Zhuandı GONG, Xıaoyun SHEN, Haoqın LIANG, Jınjıng GENG, Suocheng WEI

TaqMan probe qRT-PCR detects bovine parvovirus and applies clinically

Öz The present study aimed to establish a TaqMan probe real-time PCR qRT-PCR for detecting Bovine parvovirus BPV , and to develop a novel diagnostic method of BPV. TaqMan probe and primers were designed and synthesized from the sequences of conserved 5?-untranslated regions 5'-UTR of strain Haden of BPV. Optimum reaction conditions were comparatively selected. The sensitivity, specificity, and reproducibility of TaqMan probe qRT-PCR were measured respectively. Feces specimens collected from diarrhea calves were detected using this assay. The results showed that optimized conditions for the probe qRT-PCR were 0.8 μL probes 18.0 μmol/L , 0.4 μL primers 10.0 μmol/L and annealing temperature of 43.2 °C for 40 s. The probe qRT-PCR could specially detect BPV, but not for other viruses JEV, CSF, RABV, BRV, BVDV, and bFMDV . The detection limit was 5.26 copies/μL. Both intraassay and interassay VCs were lower than 1.65%. Detection of 308 feces samples showed 36 positives for TaqMan probe qRT-PCR compared to 32 positives of universal PCR. Coincidence rate of the two methods was 88.89%. In conclusion, the established TaqMan probe qRT-PCR could specifically detect BPV. It had excellent sensitivity, specificity, and stability. The detection limit was 5.26 copies/μL. Its sensitivity was 10,000-fold over the conventional PCR. TaqMan probe qRT-PCR will help to enhance diagnosis and therapy efficacy of diarrhea calves.

Anahtar Kelimeler:

Bovine parvovirus, TaqMan probe, real-time PCR, sensitivity, specificity

Tam Metin

Kaynakça

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Kaynak Göster

Bibtex	`@ { tbtkveterinary724459, journal = {Turkish Journal of Veterinary and Animal Sciences}, issn = {1300-0128}, eissn = {1303-6181}, address = {}, publisher = {TÜBİTAK}, year = {2020}, volume = {44}, pages = {364 - 369}, doi = {10.3906/vet-1907-80}, title = {TaqMan probe qRT-PCR detects bovine parvovirus and applies clinically}, key = {cite}, author = {Gong, Zhuandı and Shen, Xıaoyun and Lıang, Haoqın and Geng, Jınjıng and Weı, Suocheng} }`
APA	Gong, Z , Shen, X , Lıang, H , Geng, J , Weı, S . (2020). TaqMan probe qRT-PCR detects bovine parvovirus and applies clinically . Turkish Journal of Veterinary and Animal Sciences , 44 (2) , 364-369 . DOI: 10.3906/vet-1907-80
MLA	Gong, Z , Shen, X , Lıang, H , Geng, J , Weı, S . "TaqMan probe qRT-PCR detects bovine parvovirus and applies clinically" . Turkish Journal of Veterinary and Animal Sciences 44 (2020 ): 364-369 <https://dergipark.org.tr/tr/pub/tbtkveterinary/issue/53838/724459>
Chicago	Gong, Z , Shen, X , Lıang, H , Geng, J , Weı, S . "TaqMan probe qRT-PCR detects bovine parvovirus and applies clinically". Turkish Journal of Veterinary and Animal Sciences 44 (2020 ): 364-369
RIS	TY - JOUR T1 - TaqMan probe qRT-PCR detects bovine parvovirus and applies clinically AU - Zhuandı Gong , Xıaoyun Shen , Haoqın Lıang , Jınjıng Geng , Suocheng Weı Y1 - 2020 PY - 2020 N1 - doi: 10.3906/vet-1907-80 DO - 10.3906/vet-1907-80 T2 - Turkish Journal of Veterinary and Animal Sciences JF - Journal JO - JOR SP - 364 EP - 369 VL - 44 IS - 2 SN - 1300-0128-1303-6181 M3 - doi: 10.3906/vet-1907-80 UR - https://doi.org/10.3906/vet-1907-80 Y2 - 2021 ER -
EndNote	%0 Turkish Journal of Veterinary and Animal Sciences TaqMan probe qRT-PCR detects bovine parvovirus and applies clinically %A Zhuandı Gong , Xıaoyun Shen , Haoqın Lıang , Jınjıng Geng , Suocheng Weı %T TaqMan probe qRT-PCR detects bovine parvovirus and applies clinically %D 2020 %J Turkish Journal of Veterinary and Animal Sciences %P 1300-0128-1303-6181 %V 44 %N 2 %R doi: 10.3906/vet-1907-80 %U 10.3906/vet-1907-80
ISNAD	Gong, Zhuandı , Shen, Xıaoyun , Lıang, Haoqın , Geng, Jınjıng , Weı, Suocheng . "TaqMan probe qRT-PCR detects bovine parvovirus and applies clinically". Turkish Journal of Veterinary and Animal Sciences 44 / 2 (Mart 2020): 364-369 . https://doi.org/10.3906/vet-1907-80
AMA	Gong Z , Shen X , Lıang H , Geng J , Weı S . TaqMan probe qRT-PCR detects bovine parvovirus and applies clinically. Turkish Journal of Veterinary and Animal Sciences. 2020; 44(2): 364-369.
Vancouver	Gong Z , Shen X , Lıang H , Geng J , Weı S . TaqMan probe qRT-PCR detects bovine parvovirus and applies clinically. Turkish Journal of Veterinary and Animal Sciences. 2020; 44(2): 364-369.
IEEE	Z. Gong , X. Shen , H. Lıang , J. Geng ve S. Weı , "TaqMan probe qRT-PCR detects bovine parvovirus and applies clinically", Turkish Journal of Veterinary and Animal Sciences, c. 44, sayı. 2, ss. 364-369, Mar. 2020, doi:10.3906/vet-1907-80