Development of high-concentration labeled colloidal gold immunochromatographic test strips for detecting african swine fever virus p30 protein antibodies

African Swine Fever (ASF), caused by the African swine fever virus (ASFV), has inflicted significant economic losses on the pig industry in China. The key to mitigating its impact lies in accurate screening and strict biosecurity measures. In this regard, the development of colloidal gold immunochro...

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Main Authors: Huai-cheng Liu, Rong-chao Liu, Mei-rong Hu, Ao-bing Yang, Ren-hu Wu, Yan Chen, Jin Zhang, Ji-shan Bai, Sheng-bo Wu, Jian-peng Chen, Yun-feng Long, Yan Jiang, Bin Zhou
Format: Article
Language:English
Published: Elsevier 2024-02-01
Series:Heliyon
Subjects:
Online Access:http://www.sciencedirect.com/science/article/pii/S2405844024012453
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author Huai-cheng Liu
Rong-chao Liu
Mei-rong Hu
Ao-bing Yang
Ren-hu Wu
Yan Chen
Jin Zhang
Ji-shan Bai
Sheng-bo Wu
Jian-peng Chen
Yun-feng Long
Yan Jiang
Bin Zhou
author_facet Huai-cheng Liu
Rong-chao Liu
Mei-rong Hu
Ao-bing Yang
Ren-hu Wu
Yan Chen
Jin Zhang
Ji-shan Bai
Sheng-bo Wu
Jian-peng Chen
Yun-feng Long
Yan Jiang
Bin Zhou
author_sort Huai-cheng Liu
collection DOAJ
description African Swine Fever (ASF), caused by the African swine fever virus (ASFV), has inflicted significant economic losses on the pig industry in China. The key to mitigating its impact lies in accurate screening and strict biosecurity measures. In this regard, the development of colloidal gold immunochromatographic test strips (CGITS) has proven to be an effective method for detecting ASFV antibodies. These test strips are based on the ASFV p30 recombinant protein and corresponding monoclonal antibodies. The design of the test strip incorporates a high-concentration colloidal gold-labeled p30 recombinant protein as the detection sensor, utilizing Staphylococcal Protein A (SPA) as the test line (T line), and p30 monoclonal antibody as the control line (C line). The sensitivity and specificity of the test strip were evaluated after optimizing the labeling concentration, pH, and protein dosage. The research findings revealed that the optimal colloidal gold labeling concentration was 0.05 %, the optimal pH was 8.4, and the optimal protein dosage was 10 μg/mL. Under these conditions, the CGITS demonstrated a detection limit of 1:512 dilution of ASFV standard positive serum, without exhibiting cross-reactivity with antibodies against other viral pathogens. Furthermore, the test strips remained stable for up to 20 days when stored at 50 °C and 4 °C. Comparatively, the CGITS outperformed commercial ELISA kits, displaying a sensitivity of 90.9 % and a specificity of 96.2 %. Subsequently, 108 clinical sera were tested to assess its performance. The data showed that the coincidence rate between the CGITS and ELISA was 93.5 %. In conclusion, the rapid colloidal gold test strip provides an efficient and reliable screening tool for on-site clinical detection of ASF in China. Its accuracy, stability, and simplicity make it a valuable asset in combating the spread of ASF and limiting its impact on the pig industry.
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spelling doaj.art-4e86f5c62db5408cb196a06056a646cf2024-02-17T06:40:04ZengElsevierHeliyon2405-84402024-02-01103e25214Development of high-concentration labeled colloidal gold immunochromatographic test strips for detecting african swine fever virus p30 protein antibodiesHuai-cheng Liu0Rong-chao Liu1Mei-rong Hu2Ao-bing Yang3Ren-hu Wu4Yan Chen5Jin Zhang6Ji-shan Bai7Sheng-bo Wu8Jian-peng Chen9Yun-feng Long10Yan Jiang11Bin Zhou12MOE Joint International Research Laboratory of Animal Health and Food Safety, College of Veterinary Medicine, Nanjing Agricultural University, Nanjing 210095, ChinaMOE Joint International Research Laboratory of Animal Health and Food Safety, College of Veterinary Medicine, Nanjing Agricultural University, Nanjing 210095, ChinaGuangDong Winsun Bio-Pharmaceutical Co., Ltd., Guangzhou, 511356, ChinaGuangDong Winsun Bio-Pharmaceutical Co., Ltd., Guangzhou, 511356, ChinaMOE Joint International Research Laboratory of Animal Health and Food Safety, College of Veterinary Medicine, Nanjing Agricultural University, Nanjing 210095, ChinaMOE Joint International Research Laboratory of Animal Health and Food Safety, College of Veterinary Medicine, Nanjing Agricultural University, Nanjing 210095, ChinaMOE Joint International Research Laboratory of Animal Health and Food Safety, College of Veterinary Medicine, Nanjing Agricultural University, Nanjing 210095, ChinaMOE Joint International Research Laboratory of Animal Health and Food Safety, College of Veterinary Medicine, Nanjing Agricultural University, Nanjing 210095, ChinaMOE Joint International Research Laboratory of Animal Health and Food Safety, College of Veterinary Medicine, Nanjing Agricultural University, Nanjing 210095, ChinaMOE Joint International Research Laboratory of Animal Health and Food Safety, College of Veterinary Medicine, Nanjing Agricultural University, Nanjing 210095, ChinaAnimal, Plant and Food Inspection Center, Nanjing Customs, Nanjing 210019, ChinaAnimal, Plant and Food Inspection Center, Nanjing Customs, Nanjing 210019, ChinaMOE Joint International Research Laboratory of Animal Health and Food Safety, College of Veterinary Medicine, Nanjing Agricultural University, Nanjing 210095, China; Corresponding author.African Swine Fever (ASF), caused by the African swine fever virus (ASFV), has inflicted significant economic losses on the pig industry in China. The key to mitigating its impact lies in accurate screening and strict biosecurity measures. In this regard, the development of colloidal gold immunochromatographic test strips (CGITS) has proven to be an effective method for detecting ASFV antibodies. These test strips are based on the ASFV p30 recombinant protein and corresponding monoclonal antibodies. The design of the test strip incorporates a high-concentration colloidal gold-labeled p30 recombinant protein as the detection sensor, utilizing Staphylococcal Protein A (SPA) as the test line (T line), and p30 monoclonal antibody as the control line (C line). The sensitivity and specificity of the test strip were evaluated after optimizing the labeling concentration, pH, and protein dosage. The research findings revealed that the optimal colloidal gold labeling concentration was 0.05 %, the optimal pH was 8.4, and the optimal protein dosage was 10 μg/mL. Under these conditions, the CGITS demonstrated a detection limit of 1:512 dilution of ASFV standard positive serum, without exhibiting cross-reactivity with antibodies against other viral pathogens. Furthermore, the test strips remained stable for up to 20 days when stored at 50 °C and 4 °C. Comparatively, the CGITS outperformed commercial ELISA kits, displaying a sensitivity of 90.9 % and a specificity of 96.2 %. Subsequently, 108 clinical sera were tested to assess its performance. The data showed that the coincidence rate between the CGITS and ELISA was 93.5 %. In conclusion, the rapid colloidal gold test strip provides an efficient and reliable screening tool for on-site clinical detection of ASF in China. Its accuracy, stability, and simplicity make it a valuable asset in combating the spread of ASF and limiting its impact on the pig industry.http://www.sciencedirect.com/science/article/pii/S2405844024012453African swine fever virus (ASFV)p30 proteinMonoclonal antibodyColloidal gold immunochromatographyHigh-concentration labelingTesting
spellingShingle Huai-cheng Liu
Rong-chao Liu
Mei-rong Hu
Ao-bing Yang
Ren-hu Wu
Yan Chen
Jin Zhang
Ji-shan Bai
Sheng-bo Wu
Jian-peng Chen
Yun-feng Long
Yan Jiang
Bin Zhou
Development of high-concentration labeled colloidal gold immunochromatographic test strips for detecting african swine fever virus p30 protein antibodies
Heliyon
African swine fever virus (ASFV)
p30 protein
Monoclonal antibody
Colloidal gold immunochromatography
High-concentration labeling
Testing
title Development of high-concentration labeled colloidal gold immunochromatographic test strips for detecting african swine fever virus p30 protein antibodies
title_full Development of high-concentration labeled colloidal gold immunochromatographic test strips for detecting african swine fever virus p30 protein antibodies
title_fullStr Development of high-concentration labeled colloidal gold immunochromatographic test strips for detecting african swine fever virus p30 protein antibodies
title_full_unstemmed Development of high-concentration labeled colloidal gold immunochromatographic test strips for detecting african swine fever virus p30 protein antibodies
title_short Development of high-concentration labeled colloidal gold immunochromatographic test strips for detecting african swine fever virus p30 protein antibodies
title_sort development of high concentration labeled colloidal gold immunochromatographic test strips for detecting african swine fever virus p30 protein antibodies
topic African swine fever virus (ASFV)
p30 protein
Monoclonal antibody
Colloidal gold immunochromatography
High-concentration labeling
Testing
url http://www.sciencedirect.com/science/article/pii/S2405844024012453
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