Establishment and preliminary application of dual fluorescent quantitative PCR for detection of RV and CAV2
- VernacularTitle:狂犬病病毒和犬2型腺病毒双重荧光定量PCR检测方法的建立及应用
- Author:
Jian LIU
1
;
Yaping GUI
1
;
Yilan BAI
1
;
Luming XIA
1
;
Xiaoying ZHU
1
;
Xianchao YANG
1
;
Tiangusheng TAO
1
;
Congsheng TANG
1
;
Yujie ZHANG
1
;
Jian WANG
1
;
Hongjin ZHAO
1
Author Information
- Publication Type:Journal Article
- Keywords: Rabies virus; Canine adenovirus type 2; Dual fluorescent quantitative PCR; CAV2-ΔE3-CGS
- From: Journal of Public Health and Preventive Medicine 2023;34(3):33-37
- CountryChina
- Language:Chinese
- Abstract: Objective To investigate the shedding of CAV2-ΔE3-CGS after immunization and the background of canine adenovirus (CAV) infection, and to establish a dual fluorescent quantitative PCR detection method for rabies virus (RV) and canine adenovirus type 2 (CAV2). Methods A dual fluorescent quantitative PCR detection method was established by designing specific primers and probes for E1 gene of CAV and G gene of RV for the detection of CAV2-ΔE3-CGS. Oral swabs, anal swabs and environmental samples of stray dogs from experimental animal farm and dog detention center were tested. Results The standard curves generated by this method were Y=-3.351 × logX + 44.895, R2 = .999 and Y=-3.413 × logX + 45.192, R2=0.996, respectively. The linear relationships were good, and the minimum detection limits were both 102 copies/μL. CAV2-ΔE3-CGS was not detected in experimental animal farm. CAV was detected in dog detention center, and the positive rates were 5.88% (5/85) in oral swabs, 8.24% (7/85) in anal swabs, and 4% (1/25) in environmental samples. Conclusion The dual fluorescent quantitative PCR method can be used for the detection of CAV2-ΔE3-CGS after immunization and the investigation of CAV infection. The present study has shown that no CAV2-ΔE3-CGS has been detected after immunization and CAV infection rate of stay dogs is low in Shanghai. CAV2-ΔE3-CGS oral immunization meets requirement and is applicable.
