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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

Analysis of cross-reactivity for microarray based flavivirus serology.To visualize the cross-reactivity seen in individual serum samples the maximum calculated titer per sample was set at 100% and all other signals were expressed as a percentage of the highest titer (0–100% on y-axis).
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pntd.0003580.g005: Analysis of cross-reactivity for microarray based flavivirus serology.To visualize the cross-reactivity seen in individual serum samples the maximum calculated titer per sample was set at 100% and all other signals were expressed as a percentage of the highest titer (0–100% on y-axis).

Mentions: In order to study cross-reactivity within and between serocomplexes, serum samples were serially diluted and titers were calculated in R. Typical individual patient profiles are shown in Fig. 4. To quantify the degree of cross-reactivity, the ratio of the signal for each antigen to the maximum signal measured for that serum (typically the homologous antigen) was calculated (Fig. 5). With one exception for IgG (serum sample #4), all patients had the highest IgG and IgM reactivity with the homologous NS1 antigen. High level IgG reactivity to a second antigen was observed for two of the DENV patients (against WNV and JEV, respectively) and for 2 JEV patients (against DENV) (Fig. 5a). One serum sample from a JEV patient (serum sample 4) had a higher titer DENV NS1 in comparison to JEV NS1. For IgM, only homologous reactivity was observed.


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)

Analysis of cross-reactivity for microarray based flavivirus serology.To visualize the cross-reactivity seen in individual serum samples the maximum calculated titer per sample was set at 100% and all other signals were expressed as a percentage of the highest titer (0–100% on y-axis).
© Copyright Policy
Related In: Results  -  Collection

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

pntd.0003580.g005: Analysis of cross-reactivity for microarray based flavivirus serology.To visualize the cross-reactivity seen in individual serum samples the maximum calculated titer per sample was set at 100% and all other signals were expressed as a percentage of the highest titer (0–100% on y-axis).
Mentions: In order to study cross-reactivity within and between serocomplexes, serum samples were serially diluted and titers were calculated in R. Typical individual patient profiles are shown in Fig. 4. To quantify the degree of cross-reactivity, the ratio of the signal for each antigen to the maximum signal measured for that serum (typically the homologous antigen) was calculated (Fig. 5). With one exception for IgG (serum sample #4), all patients had the highest IgG and IgM reactivity with the homologous NS1 antigen. High level IgG reactivity to a second antigen was observed for two of the DENV patients (against WNV and JEV, respectively) and for 2 JEV patients (against DENV) (Fig. 5a). One serum sample from a JEV patient (serum sample 4) had a higher titer DENV NS1 in comparison to JEV NS1. For IgM, only homologous reactivity was observed.

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