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Simultaneous detection of feline parvovirus and feline bocavirus using SYBR Green I-based duplex real-time polymerase chain reaction

Since both feline parvovirus (FPV) and feline bocavirus (FBoV) can cause diarrhea in cats, it is difficult to distinguish them clinically. This study aimed to develop a SYBR Green I-based duplex real-time polymerase chain reaction (PCR) assay for distinguishing FPV and FBoV-1 on the basis of the mel...

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Autores principales: Wang, Yong, Pan, Yang, Wu, Junhuang, Tong, Xinxin, Sun, Jianfei, Xu, Fazhi, Cheng, Bangzhao, Li, Yongdong
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Springer International Publishing 2021
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8343365/
https://www.ncbi.nlm.nih.gov/pubmed/34377624
http://dx.doi.org/10.1007/s13205-021-02947-w
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author Wang, Yong
Pan, Yang
Wu, Junhuang
Tong, Xinxin
Sun, Jianfei
Xu, Fazhi
Cheng, Bangzhao
Li, Yongdong
author_facet Wang, Yong
Pan, Yang
Wu, Junhuang
Tong, Xinxin
Sun, Jianfei
Xu, Fazhi
Cheng, Bangzhao
Li, Yongdong
author_sort Wang, Yong
collection PubMed
description Since both feline parvovirus (FPV) and feline bocavirus (FBoV) can cause diarrhea in cats, it is difficult to distinguish them clinically. This study aimed to develop a SYBR Green I-based duplex real-time polymerase chain reaction (PCR) assay for distinguishing FPV and FBoV-1 on the basis of the melting temperature of the PCR product. A total of 132 fecal samples from different domestic and feral cats were collected, and the results of SYBR Green I-based duplex real-time PCR assay were compared with those of the traditional PCR assay for a comprehensive evaluation. The melting temperatures were found to be 86 °C and 77.5 °C for FBoV-1 and FPV, respectively, and no specific melting peaks for other non-targeted feline viruses were observed. The data obtained from this assay had a good linear relationship; the detection limits of FPV and FBoV-1 were 2.907 × 10(1) copies/μL and 3.836 × 10(1) copies/μL, respectively. In addition, the experiment exhibited high reproducibility. The positive detection rates of the SYBR Green I-based duplex real-time PCR assay for FPV and FBoV-1 were 16.67% (22/132) and 6.82% (9/132), respectively, and the positive detection rate for co-infection with FPV and FBoV-1 was 3.03% (4/132). This result was much more sensitive than that of the traditional PCR method. Thus, the developed SYBR Green I-based assay is a sensitive, rapid, specific, and reliable method for the clinical diagnosis of FPV and FBoV-1 and can provide technical support for the simultaneous detection of co-infection with these viruses in the future. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1007/s13205-021-02947-w.
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spelling pubmed-83433652021-08-06 Simultaneous detection of feline parvovirus and feline bocavirus using SYBR Green I-based duplex real-time polymerase chain reaction Wang, Yong Pan, Yang Wu, Junhuang Tong, Xinxin Sun, Jianfei Xu, Fazhi Cheng, Bangzhao Li, Yongdong 3 Biotech Original Article Since both feline parvovirus (FPV) and feline bocavirus (FBoV) can cause diarrhea in cats, it is difficult to distinguish them clinically. This study aimed to develop a SYBR Green I-based duplex real-time polymerase chain reaction (PCR) assay for distinguishing FPV and FBoV-1 on the basis of the melting temperature of the PCR product. A total of 132 fecal samples from different domestic and feral cats were collected, and the results of SYBR Green I-based duplex real-time PCR assay were compared with those of the traditional PCR assay for a comprehensive evaluation. The melting temperatures were found to be 86 °C and 77.5 °C for FBoV-1 and FPV, respectively, and no specific melting peaks for other non-targeted feline viruses were observed. The data obtained from this assay had a good linear relationship; the detection limits of FPV and FBoV-1 were 2.907 × 10(1) copies/μL and 3.836 × 10(1) copies/μL, respectively. In addition, the experiment exhibited high reproducibility. The positive detection rates of the SYBR Green I-based duplex real-time PCR assay for FPV and FBoV-1 were 16.67% (22/132) and 6.82% (9/132), respectively, and the positive detection rate for co-infection with FPV and FBoV-1 was 3.03% (4/132). This result was much more sensitive than that of the traditional PCR method. Thus, the developed SYBR Green I-based assay is a sensitive, rapid, specific, and reliable method for the clinical diagnosis of FPV and FBoV-1 and can provide technical support for the simultaneous detection of co-infection with these viruses in the future. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1007/s13205-021-02947-w. Springer International Publishing 2021-08-06 2021-09 /pmc/articles/PMC8343365/ /pubmed/34377624 http://dx.doi.org/10.1007/s13205-021-02947-w Text en © King Abdulaziz City for Science and Technology 2021
spellingShingle Original Article
Wang, Yong
Pan, Yang
Wu, Junhuang
Tong, Xinxin
Sun, Jianfei
Xu, Fazhi
Cheng, Bangzhao
Li, Yongdong
Simultaneous detection of feline parvovirus and feline bocavirus using SYBR Green I-based duplex real-time polymerase chain reaction
title Simultaneous detection of feline parvovirus and feline bocavirus using SYBR Green I-based duplex real-time polymerase chain reaction
title_full Simultaneous detection of feline parvovirus and feline bocavirus using SYBR Green I-based duplex real-time polymerase chain reaction
title_fullStr Simultaneous detection of feline parvovirus and feline bocavirus using SYBR Green I-based duplex real-time polymerase chain reaction
title_full_unstemmed Simultaneous detection of feline parvovirus and feline bocavirus using SYBR Green I-based duplex real-time polymerase chain reaction
title_short Simultaneous detection of feline parvovirus and feline bocavirus using SYBR Green I-based duplex real-time polymerase chain reaction
title_sort simultaneous detection of feline parvovirus and feline bocavirus using sybr green i-based duplex real-time polymerase chain reaction
topic Original Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8343365/
https://www.ncbi.nlm.nih.gov/pubmed/34377624
http://dx.doi.org/10.1007/s13205-021-02947-w
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