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:
Corresponding author: Tatjana Vilibic-Cavlek, MD, PhD, Associate Professor, Department of Virology, Croatian Institute of Public Health, Rockefellerova 12, Zagreb 10000, Croatia.
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
Research background

The cross-reactivity among hantaviruses often complicates the interpretation of serology results, especially in areas where different hantaviruses co-circulate.

Research motivation

Data on the comparison of different serologic methods in the diagnosis of hantaviruses are scarce.

Research objectives

This study aimed to analyze the diagnostic value of indirect immunofluorescence (IFA) and western blot (WB) methods in diagnosing hantavirus infections.

Research methods

A commercial IFA was used to detect immunoglobulin M (IgM)/immunoglobulin G (IgG) antibodies to the most common orthohantaviruses: Puumala (PUUV), Dobrava (DOBV), Hantaan (HTNV), Seoul (SEOV), and Saaremaa (SAAV). Cross-reactive samples were additionally tested by a commercial WB using PUUV, DOBV, and HTNV antigens.

Research results

Using IFA, 49.5% of acute-phase serum samples reacted only with the homologous PUUV antigen, while in 50.5% samples, cross-reactive IgM and/or IgG antibodies were found. PUUV IgM/IgG antibodies cross-reacted with HTNV (12.3%/31.5%), SEOV (7.5%/17.8%), DOBV (5.4%/28.1%), and SAAV (4.8%/15.7%). Both DOBV IgM and IgG antibodies were broadly reactive with HTNV (76.2%/95.2%), SAAV (80.9%/83.3%), and SEOV (78.6%/85.7%) and moderate with PUUV (28.5%/38.1%). Using a WB, serotyping was successful in 89.5% cross-reactive samples.

Research conclusions

WB seems to be more specific than IFA, confirming hantavirus serotype in the majority of cross-reactive samples detected by IFA.

Research perspectives

Further studies on a large sample caused by different hantavirus serotypes are needed.