Development and Evaluation of Indirect ELISA for Detection of Antibodies to Getah Virus in Horse Serum

Lee, Seung Heon; Yang, Dong-Kun; Kim, Ha-Hyun; Jo, Hyun-Ye; Choi, Sung-Suk; Cho, In Soo

doi:10.4167/jbv.2016.46.2.63

Cited by 6 publications

(2 citation statements)

References 27 publications

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“…Even so, there is still no efficient commercial diagnostic kit for GETV antibodies, leading to serious inconvenience to both carry out epidemiological investigation of GETV disease and further strengthen the prevention and control of GETV disease. Taking ELISA’s advantages of sensitivity and suitability for the detection of large quantities of samples, some studies have reported that ELISA methods for detecting GETV antibody derived from pigs and horses could be established successfully using purified GETV virion as antigens ( Lee et al, 2016 ). However, this method of preparing antigens utilizing virion may face the bio-safety risk of virus dispersal.…”

Section: Discussionmentioning

confidence: 99%

Development and application of an indirect ELISA for detecting equine IgG antibodies against Getah virus with recombinant E2 domain protein

Qiu

Cao²,

Shi³

et al. 2022

Front. Microbiol.

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Getah virus (GETV) disease is a mosquito-borne infectious disease that causes fever, aseptic meningitis, and abortion in a variety of animals. Currently, the epidemic trend of GETV disease increases seriously worldwide, especially in China, posing a potential threat to animal safety and public health. However, there are few reports about the epidemiological investigation of GETV disease in China as well as a lack of commercial diagnostic kit for GETV antibody. Therefore, the establishment of a rapid, sensitive and suitable GETV antibody detection method for large-scale samples is an urgent request to fully understand the prevalence of GETV disease. Here, a recombinant plasmid pET22b-GETV-E2d that contained the domain of GETV-E2 (E2d) fused to His-tag was constructed to express recombinant protein E2d (rE2d) in Escherichia coli. The rE2d was mainly expressed in inclusion bodies. And it was purified successfully by nickel affinity column so that it could be used to develop an indirect ELISA (rE2d-ELISA). After optimizing reaction conditions of rE2d-ELISA, the cut-off value was determined as 0.396 with 100 equine sera tested by virus neutralization test (VNT). Furthermore, rE2d-ELISA method showed the positive rate of IgG antibodies against GETV was 54.3% based on testing 646 clinical serum samples obtained in Xinjiang whereas the overall coincidence rate between rE2d-ELISA and VNT was 94.0%, with 98.2% sensitivity and 92.6% specificity. The findings suggest that the developed IgG ELISA employing recombinant E2d promises was an efficient and low-cost type of antibody detection method for horse, which will benefit for prevention of GETV outbreaks in horses.

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Section: Discussionmentioning

confidence: 99%

Development and application of an indirect ELISA for detecting equine IgG antibodies against Getah virus with recombinant E2 domain protein

Qiu

Cao²,

Shi³

et al. 2022

Front. Microbiol.

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“…Previously, we purified GETV using the sucrose gradient method and reported an I-ELISA with sensitivity (86.3%) and high specificity (94.5%) compared with that of the VN test (16). In this study, we developed a blocking ELISA (B-ELISA) with high sensitivity and specificity that can be applied to several animal species because GETV transmitted by mosquitoes can cause disease in several animals.…”

Section: Introductionmentioning

confidence: 99%

Development of a Blocking ELISA Method for Detection of Getah Virus Antibodies in Horse Sera

Yang,

Park,

Kim

et al. 2023

J Bacteriol Virol.

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Recent climate change and global warming are likely to increase mosquito-borne diseases. Getah virus (GETV) is a mosquito-borne virus that infects horses and other mammals, including humans. Currently, GETV infection in horse sera is confirmed by the virus neutralization (VN) test. Therefore, there is a need for a new enzyme-linked immunosorbent assay (ELISA) to detect GETV-antibody in a large number of horse serum samples. We aimed to develop a blocking ELISA (B-ELISA) method for the specific detection of GETV antibodies in horse serum samples. Antibodies against GETV in the sera of 175 horses were measured using the VN test. The purified QIAG9301-100P virus was used as an antigen for the B-ELISA. A monoclonal antibody (1E1) was conjugated with horseradish peroxidase and used as a detection antibody. For the establishment of the B-ELISA, antigen concentration, serum dilution factor, and conjugate dilution concentration were determined as 5 µg/mL, 20 times, and 3.6 µg/mL, respectively. We evaluated the sensitivity, specificity, and accuracy of the B-ELISA using sera from 175 horses. The B-ELISA had a diagnostic sensitivity of 91.4%, a specificity of 94.0%, and an accuracy of 93.1% compared with that of the VN test. The B-ELISA was significantly correlated with the VN test (r = 0.83). The new B-ELISA could replace the VN test and be useful for the sero-surveillance of GETV in horse sera. It is also expected to be able to detect GETV antibodies in the serum of various animals including pigs.

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Establishment and application of an indirect ELISA for Getah virus E2 antibody detection

You,

Wang,

et al. 2024

Journal of Virological Methods

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Development and Evaluation of Indirect ELISA for Detection of Antibodies to Getah Virus in Horse Serum

Cited by 6 publications

References 27 publications

Development and application of an indirect ELISA for detecting equine IgG antibodies against Getah virus with recombinant E2 domain protein

Development and application of an indirect ELISA for detecting equine IgG antibodies against Getah virus with recombinant E2 domain protein

Development of a Blocking ELISA Method for Detection of Getah Virus Antibodies in Horse Sera

Establishment and application of an indirect ELISA for Getah virus E2 antibody detection

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