Retrospective Study
Copyright ©The Author(s) 2021. Published by Baishideng Publishing Group Inc. All rights reserved.
World J Methodol. Nov 20, 2021; 11(6): 294-301
Published online Nov 20, 2021. doi: 10.5662/wjm.v11.i6.294
Comparison of indirect immunofluorescence and western blot method in the diagnosis of hantavirus infections
Tatjana Vilibic-Cavlek, Ljubo Barbic, Vladimir Stevanovic, Vladimir Savic, Anna Mrzljak, Maja Bogdanic, Irena Tabain
Tatjana Vilibic-Cavlek, Anna Mrzljak, School of Medicine, University of Zagreb, Zagreb 10000, Croatia
Tatjana Vilibic-Cavlek, Maja Bogdanic, Irena Tabain, Department of Virology, Croatian Institute of Public Health, Zagreb 10000, Croatia
Ljubo Barbic, Vladimir Stevanovic, Department of Microbiology and Infectious Diseases with Clinic, Faculty of Veterinary Medicine, University of Zagreb, Zagreb 10000, Croatia
Vladimir Savic, Poultry Centre, Croatian Veterinary Institute, Zagreb 10000, Croatia
Anna Mrzljak, Department of Gastroenterology and Hepatology, University Hospital Zagreb, Zagreb 10000, Croatia
Author contributions: Vilibic-Cavlek T made contributions to the conception and design of the study and was involved in drafting and revising the manuscript; Stevanovic V, Savic V, Mrzljak A, and Bogdanic M were involved in collecting data and drafting and writing the manuscript; Barbic L and Tabain I revised the manuscript critically; All authors read and approved the final manuscript.
Institutional review board statement: The study was approved by the Ethics Committee of the Croatian Institute of Public Health, No. 030-02/17-10/1.
Informed consent statement: All study participants or their legal guardian provided informed written consent about personal and medical data collection prior to study enrolment.
Conflict-of-interest statement: All authors declare no conflict of interest.
Data sharing statement: No sharing data.
Open-Access: This article is an open-access article that was selected by an in-house editor and fully peer-reviewed by external reviewers. It is distributed in accordance with the Creative Commons Attribution NonCommercial (CC BY-NC 4.0) license, which permits others to distribute, remix, adapt, build upon this work non-commercially, and license their derivative works on different terms, provided the original work is properly cited and the use is non-commercial. See: http://creativecommons.org/Licenses/by-nc/4.0/
Corresponding author: Tatjana Vilibic-Cavlek, MD, PhD, Associate Professor, Department of Virology, Croatian Institute of Public Health, Rockefellerova 12, Zagreb 10000, Croatia. tatjana.vilibic-cavlek@hzjz.hr
Received: January 2, 2021
Peer-review started: January 2, 2021
First decision: July 8, 2021
Revised: July 16, 2021
Accepted: August 31, 2021
Article in press: August 31, 2021
Published online: November 20, 2021
Processing time: 316 Days and 15.6 Hours
Abstract
BACKGROUND

Serologic cross-reactivity between hantaviruses often complicates the interpretation of the results.

AIM

To analyze the diagnostic value of indirect immunofluorescence assay (IFA) and western blot (WB) in the diagnosis of hantavirus infections.

METHODS

One hundred eighty-eight serum samples from Puumala (PUUV) and Dobrava (DOBV) orthohantavirus infected patients were analyzed. Serology was performed using commercial tests (Euroimmun, Lübeck, Germany).

RESULTS

Using IFA, 49.5% of acute-phase samples showed a monotypic response to PUUV, while 50.5% cross-reacted with other hantaviruses. The overall cross-reactivity was higher for immunoglobulin G (IgG) (50.0%) than for immunoglobulin M (IgM) (25.5%). PUUV IgM/IgG antibodies showed low/moderate reactivity with orthohantaviruses Hantaan (12.3%/31.5%), Seoul (7.5%/17.8%), DOBV (5.4%/ 28.1%), and Saaremaa (4.8%/15.7%). Both DOBV IgM and IgG antibodies were broadly reactive with Hantaan (76.2%/95.2%), Saaremaa (80.9%/83.3%), and Seoul (78.6%/85.7%) and moderate with PUUV (28.5%/38.1%). Using a WB, serotyping was successful in most cross-reactive samples (89.5%).

CONCLUSION

The presented results indicate that WB is more specific than IFA in the diagnosis of hantavirus infections, confirming serotype in most IFA cross-reactive samples.

Keywords: Hantaviruses; Serology; Cross-reactivity; Indirect immunofluorescence; Western blot

Core Tip: Serologic cross-reactivity among hantaviruses often complicates the interpretation of the results. The overall cross-reactivity is generally higher for immunoglobulin G antibodies than for immunoglobulin M antibodies. Western blot seems to be a more specific serology method than indirect immunofluorescence assay in the diagnosis of hantavirus infections, confirming serotype in the majority of cross-reactive samples detected by indirect immunofluorescence assay.