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Spot the difference-development of a syndrome based protein microarray for specific serological detection of multiple flavivirus infections in travelers.

Cleton NB, Godeke GJ, Reimerink J, Beersma MF, Doorn HR, Franco L, Goeijenbier M, Jimenez-Clavero MA, Johnson BW, Niedrig M, Papa A, Sambri V, Tami A, Velasco-Salas ZI, Koopmans MP, Reusken CB - PLoS Negl Trop Dis (2015)

Bottom Line: Sera from clinical flavivirus patients were used for primary development of the protein microarray.Results show a high IgG and IgM sensitivity and specificity for individual NS1 antigens, and limited cross reactivity, even within serocomplexes.Based on these data, multiplex NS1-based protein microarray is a promising tool for surveillance and diagnosis of flaviviruses.

View Article: PubMed Central - PubMed

Affiliation: Erasmus Medical Centre, Viroscience Department, Rotterdam, The Netherlands; National Institute for Public Health and Environment, Center for Infectious Diseases Research and Screening, Bilthoven, The Netherlands.

ABSTRACT

Background: The family Flaviviridae, genus Flavivirus, holds many of the world's most prevalent arboviral diseases that are also considered the most important travel related arboviral infections. In most cases, flavivirus diagnosis in travelers is primarily based on serology as viremia is often low and typically has already been reduced to undetectable levels when symptoms set in and patients seek medical attention. Serological differentiation between flaviviruses and the false-positive results caused by vaccination and cross-reactivity among the different species, are problematic for surveillance and diagnostics of flaviviruses. Their partially overlapping geographic distribution and symptoms, combined with increase in travel, and preexisting antibodies due to flavivirus vaccinations, expand the need for rapid and reliable multiplex diagnostic tests to supplement currently used methods.

Goal: We describe the development of a multiplex serological protein microarray using recombinant NS1 proteins for detection of medically important viruses within the genus Flavivirus. Sera from clinical flavivirus patients were used for primary development of the protein microarray.

Results: Results show a high IgG and IgM sensitivity and specificity for individual NS1 antigens, and limited cross reactivity, even within serocomplexes. In addition, the serology based on this array allows for discrimination between infection and vaccination response for JEV vaccine, and no cross-reactivity with TBEV and YFV vaccine induced antibodies when testing for antibodies to other flaviviruses.

Conclusion: Based on these data, multiplex NS1-based protein microarray is a promising tool for surveillance and diagnosis of flaviviruses.

Show MeSH

Related in: MedlinePlus

IgG fluorescent intensity (measured at 647nm) to flaviviruses in serum samples from clinical patients, persons vaccinated with YFV, JEV or TBEV and healthy blood donors in a 1:20 serum dilution.NS1 proteins were spotted in a 0,5mg/ml concentration. Y axis represents the fluorescent intensity. The median signal is depicted as a line. The dotted line represents the calculated cut-off by the ROC used to determine antibody titers.
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pntd.0003580.g002: IgG fluorescent intensity (measured at 647nm) to flaviviruses in serum samples from clinical patients, persons vaccinated with YFV, JEV or TBEV and healthy blood donors in a 1:20 serum dilution.NS1 proteins were spotted in a 0,5mg/ml concentration. Y axis represents the fluorescent intensity. The median signal is depicted as a line. The dotted line represents the calculated cut-off by the ROC used to determine antibody titers.

Mentions: The mean antigen reactivity by NS1 proteins in 1:10 to 1:80 start dilutions was high in homologous DENV, WNV, JEV, SLEV, YFV and Usutu virus (USUV) positive control sera and low in negative control sera and in sera from individuals vaccinated for JEV, TBEV or YFV (p<0.01) with the exception of YFV NS1 antigen with YFV vaccinees (Fig. 2, Fig. 3, and Table 2). Only some NS1 reactivity was observed in samples from blood donors for other antigens (1%).


Spot the difference-development of a syndrome based protein microarray for specific serological detection of multiple flavivirus infections in travelers.

Cleton NB, Godeke GJ, Reimerink J, Beersma MF, Doorn HR, Franco L, Goeijenbier M, Jimenez-Clavero MA, Johnson BW, Niedrig M, Papa A, Sambri V, Tami A, Velasco-Salas ZI, Koopmans MP, Reusken CB - PLoS Negl Trop Dis (2015)

IgG fluorescent intensity (measured at 647nm) to flaviviruses in serum samples from clinical patients, persons vaccinated with YFV, JEV or TBEV and healthy blood donors in a 1:20 serum dilution.NS1 proteins were spotted in a 0,5mg/ml concentration. Y axis represents the fluorescent intensity. The median signal is depicted as a line. The dotted line represents the calculated cut-off by the ROC used to determine antibody titers.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4359159&req=5

pntd.0003580.g002: IgG fluorescent intensity (measured at 647nm) to flaviviruses in serum samples from clinical patients, persons vaccinated with YFV, JEV or TBEV and healthy blood donors in a 1:20 serum dilution.NS1 proteins were spotted in a 0,5mg/ml concentration. Y axis represents the fluorescent intensity. The median signal is depicted as a line. The dotted line represents the calculated cut-off by the ROC used to determine antibody titers.
Mentions: The mean antigen reactivity by NS1 proteins in 1:10 to 1:80 start dilutions was high in homologous DENV, WNV, JEV, SLEV, YFV and Usutu virus (USUV) positive control sera and low in negative control sera and in sera from individuals vaccinated for JEV, TBEV or YFV (p<0.01) with the exception of YFV NS1 antigen with YFV vaccinees (Fig. 2, Fig. 3, and Table 2). Only some NS1 reactivity was observed in samples from blood donors for other antigens (1%).

Bottom Line: Sera from clinical flavivirus patients were used for primary development of the protein microarray.Results show a high IgG and IgM sensitivity and specificity for individual NS1 antigens, and limited cross reactivity, even within serocomplexes.Based on these data, multiplex NS1-based protein microarray is a promising tool for surveillance and diagnosis of flaviviruses.

View Article: PubMed Central - PubMed

Affiliation: Erasmus Medical Centre, Viroscience Department, Rotterdam, The Netherlands; National Institute for Public Health and Environment, Center for Infectious Diseases Research and Screening, Bilthoven, The Netherlands.

ABSTRACT

Background: The family Flaviviridae, genus Flavivirus, holds many of the world's most prevalent arboviral diseases that are also considered the most important travel related arboviral infections. In most cases, flavivirus diagnosis in travelers is primarily based on serology as viremia is often low and typically has already been reduced to undetectable levels when symptoms set in and patients seek medical attention. Serological differentiation between flaviviruses and the false-positive results caused by vaccination and cross-reactivity among the different species, are problematic for surveillance and diagnostics of flaviviruses. Their partially overlapping geographic distribution and symptoms, combined with increase in travel, and preexisting antibodies due to flavivirus vaccinations, expand the need for rapid and reliable multiplex diagnostic tests to supplement currently used methods.

Goal: We describe the development of a multiplex serological protein microarray using recombinant NS1 proteins for detection of medically important viruses within the genus Flavivirus. Sera from clinical flavivirus patients were used for primary development of the protein microarray.

Results: Results show a high IgG and IgM sensitivity and specificity for individual NS1 antigens, and limited cross reactivity, even within serocomplexes. In addition, the serology based on this array allows for discrimination between infection and vaccination response for JEV vaccine, and no cross-reactivity with TBEV and YFV vaccine induced antibodies when testing for antibodies to other flaviviruses.

Conclusion: Based on these data, multiplex NS1-based protein microarray is a promising tool for surveillance and diagnosis of flaviviruses.

Show MeSH
Related in: MedlinePlus