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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.


Receiver operating characteristic (ROC) for analysis of the cELISA. ROC curves for cELISA compared with four different IFA cut-off points. AUC, area under the curve.
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Figure 7: Receiver operating characteristic (ROC) for analysis of the cELISA. ROC curves for cELISA compared with four different IFA cut-off points. AUC, area under the curve.

Mentions: Because official standard methods for the antibody detection of SFTSV have not been determined, the cut-off criteria determination for IFA requires further optimization. Therefore, ROC curve was employed to determine the sensitivity and specificity and define the optimal cut-off value for IFA. The ROC curves of four different IFA cut-off values were compared, and the results showed that a 1/80 dilution of the sample strongly supports the result of cELISA. For this dilution, the area under the curve (AUC) was 0.91. While a 1/128 dilution rate as an IFA cut-off point maximizes the specificity and sensitivity compared with other dilution rates, this condition is too strict to detect positive samples above the strict threshold in cELISA. The other IFA cut-off conditions (1/16 and 1/64) were also excluded because they showed moderate AUC values (Fig. 7).


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
Receiver operating characteristic (ROC) for analysis of the cELISA. ROC curves for cELISA compared with four different IFA cut-off points. AUC, area under the curve.
© Copyright Policy - open-access
Related In: Results  -  Collection

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

Figure 7: Receiver operating characteristic (ROC) for analysis of the cELISA. ROC curves for cELISA compared with four different IFA cut-off points. AUC, area under the curve.
Mentions: Because official standard methods for the antibody detection of SFTSV have not been determined, the cut-off criteria determination for IFA requires further optimization. Therefore, ROC curve was employed to determine the sensitivity and specificity and define the optimal cut-off value for IFA. The ROC curves of four different IFA cut-off values were compared, and the results showed that a 1/80 dilution of the sample strongly supports the result of cELISA. For this dilution, the area under the curve (AUC) was 0.91. While a 1/128 dilution rate as an IFA cut-off point maximizes the specificity and sensitivity compared with other dilution rates, this condition is too strict to detect positive samples above the strict threshold in cELISA. The other IFA cut-off conditions (1/16 and 1/64) were also excluded because they showed moderate AUC values (Fig. 7).

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.