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Applied microbiology and biotechnology2024; 108(1); 355; doi: 10.1007/s00253-024-13168-5

Development and application of a colloidal-gold immunochromatographic strip for detecting Getah virus antibodies.

Abstract: Getah virus (GETV) is a re-emerging mosquito-borne alphavirus that is highly pathogenic, mainly to pigs and horses. There are no vaccines or treatments available for GETV in swine in China. Therefore, the development of a simple, rapid, specific, and sensitive serological assay for GETV antibodies is essential for the prevention and control of GETV. Current antibody monitoring methods are time-consuming, expensive, and dependent on specialized instrumentation, and these features are not conducive to rapid detection in clinical samples. To address these problem, we developed immunochromatographic test strips (ICTS) using eukaryotically expressed soluble recombinant p62-E1 protein of GETV as a labelled antigen, which has good detection sensitivity and no cross-reactivity with other common porcine virus-positive sera. The ICTS is highly compatible with IFA and ELISA and can be stored for 1 month at 37 °C and for at least 3 months at room temperature. Hence, p62-E1-based ICTS is a rapid, accurate, and convenient method for rapid on-site detection of GETV antibodies. KEY POINTS: • We established a rapid antibody detection method that can monitor GETV infection • We developed colloidal gold test strips with high sensitivity and specificity • The development of colloidal gold test strips will aid in the field serologic detection of GETV.
© 2024. The Author(s).
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Publication Date: 2024-06-01 PubMed ID: 38822832PubMed Central: PMC11144135DOI: 10.1007/s00253-024-13168-5Google Scholar: Lookup
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Summary

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Overview

  • This research focuses on developing a rapid, sensitive, and specific immunochromatographic strip test to detect antibodies against Getah virus (GETV) in animals, particularly pigs and horses.
  • The test aims to provide a practical solution for on-site screening and monitoring of GETV infections to aid in disease prevention and control.

Background

  • Getah virus (GETV) is a mosquito-borne alphavirus that is re-emerging and poses a significant pathogenic threat mainly to pigs and horses.
  • In regions like China, there are currently no vaccines or treatments available for GETV infections in swine populations.
  • Existing methods for detecting GETV antibodies, such as ELISA (Enzyme-Linked Immunosorbent Assay) and IFA (Immunofluorescence Assay), are accurate but:
    • Require specialized equipment and trained technicians.
    • Are time-consuming and costly.
    • Are difficult to implement in field or clinical settings for rapid diagnosis.

Objectives

  • To develop a simple, rapid, and cost-effective serological assay for the detection of GETV antibodies.
  • To create a test that can be used onsite, particularly in settings without advanced laboratory infrastructure.
  • To ensure the assay has high sensitivity and specificity, with no cross-reactivity to other porcine viruses.

Methodology

  • Utilized a recombinant version of the GETV p62-E1 protein expressed in eukaryotic cells, ensuring proper folding and antigenicity.
  • Conjugated the recombinant p62-E1 protein to colloidal gold particles to serve as the labeled antigen on immunochromatographic strips (ICTS).
  • Tested the ICTS strips against various clinical samples to evaluate:
    • Sensitivity for detecting GETV antibodies.
    • Specificity to ensure no cross-reactivity with antibodies from other common porcine viruses.
  • Evaluated compatibility by comparing results with established methods like IFA and ELISA.
  • Assessed the stability of the strips by storing them at 37°C for one month and at room temperature for at least three months.

Key Findings

  • The developed colloidal gold ICTS showed:
    • High sensitivity and specificity in detecting GETV antibodies.
    • No cross-reactivity with antibodies from other porcine viruses, indicating excellent specificity.
  • Results of the ICTS were consistent and highly compatible with traditional ELISA and IFA results.
  • The strips exhibited good stability, maintaining performance after storage at elevated temperatures and for several months at room temperature.
  • The assay provided results rapidly, making it ideal for point-of-care or field diagnostics.

Implications and Applications

  • The ICTS can be used for routine surveillance and monitoring of GETV infection in pig and horse populations.
  • Its rapid and simple operation allows veterinary practitioners or field workers to conduct testing without the need for sophisticated laboratory infrastructure.
  • The availability of this test will support early detection and aid in controlling the spread of GETV.
  • This technology could potentially be adapted or serve as a model for rapid detection assays for other emerging veterinary viral diseases.

Conclusion

  • The researchers successfully developed a colloidal gold immunochromatographic strip test based on the recombinant p62-E1 protein of GETV.
  • The test is fast, accurate, stable, and convenient, meeting the critical needs for on-site serological detection of GETV antibodies.
  • This assay represents an important advancement in GETV disease management with potential benefits for swine health and economic sustainability in affected regions.

Cite This Article

APA
Jiang Z, Qin Y, Zhang L, Xing G, Shi Z, Song W, Dobrikov GM, Chen J, Su S. (2024). Development and application of a colloidal-gold immunochromatographic strip for detecting Getah virus antibodies. Appl Microbiol Biotechnol, 108(1), 355. https://doi.org/10.1007/s00253-024-13168-5

Publication

Applied microbiology and biotechnology
ISSN: 1432-0614
NlmUniqueID: 8406612
Country: Germany
Language: English
Volume: 108
Issue: 1
Pages: 355
PII: 355

Researcher Affiliations

Jiang, Zhiwen
  • Jiangsu Engineering Laboratory of Animal Immunology, Institute of Immunology and College of Veterinary Medicine, Nanjing Agricultural University, Nanjing, 210095, China.
  • Sanya Institute of Nanjing Agricultural University, Sanya, China.
Qin, Ying
  • Jiangsu Engineering Laboratory of Animal Immunology, Institute of Immunology and College of Veterinary Medicine, Nanjing Agricultural University, Nanjing, 210095, China.
  • Sanya Institute of Nanjing Agricultural University, Sanya, China.
Zhang, Letian
  • Jiangsu Engineering Laboratory of Animal Immunology, Institute of Immunology and College of Veterinary Medicine, Nanjing Agricultural University, Nanjing, 210095, China.
Xing, Gang
  • MOA Key Laboratory of Animal Virology, Zhejiang University, Hangzhou, 310058, China.
Shi, Zhiyu
  • Jiangsu Engineering Laboratory of Animal Immunology, Institute of Immunology and College of Veterinary Medicine, Nanjing Agricultural University, Nanjing, 210095, China.
Song, Wanjie
  • Jiangsu Engineering Laboratory of Animal Immunology, Institute of Immunology and College of Veterinary Medicine, Nanjing Agricultural University, Nanjing, 210095, China.
Dobrikov, Georgi M
  • Institute of Organic Chemistry With Centre of Phytochemistry, Bulgarian Academy of Sciences, Acad. G. Bonchev Street, Bl. 9, 1113, Sofia, Bulgaria.
Chen, Jie
  • Jiangsu Engineering Laboratory of Animal Immunology, Institute of Immunology and College of Veterinary Medicine, Nanjing Agricultural University, Nanjing, 210095, China.
Su, Shuo
  • Jiangsu Engineering Laboratory of Animal Immunology, Institute of Immunology and College of Veterinary Medicine, Nanjing Agricultural University, Nanjing, 210095, China. shuosu@njau.edu.cn.
  • Sanya Institute of Nanjing Agricultural University, Sanya, China. shuosu@njau.edu.cn.

MeSH Terms

  • Animals
  • Gold Colloid / chemistry
  • Antibodies, Viral / blood
  • Antibodies, Viral / immunology
  • Alphavirus / immunology
  • Swine
  • Sensitivity and Specificity
  • Chromatography, Affinity / methods
  • Alphavirus Infections / diagnosis
  • Alphavirus Infections / immunology
  • Swine Diseases / diagnosis
  • Swine Diseases / virology
  • Reagent Strips
  • China
  • Enzyme-Linked Immunosorbent Assay / methods

Conflict of Interest Statement

The authors declare no competing interests.

References

This article includes 26 references
  1. Bai Y, Jia R, Wei Q, Wang L, Sun Y, Li Y, Luo J, Zhang G. Development and application of a high-sensitivity immunochromatographic test strip for detecting classical swine fever virus antibodies. Transbound Emerg Dis 69(4):e788–e798.
    doi: 10.1111/tbed.14367pubmed: 34724351google scholar: lookup
  2. Cao L, Kong X, Li X, Suo X, Duan Y, Yuan C, Zhang Y, Zheng H, Wang Q. A customized novel blocking ELISA for detection of bat-origin swine acute diarrhea syndrome coronavirus infection. Microbiol Spectr 11(4):e0393022.
    doi: 10.1128/spectrum.03930-22pmc: PMC10434073pubmed: 37272819google scholar: lookup
  3. Cavalera S, Colitti B, Rosati S, Ferrara G, Bertolotti L, Nogarol C, Guiotto C, Cagnazzo C, Denina M, Fagioli F, Di Nardo F, Chiarello M, Baggiani C, Anfossi L. A multi-target lateral flow immunoassay enabling the specific and sensitive detection of total antibodies to SARS COV-2. Talanta 223(Pt 1):121737.
    doi: 10.1016/j.talanta.2020.121737pmc: PMC7534878pubmed: 33303174google scholar: lookup
  4. Kim AS, Diamond MS. A molecular understanding of alphavirus entry and antibody protection. Nat Rev Microbiol 21(6):396–407.
    doi: 10.1038/s41579-022-00825-7pmc: PMC9734810pubmed: 36474012google scholar: lookup
  5. Kim AS, Kafai NM, Winkler ES, Gilliland TC Jr, Cottle EL, Earnest JT, Jethva PN, Kaplonek P, Shah AP, Fong RH, Davidson E, Malonis RJ, Quiroz JA, Williamson LE, Vang L, Mack M, Crowe JE Jr, Doranz BJ, Lai JR, Alter G, Gross ML, Klimstra WB, Fremont DH, Diamond MS. Pan-protective anti-alphavirus human antibodies target a conserved E1 protein epitope. Cell 184(17):4414-4429.e19.
    doi: 10.1016/j.cell.2021.07.006pmc: PMC8382027pubmed: 34416146google scholar: lookup
  6. Li YY, Liu H, Fu SH, Li XL, Guo XF, Li MH, Feng Y, Chen WX, Wang LH, Lei WW, Gao XY, Lv Z, He Y, Wang HY, Zhou HN, Wang GQ, Liang GD. From discovery to spread: the evolution and phylogeny of Getah virus. Infect Genet Evol 55:48–55.
    doi: 10.1016/j.meegid.2017.08.016pubmed: 28827175google scholar: lookup
  7. Li G, Li Q, Wang X, Liu X, Zhang Y, Li R, Guo J, Zhang G. Lateral flow immunoassays for antigens, antibodies and haptens detection. Int J Biol Macromol 242(Pt 4):125186.
    doi: 10.1016/j.ijbiomac.2023.125186pmc: PMC10232721pubmed: 37268073google scholar: lookup
  8. Li B, Wang H, Liang G. Getah Virus (Alphavirus): an emerging, spreading zoonotic virus. Pathogens 11(8).
    pmc: PMC9416625pubmed: 36015065doi: 10.3390/pathogens11080945google scholar: lookup
  9. Liu H, Zhang X, Li LX, Shi N, Sun XT, Liu Q, Jin NY, Si XK. First isolation and characterization of Getah virus from cattle in northeastern China. BMC Vet Res 15(1):320.
    doi: 10.1186/s12917-019-2061-zpmc: PMC6729113pubmed: 31488162google scholar: lookup
  10. Lu G, Ou J, Ji J, Ren Z, Hu X, Wang C, Li S. Emergence of Getah virus infection in horse with fever in China, 2018. Front Microbiol 10:1416.
    doi: 10.3389/fmicb.2019.01416pmc: PMC6596439pubmed: 31281304google scholar: lookup
  11. Malonis RJ, Earnest JT, Kim AS, Angeliadis M, Holtsberg FW, Aman MJ, Jangra RK, Chandran K, Daily JP, Diamond MS, Kielian M, Lai JR. Near-germline human monoclonal antibodies neutralize and protect against multiple arthritogenic alphaviruses. Proc Natl Acad Sci U S A 118(37).
    pmc: PMC8449321pubmed: 34507983doi: 10.1073/pnas.2100104118google scholar: lookup
  12. Martins KA, Gregory MK, Valdez SM, Sprague TR, Encinales L, Pacheco N, Cure C, Porras-Ramirez A, Rico-Mendoza A, Chang A, Pitt ML, Nasar F. Neutralizing antibodies from convalescent chikungunya virus patients can cross-neutralize Mayaro and Una viruses. Am J Trop Med Hyg 100(6):1541–1544.
    doi: 10.4269/ajtmh.18-0756pmc: PMC6553899pubmed: 31017081google scholar: lookup
  13. Nguyen W, Nakayama E, Yan K, Tang B, Le TT, Liu L, Cooper TH, Hayball JD, Faddy HM, Warrilow D, Allcock RJN, Hobson-Peters J, Hall RA, Rawle DJ, Lutzky VP, Young P, Oliveira NM, Hartel G, Howley PM, Prow NA, Suhrbier A. Arthritogenic alphavirus vaccines: serogrouping versus cross-protection in mouse models. Vaccines (Basel) 8(2).
    pmc: PMC7349283pubmed: 32380760doi: 10.3390/vaccines8020209google scholar: lookup
  14. Qiu X, Cao X, Shi N, Zhang H, Zhu X, Gao Y, Mai Z, Jin N, Lu H. Development and application of an indirect ELISA for detecting equine IgG antibodies against Getah virus with recombinant E2 domain protein. Front Microbiol 13:1029444.
    doi: 10.3389/fmicb.2022.1029444pmc: PMC9685671pubmed: 36439788google scholar: lookup
  15. Shi N, Li LX, Lu RG, Yan XJ, Liu H. Highly pathogenic swine Getah virus in blue foxes, Eastern China, 2017. Emerg Infect Dis 25(6):1252–1254.
    doi: 10.3201/eid2506.181983pmc: PMC6537705pubmed: 31107236google scholar: lookup
  16. Sun Q, Xie Y, Guan Z, Zhang Y, Li Y, Yang Y, Zhang J, Li Z, Qiu Y, Li B, Liu K, Shao D, Wang J, Ma Z, Wei J, Li P. Seroprevalence of Getah virus in pigs in Eastern China determined with a recombinant E2 protein-based indirect ELISA. Viruses 14(10).
    pmc: PMC9607375pubmed: 36298726doi: 10.3390/v14102173google scholar: lookup
  17. Uchime O, Fields W, Kielian M. The role of E3 in pH protection during alphavirus assembly and exit. J Virol 87(18):10255–10262.
    doi: 10.1128/jvi.01507-13pmc: PMC3754015pubmed: 23864626google scholar: lookup
  18. Voss JE, Vaney MC, Duquerroy S, Vonrhein C, Girard-Blanc C, Crublet E, Thompson A, Bricogne G, Rey FA. Glycoprotein organization of Chikungunya virus particles revealed by X-ray crystallography. Nature 468(7324):709–712.
    doi: 10.1038/nature09555pubmed: 21124458google scholar: lookup
  19. Wang A, Zhou F, Liu C, Gao D, Qi R, Yin Y, Liu S, Gao Y, Fu L, Xia Y, Xu Y, Wang C, Liu Z. Structure of infective Getah virus at 2.8 Å resolution determined by cryo-electron microscopy. Cell Discov 8(1):12.
    doi: 10.1038/s41421-022-00374-6pmc: PMC8832435pubmed: 35149682google scholar: lookup
  20. Wang N, Zhai X, Li X, Wang Y, He WT, Jiang Z, Veit M, Su S. Attenuation of Getah virus by a single amino acid substitution at residue 253 of the E2 protein that might be part of a new heparan sulfate binding site on alphaviruses. J Virol 96(6):e0175121.
    doi: 10.1128/jvi.01751-21pmc: PMC8941864pubmed: 34986000google scholar: lookup
  21. Williamson LE, Reeder KM, Bailey K, Tran MH, Roy V, Fouch ME, Kose N, Trivette A, Nargi RS, Winkler ES, Kim AS, Gainza C, Rodriguez J, Armstrong E, Sutton RE, Reidy J, Carnahan RH, McDonald WH, Schoeder CT, Klimstra WB, Davidson E, Doranz BJ, Alter G, Meiler J, Schey KL, Julander JG, Diamond MS, Crowe JE Jr. Therapeutic alphavirus cross-reactive E1 human antibodies inhibit viral egress. Cell 184(17):4430-4446.e22.
    doi: 10.1016/j.cell.2021.07.033pmc: PMC8418820pubmed: 34416147google scholar: lookup
  22. Yang T, Li R, Hu Y, Yang L, Zhao D, Du L, Li J, Ge M, Yu X. An outbreak of Getah virus infection among pigs in China, 2017. Transbound Emerg Dis 65(3):632–637.
    doi: 10.1111/tbed.12867pubmed: 29575687google scholar: lookup
  23. Zhang R, Kim AS, Fox JM, Nair S, Basore K, Klimstra WB, Rimkunas R, Fong RH, Lin H, Poddar S, Crowe JE Jr, Doranz BJ, Fremont DH, Diamond MS. Mxra8 is a receptor for multiple arthritogenic alphaviruses. Nature 557(7706):570–574.
    doi: 10.1038/s41586-018-0121-3pmc: PMC5970976pubmed: 29769725google scholar: lookup
  24. Zhang Y, Li Y, Guan Z, Yang Y, Zhang J, Sun Q, Li B, Qiu Y, Liu K, Shao D, Ma Z, Wei J, Li P. Rapid differential detection of Japanese encephalitis virus and Getah virus in pigs or mosquitos by a Duplex TaqMan real-time RT-PCR assay. Front Vet Sci 9:839443.
    doi: 10.3389/fvets.2022.839443pmc: PMC9023051pubmed: 35464361google scholar: lookup
  25. Zhao M, Yue C, Yang Z, Li Y, Zhang D, Zhang J, Yang S, Shen Q, Su X, Qi D, Ma R, Xiao Y, Hou R, Yan X, Li L, Zhou Y, Liu J, Wang X, Wu W, Zhang W, Shan T, Liu S. Viral metagenomics unveiled extensive communications of viruses within giant pandas and their associated organisms in the same ecosystem. Sci Total Environ 820:153317.
    doi: 10.1016/j.scitotenv.2022.153317pubmed: 35066043google scholar: lookup
  26. Zhao J, Dellicour S, Yan Z, Veit M, Gill MS, He WT, Zhai X, Ji X, Suchard MA, Lemey P, Su S. Early genomic surveillance and phylogeographic analysis of Getah virus, a reemerging arbovirus, in livestock in China. J Virol 97(1):e0109122.
    doi: 10.1128/jvi.01091-22pmc: PMC9888209pubmed: 36475767google scholar: lookup

Citations

This article has been cited 2 times.
  1. Ling X-h, Ruan W-y, Ren H-j, Zhang B, Cong J-h, Yuan W-j, Liu G-s, Huang H-h, Chen Z-j, Han S-c, He W-r, Zhang G-p, Zhang Y-h, Wan B. A novel 3A-based indirect enzyme-linked immunosorbent assay for serological testing of Senecavirus A.. Microbiol Spectr 2025 Dec 2;13(12):e0077925.
    doi: 10.1128/spectrum.00779-25pubmed: 41165357google scholar: lookup
  2. Jiang Z, Merits A, Qin Y, Xing G, Zhang L, Chen J, Wang N, Varjak M, Zhai X, Li D, Song W, Su S. Attenuated Getah virus confers protection against multiple arthritogenic alphaviruses.. PLoS Pathog 2024 Nov;20(11):e1012700.
    doi: 10.1371/journal.ppat.1012700pubmed: 39556619google scholar: lookup
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