTCAAGCCTTACGACCACG | gD | 1014 bp |
FHV-1-R | GATCGAGACCTCTTTACCCTCA | |||
FeLV | FeLV-F | TGGCACAGGTAAAGCAAGTT | gPr80 | 608 bp |
FeLV-R | CGTGTCCACCAARAARGTCA | |||
C.felis | C.felis-F | TTGTCGGATTGATTGGTCTT | ompA | 500 bp |
C.felis-R | AGTTGGGTTCCAGGTTGTTA | |||
IAV | IAV-F | CAGAGACTKGAARRTGTCTTTGC | M | 155 bp |
IAV-R | CTACGCTGCAGTCCTCGCTC | |||
FCoV | FCoV-F | GATGGAGTMTTCTGGGTTGC | N | 726 bp |
FCoV-R | TTCCTCAGGCTTCTTATCAG | |||
FeAstV | FeAstV-F | GAAATGGACTGGACACGTTATGA | ORF1ab | 418 bp |
FeAstV-R | GGCTTGACCCACATGCCGAA | |||
FPV | FPV-F | AAGACGTGCAAGCGAGTCC | VP2 | 337 bp |
FPV-R | GAGCGAAGATAAGCAGCGTAA | |||
FeKoV | FeKoV-F | CCTCTTTYCTTCGGGACTTTTA | polyprotein | 198 bp |
FeKoV-R | ACCACATCACTGAYTGTTCGTA |
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Xiao, X.; Hao, X.; Chen, B.; Zhou, P.; Li, S. Two Multiplex PCR Methods for Detecting Several Pathogens Associated with Feline Respiratory and Intestinal Tracts. Vet. Sci. 2023, 10, 14. https://doi.org/10.3390/vetsci10010014
Xiao X, Hao X, Chen B, Zhou P, Li S. Two Multiplex PCR Methods for Detecting Several Pathogens Associated with Feline Respiratory and Intestinal Tracts. Veterinary Sciences. 2023; 10(1):14. https://doi.org/10.3390/vetsci10010014
Chicago/Turabian StyleXiao, Xiangyu, Xiangqi Hao, Bo Chen, Pei Zhou, and Shoujun Li. 2023. "Two Multiplex PCR Methods for Detecting Several Pathogens Associated with Feline Respiratory and Intestinal Tracts" Veterinary Sciences 10, no. 1: 14. https://doi.org/10.3390/vetsci10010014