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Applied microbiology and biotechnology2024; 108(1); 1-13; doi: 10.1007/s00253-023-12980-9

Development and evaluation of a test strip for the rapid detection of antibody against equine infectious anemia virus.

Abstract: Equine infectious anemia (EIA) is a contagious disease of horses caused by the equine infectious anemia virus (EIAV). The clinical signs at the acute phase include intermittent high fever, thrombocytopenia, hemorrhage, edema, and anemia. The clinical signs at chronic and relapsing subclinical levels include emaciation and progressive weakness. Surviving horses become lifelong carriers because of the integration of the viral genome into that of the host, and these horses can produce and transmit the virus to other animals. This increases the difficulty of imposing practical control measures to prevent epidemics of this disease. Serological tests measuring the antibodies in equine sera are considered to be a reliable tool for the long-term monitoring of EIA. However, the standard serological tests for EIV either have low sensitivity (e.g., agar gel immunodiffusion test, AGID) or are time consuming to perform (e.g., ELISA and western blotting). The development of a rapid and simple method for detecting the disease is therefore critical to control the spread of EIA. In this study, we designed and developed a colloidal gold immunochromatographic (GICG) test strip to detect antibodies against EIAV based on the double-antigen sandwich. Both the p26 and gp45 proteins were used as the capture antigens, which may help to improve the positive detection rate of the strip. We found that the sensitivity of the test strip was 8 to 16 times higher than those of two commercially available ELISA tests and 128 to 256 times higher than AGID, but 8 to 16 times lower than that of western blotting. The strip has good specificity and stability. When serum samples from experimental horses immunized with the attenuated EIAV vaccine (n = 31) were tested, the results of the test strip showed 100% coincidence with those from NECVB-cELISA and 70.97% with AGID. When testing clinical serum samples (n = 1014), the test strip surprisingly provided greater sensitivity and a higher number of "true positive" results than other techniques. Therefore, we believe that the GICG test strip has demonstrated great potential in the field trials as a simple and effective tool for the detection of antibodies against EIAV. KEY POINTS: • A colloidal gold immunochromatographic (GICG) fast test strip was developed with good specificity, sensitivity, stability, and repeatability • The test strip can be used in point-of-care testing for the primary screening of EIAV antibodies • Both the p26 and gp45 proteins were used as the capture antigens, giving a high positive detection rate in the testing of experimentally infected animal and field samples.
© 2024. The Author(s).
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Publication Date: 2024-01-08 PubMed ID: 38189948PubMed Central: 2168377DOI: 10.1007/s00253-023-12980-9Google Scholar: Lookup
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Summary

This research summary has been generated with artificial intelligence and may contain errors and omissions. Refer to the original study to confirm details provided. Submit correction.

The research presents the development and evaluation of a test strip that can quickly detect antibodies against the equine infectious anemia virus. This fast-acting test strip has high sensitivity and specificity, making it a powerful tool for preventing the spread of the disease by identifying carriers early on.

Development of the Test Strip

  • The researchers aimed to create a test strip for the rapid detection of equine infectious ania virus (EIAV) in response to shortcomings with current serological tests. The older methods were either time-consuming or lacked sensitivity.
  • For this purpose, they designed a colloidal gold immunochromatographic (GICG) test strip which uses a double-antigen sandwich for detection.
  • The antigens used for capturing in this test were p26 and gp45 proteins, which were expected to enhance the positive detection rate of the strip.

Evaluation of the Test Strip

  • The sensitivity of the developed test strip was assessed and compared with other techniques. It proved 8 to 16 times more sensitive than commercial ELISA tests, and 128 to 256 times more than the Agar gel immunodiffusion test (AGID). However, it was 8 to 16 times less sensitive than western blotting.
  • Test strip’s specificity and stability were adjudged to be good. The experimentation with serum samples from horses vaccinated with the attenuated EIAV showed 100% correlation with NECVB-cELISA results and 70.97% with AGID.
  • When used to test clinical serum samples (n = 1014), the test strip was surprisingly superior in sensitivity and provided more true positive results than other techniques used.

Conclusions

  • The developed GICG test strip showed strong potential in field trials. It was recognized as a straightforward and effective tool for the detection of EIAV antibodies.
  • The researchers conclude that this test strip could be a worthwhile asset for primary screening of EIAV antibodies in point-of-care testing.
  • Using both p26 and gp45 proteins as capture antigens proved успешно, delivering a high positive detection rate in the experimental infection tests and field samples.

Cite This Article

APA
Zhang Z, Guo K, Chu X, Liu M, Du C, Hu Z, Wang X. (2024). Development and evaluation of a test strip for the rapid detection of antibody against equine infectious anemia virus. Appl Microbiol Biotechnol, 108(1), 1-13. https://doi.org/10.1007/s00253-023-12980-9

Publication

Applied microbiology and biotechnology
ISSN: 1432-0614
NlmUniqueID: 8406612
Country: Germany
Language: English
Volume: 108
Issue: 1
Pages: 1-13

Researcher Affiliations

Zhang, Zenan
  • State Key Laboratory for Animal Disease Control and Prevention, Harbin Veterinary Research Institute, the Chinese Academy of Agricultural Sciences, Harbin, China.
Guo, Kui
  • State Key Laboratory for Animal Disease Control and Prevention, Harbin Veterinary Research Institute, the Chinese Academy of Agricultural Sciences, Harbin, China.
Chu, Xiaoyu
  • State Key Laboratory for Animal Disease Control and Prevention, Harbin Veterinary Research Institute, the Chinese Academy of Agricultural Sciences, Harbin, China.
  • Institute of Western Agriculture, the Chinese Academy of Agricultural Sciences, Changji, China.
  • WOAH Reference Laboratory for Equine Infectious Anemia, Harbin Veterinary Research Institute, the Chinese Academy of Agricultural Sciences, Harbin, China.
Liu, Mingru
  • Shenzhen Lvshiyuan Biotechnology Co., Shenzhen, China.
Du, Cheng
  • State Key Laboratory for Animal Disease Control and Prevention, Harbin Veterinary Research Institute, the Chinese Academy of Agricultural Sciences, Harbin, China. ducheng@caas.cn.
  • WOAH Reference Laboratory for Equine Infectious Anemia, Harbin Veterinary Research Institute, the Chinese Academy of Agricultural Sciences, Harbin, China. ducheng@caas.cn.
Hu, Zhe
  • State Key Laboratory for Animal Disease Control and Prevention, Harbin Veterinary Research Institute, the Chinese Academy of Agricultural Sciences, Harbin, China. huzher@126.com.
  • WOAH Reference Laboratory for Equine Infectious Anemia, Harbin Veterinary Research Institute, the Chinese Academy of Agricultural Sciences, Harbin, China. huzher@126.com.
Wang, Xiaojun
  • State Key Laboratory for Animal Disease Control and Prevention, Harbin Veterinary Research Institute, the Chinese Academy of Agricultural Sciences, Harbin, China. wangxiaojun@caas.cn.
  • Institute of Western Agriculture, the Chinese Academy of Agricultural Sciences, Changji, China. wangxiaojun@caas.cn.
  • WOAH Reference Laboratory for Equine Infectious Anemia, Harbin Veterinary Research Institute, the Chinese Academy of Agricultural Sciences, Harbin, China. wangxiaojun@caas.cn.

MeSH Terms

  • Animals
  • Horses
  • Infectious Anemia Virus, Equine
  • Antibodies, Viral
  • Enzyme-Linked Immunosorbent Assay
  • Blotting, Western
  • Gold Colloid

Grant Funding

  • 2021YFD1800500 / The National Key Research and Development Program of China
  • 2022YFD1800200 / The National Key Research and Development Program of China
  • TD2022C006 / The Natural Science Foundation of Heilongjiang Province of China

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Citations

This article has been cited 4 times.
  1. Liang H, Zhou B, Hu Z, Chu X, Wang X, Du C, Wang X. Development of a Broad-Spectrum Antigen-Capture ELISA Using Combined Anti-p26 Polyclonal and Monoclonal Antibodies for Detection of Equine Infectious Anemia Virus. Microorganisms 2025 Jun 27;13(7).
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  2. Firdausy LW, Fikri F, Wicaksono AP, Çalışkan H, Purnama MTE. Global prevalence and risk factors of equine infectious anemia: A systematic review and meta-analysis. Vet World 2025 Jun;18(6):1440-1451.
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