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Development and evaluation of a competitive enzyme-linked immunosorbent assay using a monoclonal antibody for diagnosis of severe fever with thrombocytopenia syndrome virus in bovine sera

View Article: PubMed Central - PubMed

ABSTRACT

Severe fever with thrombocytopenia syndrome (SFTS) caused by the SFTS virus (SFTSV), a phlebovirus in the family Bunyaviridae, is an emerging tick-borne infectious disease that impacts humans. This disease manifests as a decreased blood cell count and multi-organ failure, with a case-fatality rate of more than 12% in China. Because vaccines or antiviral drugs for the treatment of this disease are not available, monitoring the SFTS circulation in animals and controlling the tick-mammal cycle are important for preventing SFTS. Monoclonal antibodies against the recombinant nucleoprotein of SFTSV were generated to develop a competitive enzyme-linked immunosorbent assay (cELISA) for the detection of antibodies against SFTSV infection in cattle. The specificity and sensitivity of cELISA was assessed by comparing the results of this assay to those of an immunofluorescence assay (IFA). The results of the cELISA using 416 field bovine serum samples and laboratory-immunized positive sera showed 98.1% consistency with those of the IFA. The cELISA used in this study did not show cross-reactivity with antisera against other viral cattle diseases. The cELISA presented in this study can be applied to detect antibodies against SFTSV in cattle.

No MeSH data available.


Establishment of cut-off PI value in cELISA. Frequency distributions of the PI values for 407 bovine serum samples determined as SFTSV-negative in IFA (1 : 80 dilution). The cut-off of 49.5% (mean ± 3SD) is indicated with the arrow.
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Figure 5: Establishment of cut-off PI value in cELISA. Frequency distributions of the PI values for 407 bovine serum samples determined as SFTSV-negative in IFA (1 : 80 dilution). The cut-off of 49.5% (mean ± 3SD) is indicated with the arrow.

Mentions: A total of 416 bovine serum samples and two positive control sera were tested with the IFA to distinguish negative serum samples from SFTSV. The results showed that 97.4% (407/418) of the bovine serum samples were negative, while 2.6% (11/418) of the bovine serum samples were positive according to the IFA. cELISA was performed using the SFTS-negative group samples, and the mean inhibition percent was 11.3% with a standard deviation of 12.7%. Therefore, we determined the cut-off value to be 49.5% (mean ± 3SD) (Fig. 5). The cELISA and IFA were compared using this cut-off value, and 98.1% (410/418) consistency was observed between results (Table 1). The accuracy of cELISA was demonstrated with a plot of the distribution of PI values obtained from the IFA-negative or positive cattle group (Fig. 6).


Development and evaluation of a competitive enzyme-linked immunosorbent assay using a monoclonal antibody for diagnosis of severe fever with thrombocytopenia syndrome virus in bovine sera
Establishment of cut-off PI value in cELISA. Frequency distributions of the PI values for 407 bovine serum samples determined as SFTSV-negative in IFA (1 : 80 dilution). The cut-off of 49.5% (mean ± 3SD) is indicated with the arrow.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC5037297&req=5

Figure 5: Establishment of cut-off PI value in cELISA. Frequency distributions of the PI values for 407 bovine serum samples determined as SFTSV-negative in IFA (1 : 80 dilution). The cut-off of 49.5% (mean ± 3SD) is indicated with the arrow.
Mentions: A total of 416 bovine serum samples and two positive control sera were tested with the IFA to distinguish negative serum samples from SFTSV. The results showed that 97.4% (407/418) of the bovine serum samples were negative, while 2.6% (11/418) of the bovine serum samples were positive according to the IFA. cELISA was performed using the SFTS-negative group samples, and the mean inhibition percent was 11.3% with a standard deviation of 12.7%. Therefore, we determined the cut-off value to be 49.5% (mean ± 3SD) (Fig. 5). The cELISA and IFA were compared using this cut-off value, and 98.1% (410/418) consistency was observed between results (Table 1). The accuracy of cELISA was demonstrated with a plot of the distribution of PI values obtained from the IFA-negative or positive cattle group (Fig. 6).

View Article: PubMed Central - PubMed

ABSTRACT

Severe fever with thrombocytopenia syndrome (SFTS) caused by the SFTS virus (SFTSV), a phlebovirus in the family Bunyaviridae, is an emerging tick-borne infectious disease that impacts humans. This disease manifests as a decreased blood cell count and multi-organ failure, with a case-fatality rate of more than 12% in China. Because vaccines or antiviral drugs for the treatment of this disease are not available, monitoring the SFTS circulation in animals and controlling the tick-mammal cycle are important for preventing SFTS. Monoclonal antibodies against the recombinant nucleoprotein of SFTSV were generated to develop a competitive enzyme-linked immunosorbent assay (cELISA) for the detection of antibodies against SFTSV infection in cattle. The specificity and sensitivity of cELISA was assessed by comparing the results of this assay to those of an immunofluorescence assay (IFA). The results of the cELISA using 416 field bovine serum samples and laboratory-immunized positive sera showed 98.1% consistency with those of the IFA. The cELISA used in this study did not show cross-reactivity with antisera against other viral cattle diseases. The cELISA presented in this study can be applied to detect antibodies against SFTSV in cattle.

No MeSH data available.