J Bacteriol Virol.  2016 Jun;46(2):63-70. 10.4167/jbv.2016.46.2.63.

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

• Lee SH
• Yang DK
• Kim HH
• Jo HY
• Choi SS
• Cho IS

• KMID: 2327901
• DOI: http://doi.org/10.4167/jbv.2016.46.2.63

Abstract

Getah virus (GETV) is a member of the genus Alphavirus in the family Togaviridae. GETV infection can occur in a wide range of vertebrate species, and the virus has been known for a pathogen of horses and pigs. To rapidly and accurately diagnose GETV infection of a racehorse, an indirect ELISA (I-ELISA) was developed in the present study for detection of antibodies to GETV in serum samples. To evaluate the developed I-ELISA, a total of 240 serum samples from Thoroughbred racehorses raised in Korea were screened in parallel by a serum neutralization (SN) test. The developed I-ELISA exhibited an efficacy comparable to that of the SN test in terms of a high diagnostic sensitivity (86.3%) and specificity (94.5%) at a cut-off absorbance value of 0.25. In addition, our results showed that the developed I-ELISA had a significant correlation with the SN test (r = 0.91; p < 0.05). Taken together, our findings suggest that the I-ELISA developed in this study is a valuable diagnostic tool for the screening of horses suspected to be infected with GETV.

Keyword

Getah virus; ELISA; Racehorse

MeSH Terms

Alphavirus*
Antibodies*
Enzyme-Linked Immunosorbent Assay*
Horses*
Humans
Korea
Mass Screening
Sensitivity and Specificity
Swine
Togaviridae
Vertebrates
Antibodies

Figure

• 

  Figure 1. The optimal time point for antigen harvest was determined by assessing GETV growth kinetics (A). The GETV-specific CPE was observed at ~24 hpi (B).

• 

  Figure 2. Checkerboard titration of horse serum against GETV antigen by I-ELISA. GETV-positive (SN titer of ≥ 1:64) and -negative (SN titer of < 1:2) reference sera were determined by the SN test. According to the checkerboard titration, the optimum working concentration of GETV antigen (A) and dilution of reference sera (B) were 2.5 μg/ml and 1:40, respectively. The senum samples were evaluated as positive if their OD value was greater than 0.25. This OD cut-off value was calculated as noted in the Materials and Methods.

• 

  Figure 3. Comparison of the SN test and I-ELlSA for detection of GETV antibodies in 240 horse serum samples. The correIation between the two tests is indicated by the linear regression line and r-value.

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