Indirect immunofluorescence assay for the simultaneous detection of antibodies against clinically important old and new world hantaviruses
Authors: Jonas Schmidt-Chanasit
Paul K S Chan
Paula Padula
Merle Hanke
Cornelia Ceianu
Sabine Lederer
Olli Vapalahti
Pablo A Vial
Daryl Dick
Winfried Stoecker
Heinz Feldmann
Antti Vaheri
Karen Sonnenberg
Matthias Niedrig
Raluca Panculescu-Gatej
Erik Lattwein
Tatjana Av≈°iƒç-≈Ωupanc
√Öke Lundkvist
Paul Heyman
Journal: PLoS neglected tropical diseases (2013)
Discipline: IMMUNOLOGY
Keywords: hantaviruses; multiparametric indirect immunofluorescence assays; serum antibodies; biochip mosaics; IgG
Affiliations: EUROIMMUN Medizinische Labordiagnostika AG, Luebeck, Germany.

Abstract:  In order to detect serum antibodies against clinically important Old and New World hantaviruses simultaneously, multiparametric indirect immunofluorescence assays (IFAs) based on biochip mosaics were developed. Each of the mosaic substrates consisted of cells infected with one of the virus types Hantaan (HTNV), Puumala (PUUV), Seoul (SEOV), Saaremaa (SAAV), Dobrava (DOBV), Sin Nombre (SNV) or Andes (ANDV). For assay evaluation, serum IgG and IgM antibodies were analyzed using 184 laboratory-confirmed hantavirus-positive sera collected at six diagnostic centers from patients actively or previously infected with the following hantavirus serotypes: PUUV (Finland, n=97); SEOV (China, n=5); DOBV (Romania, n=7); SNV (Canada, n=23); ANDV (Argentina and Chile, n=52). The control panel comprised 89 sera from healthy blood donors. According to the reference tests, all 184 patient samples were seropositive for hantavirus-specific IgG (n=177; 96%) and/or IgM (n=131; 72%), while all control samples were tested negative. In the multiparametric IFA applied in this study, 183 (99%) of the patient sera were IgG and 131 (71%) IgM positive (accordance with the reference tests: IgG, 96%; IgM, 93%). Overall IFA sensitivity for combined IgG and IgM analysis amounted to 100% for all serotypes, except for SNV (96%). Of the 89 control sera, 2 (2%) showed IgG reactivity against the HTNV substrate, but not against any other hantavirus. Due to the high cross-reactivity of hantaviral nucleocapsid proteins, endpoint titrations were conducted, allowing serotype determination in &gt;90% of PUUV- and ANDV-infected patients. Thus, multiparametric IFA enables highly sensitive and specific serological diagnosis of hantavirus infections and can be used to differentiate PUUV and ANDV infection from infections with Murinae-borne hantaviruses (e.g. DOBV and SEOV). 

Retracted: No

PubPeer comments:
 Figure 1. The two slides shown her appear to show a very similar background, yet with a different set of center mosaics.  In addition, both slides, but in particular the bottom slide, shows a remarkable similarity to the Phospholipase A2 slide shown in Figure 1 of Schlumberger et al.  Differential diagnosis of membranous nephropathy with autoantibodies to phospholipase A2 receptor 1 , Autoimmunity Reviews (2014), doi: 10.1016 j.autrev.2013.09.005. Thirdly, all three slides appear to show the same repetitive patterns. I have marked these already in the Schlumberger photo.  Could the authors please clarify if these are meant to be photocomposites of a future product, or if these are representing a real product? , 
 The same product slide might be visible in another paper from partially the same authors. Here is Figure 1 from Niedrig et al., Clinical and vaccine immunology : CVI (2008), DOI: 10.1128 CVI.00078-07. In the Niedrig 2008 paper, the slide is designed to detect Yellow Fever Virus.  , 
 The six-pack Hantavirus 1 chip (top photo in Figure 1 of this paper) could easily be confused with the Granulocyte mosaic chip from Euroimmun, because they look very similar. That chip could be found in Figure 1 of Gianna Mastroianni-Kirsztajn, Nora Hornig and Wolfgang Schlumberger Autoantibodies in renal diseases u2013 clinical significance and recent developments in serological detectiont Frontiers in Immunology (2015) DOI: 10.3389 fimmu.2015.00221 We can distinguish them, though, because they have a different assay name printed on them.  , 
 November 9, 2020 Notice of Republication (but labeled as Correction ) found here: https: journals.plos.org plosntds article?id=10.1371 journal.pntd.0008864  Following publication of this article [1], concerns were raised about regions of similarity between the images of the indirect immunofluorescence assay (IFA) microscope slides for detection of hantavirus-specific antibodies shown in Fig 1 and images of IFA slides for detection of different antibodies published in other articles [2u20134]. The authors clarify that the images in Fig 1 were digitally created using image editing software to combine images of a slide background, reaction fields and assay labels. The images in Fig 1 are for illustrative purposes only, to support understanding of slide configuration and methodology. Fig 1 does not contribute to the results data or scientific conclusions of the study. In light of concerns that the originally published Fig 1 includes previously published image elements that are not licensed for reproduction and distribution under the terms of the CC-BY license, the original article was republished on October 22, 2020, with a revised Fig 1 in which slide configuration is shown as a schematic diagram. Please download this article again to view the correct version. The revised Fig 1 is provided here for reference. Here is the new Figure 1:  ,