The Comparison of Sensitivity and Specificity of ELISA-based Microneutralization Test with Hemagglutination Inhibition Test to Evaluate Neutralizing Antibody against Influenza Virus (H1N1)

Ahmad Tavakoli ,Fatemeh Rezaei ,Zahra Noroozbabaei ,Gazal Sadat Fatemi Nasab ,Fatemeh Adjaminezhad-Fard ,Talat Mokhtari‐Azad

doi:10.7176/jmpb/65-08

Abstract

The most common serological assay to measure anti-influenza antibodies is hemagglutination inhibition (HI) assay. Recently, neutralizing antibodies against influenza virus infection or vaccination can also be detected using microneutralization assays and occasionally, have greater sensitivity than the standard HI assays. The study aimed to compare the sensitivity and specificity of ELISA-based microneutralization (microNT-ELISA) and conventional HI assays in order to detect influenza H1N1 virus antibodies.MicroNT-ELISA was set up according to the WHO Manual on Influenza Diagnosis and Surveillance in Virology Department of Tehran University of Medical Sciences for the detection of neutralizing antibodies against H1N1 influenza virus in 2013. Fifty serum samples were analyzed with both HI and microNT-ELISA assays. Correlation between methods was calculated by linear regression analysis.The linear correlation coefficient squares, R2, of microNT-ELISA and HI test was 0.61 (P<0.0001) and we observed a high index of coincidence between the two tests. According to McNemar's test, there was no statistically significant difference between these two assays (P>0.05).The sensitivity and specificity of microNT-ELISA assay were high (87% and 73%, respectively) and closely related to gold standard test results. Therefore, microNT-ELISA is recommended as an alternative or complementary test to conventional HI assay for serological and epidemiological purposes.

Highlights

Seasonal pandemics and epidemics of influenza result in considerable number of mortalities and morbidities and it is regarded as a major health issue in the entire world 1
Serum sample was obtained from 188 patients, of which 101 patients had recently suffered from influenza
Among the healthy persons in our study 8 showed positive result and 79 showed negative result when microneutralization test was applied to assess the serological response of neutralizing antibodies in patients with influenza A

Summary

Introduction

Seasonal pandemics and epidemics of influenza result in considerable number of mortalities and morbidities and it is regarded as a major health issue in the entire world 1. It can be detected with several types of techniques 2. Particular antibodies against certain viral antigen as humoral immune response, are produced as result of infection by the virus or if vaccination of the virus. These antibodies are detectable for duration of about two to three weeks after the emergence of the symptoms with the help of serological testing 5. The advantages of serological assays like, Hemagglutination inhibition test and ELISA based Microneutralization test, are that these are diagnostically helpful and that they provide essential knowledge when use for immunological and epidemiological surveys and in evaluation of immunogenicity of a vaccine 7

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