Development of a Synthetic VP1 Protein Peptide-Based ELISA to Detect Antibodies Against Porcine Bocavirus Group 3

Porcine bocavirus (PBoV), classified within the genus Bocaparvovirus, has been reported worldwide. PBoV has been divided into group 1, group 2, and group 3. PBoV group 3 (G3) viruses are the most prevalent in China. Currently, effective serological methods for the detection of antibodies against PBo...

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Main Authors: Chao Gong, Hui He, Yuguang Fu, Baoyu Li, Bin Yang, Jianlong Li, Xiaodong He, Juncheng Han, Yi Zhang, Guangliang Liu, Qingyong Guo
Format: Article
Language:English
Published: MDPI AG 2024-12-01
Series:Viruses
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Online Access:https://www.mdpi.com/1999-4915/16/12/1946
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author Chao Gong
Hui He
Yuguang Fu
Baoyu Li
Bin Yang
Jianlong Li
Xiaodong He
Juncheng Han
Yi Zhang
Guangliang Liu
Qingyong Guo
author_facet Chao Gong
Hui He
Yuguang Fu
Baoyu Li
Bin Yang
Jianlong Li
Xiaodong He
Juncheng Han
Yi Zhang
Guangliang Liu
Qingyong Guo
author_sort Chao Gong
collection DOAJ
description Porcine bocavirus (PBoV), classified within the genus Bocaparvovirus, has been reported worldwide. PBoV has been divided into group 1, group 2, and group 3. PBoV group 3 (G3) viruses are the most prevalent in China. Currently, effective serological methods for the detection of antibodies against PBoV G3 are limited. In this study, we developed an indirect ELISA using a synthetic VP1 peptide designed on the basis of the conserved region of the PBoV VP1 protein as a coating antigen. Through matrix titration, the optimal coating concentration of the VP1 peptide (0.5 μg/mL), serum dilution (1:200), and working concentration of the secondary antibody (1:50,000) were determined. The cutoff value of this developed ELISA was set as 0.4239. Further investigations revealed that this developed ELISA had no cross-reactivity with positive serum antibodies against FMDV-O, FMDV-A, PRV, ASFV, SF, PCV2, PEDV, and TGEV. The detection limit of the method was a 1:1600 dilution of standard positive serum against PBoV G3. The coefficients of variation for both the intra- and interassay data were lower than 10%. A total of 1373 serum samples collected from 12 provinces in China between 2022 and 2023 were subjected to indirect ELISA. The results showed that 47.56% of the samples were PBoV G3 positive. These results reveal that peptide-based ELISA is a reliable and cost-effective method for detecting PBoV G3 antibodies. It also facilitates the investigation of the prevalence and distribution of PBoV G3.
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spelling doaj-art-8091ccab4be44e209f15d969ee0f3ff62025-08-20T02:43:54ZengMDPI AGViruses1999-49152024-12-011612194610.3390/v16121946Development of a Synthetic VP1 Protein Peptide-Based ELISA to Detect Antibodies Against Porcine Bocavirus Group 3Chao Gong0Hui He1Yuguang Fu2Baoyu Li3Bin Yang4Jianlong Li5Xiaodong He6Juncheng Han7Yi Zhang8Guangliang Liu9Qingyong Guo10Xinjiang Key Laboratory of New Drug Study and Creation for Herbivorous Animals (XJ-KLNDSCHA), College of Veterinary Medicine, Xinjiang Agricultural University, Urumqi 830052, ChinaXinjiang Key Laboratory of New Drug Study and Creation for Herbivorous Animals (XJ-KLNDSCHA), College of Veterinary Medicine, Xinjiang Agricultural University, Urumqi 830052, ChinaState Key Laboratory for Animal Disease Control and Prevention, College of Veterinary Medicine, Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Lanzhou University, Lanzhou 730000, ChinaState Key Laboratory for Animal Disease Control and Prevention, College of Veterinary Medicine, Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Lanzhou University, Lanzhou 730000, ChinaState Key Laboratory for Animal Disease Control and Prevention, College of Veterinary Medicine, Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Lanzhou University, Lanzhou 730000, ChinaXinjiang Key Laboratory of New Drug Study and Creation for Herbivorous Animals (XJ-KLNDSCHA), College of Veterinary Medicine, Xinjiang Agricultural University, Urumqi 830052, ChinaXinjiang Key Laboratory of New Drug Study and Creation for Herbivorous Animals (XJ-KLNDSCHA), College of Veterinary Medicine, Xinjiang Agricultural University, Urumqi 830052, ChinaXinjiang Key Laboratory of New Drug Study and Creation for Herbivorous Animals (XJ-KLNDSCHA), College of Veterinary Medicine, Xinjiang Agricultural University, Urumqi 830052, ChinaXinjiang Key Laboratory of New Drug Study and Creation for Herbivorous Animals (XJ-KLNDSCHA), College of Veterinary Medicine, Xinjiang Agricultural University, Urumqi 830052, ChinaXinjiang Key Laboratory of New Drug Study and Creation for Herbivorous Animals (XJ-KLNDSCHA), College of Veterinary Medicine, Xinjiang Agricultural University, Urumqi 830052, ChinaXinjiang Key Laboratory of New Drug Study and Creation for Herbivorous Animals (XJ-KLNDSCHA), College of Veterinary Medicine, Xinjiang Agricultural University, Urumqi 830052, ChinaPorcine bocavirus (PBoV), classified within the genus Bocaparvovirus, has been reported worldwide. PBoV has been divided into group 1, group 2, and group 3. PBoV group 3 (G3) viruses are the most prevalent in China. Currently, effective serological methods for the detection of antibodies against PBoV G3 are limited. In this study, we developed an indirect ELISA using a synthetic VP1 peptide designed on the basis of the conserved region of the PBoV VP1 protein as a coating antigen. Through matrix titration, the optimal coating concentration of the VP1 peptide (0.5 μg/mL), serum dilution (1:200), and working concentration of the secondary antibody (1:50,000) were determined. The cutoff value of this developed ELISA was set as 0.4239. Further investigations revealed that this developed ELISA had no cross-reactivity with positive serum antibodies against FMDV-O, FMDV-A, PRV, ASFV, SF, PCV2, PEDV, and TGEV. The detection limit of the method was a 1:1600 dilution of standard positive serum against PBoV G3. The coefficients of variation for both the intra- and interassay data were lower than 10%. A total of 1373 serum samples collected from 12 provinces in China between 2022 and 2023 were subjected to indirect ELISA. The results showed that 47.56% of the samples were PBoV G3 positive. These results reveal that peptide-based ELISA is a reliable and cost-effective method for detecting PBoV G3 antibodies. It also facilitates the investigation of the prevalence and distribution of PBoV G3.https://www.mdpi.com/1999-4915/16/12/1946peptide-based ELISAVP1porcine bocavirus group 3
spellingShingle Chao Gong
Hui He
Yuguang Fu
Baoyu Li
Bin Yang
Jianlong Li
Xiaodong He
Juncheng Han
Yi Zhang
Guangliang Liu
Qingyong Guo
Development of a Synthetic VP1 Protein Peptide-Based ELISA to Detect Antibodies Against Porcine Bocavirus Group 3
Viruses
peptide-based ELISA
VP1
porcine bocavirus group 3
title Development of a Synthetic VP1 Protein Peptide-Based ELISA to Detect Antibodies Against Porcine Bocavirus Group 3
title_full Development of a Synthetic VP1 Protein Peptide-Based ELISA to Detect Antibodies Against Porcine Bocavirus Group 3
title_fullStr Development of a Synthetic VP1 Protein Peptide-Based ELISA to Detect Antibodies Against Porcine Bocavirus Group 3
title_full_unstemmed Development of a Synthetic VP1 Protein Peptide-Based ELISA to Detect Antibodies Against Porcine Bocavirus Group 3
title_short Development of a Synthetic VP1 Protein Peptide-Based ELISA to Detect Antibodies Against Porcine Bocavirus Group 3
title_sort development of a synthetic vp1 protein peptide based elisa to detect antibodies against porcine bocavirus group 3
topic peptide-based ELISA
VP1
porcine bocavirus group 3
url https://www.mdpi.com/1999-4915/16/12/1946
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