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. 2022 Mar;24(2):104890.
doi: 10.1016/j.micinf.2021.104890. Epub 2021 Oct 7.

Developing and validating a modified enzyme linked immunosorbent assay method for detecting HEV IgG antibody from dried blood spot (DBS) samples in endemic settings

Rosy Sultana  1 , Taufiqur Rahman Bhuiyan  2 , Afsana Shirin Sathi  2 , Salma Sharmin  2 , Sharmina Yeasmin  3 , Muhammad Ikhtear Uddin  2 , Md Saruar Bhuiyan  2 , Kaiissar Mannoor  4 , Muhammad Manjurul Karim  5 , K Zaman  2 , Firdausi Qadri  6
Affiliations

Developing and validating a modified enzyme linked immunosorbent assay method for detecting HEV IgG antibody from dried blood spot (DBS) samples in endemic settings

Rosy Sultana et al. Microbes Infect. 2022 Mar.

Abstract

Serological analysis is an integral part of laboratory practice nowadays. The present study was aimed to develop and validate a modified Enzyme linked Immunosorbent Assay (ELISA) for determination of IgG antibody against Hepatitis E Virus (HEV) using dried blood spots (DBS) and corresponding plasma samples. A total of 65 samples (45 HEV patients, 20 healthy controls) were analyzed. DBS and plasma samples demonstrated equivalent optical densities for detecting anti-HEV IgG. A highly significant correlation was observed between plasma and DBS sample absorbances (R2 = 0.98; p < 0.001) at dilution 1:200, indicating true agreement between the two procedures. The assay exhibited decent linearity and showed no effect of physiological hematocrit on assay performance. Data suggested recommendable promise in using DBS as a suitable alternative to plasma samples to determine HEV IgG antibody evidenced by significant correlation with plasma results. Therefore, identical method for processing DBS specimens including it's proper storage is recommended for implementation of a modified ELISA in different settings.

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Conflict of interest statement

Declaration of competing interest All authors: No reported conflicts. All authors have submitted the ICMJE Form for Disclosure of Potential Conflicts of Interest. Conflicts that the editors consider relevant to the content of the manuscript have been disclosed.

Figures

Figure 1:
Figure 1:
Stratification of HEV IgG positive and negative samples of the hepatitis cases and healthy controls. Y axis shows the numbers. Numbers over the bars indicate number of subjects in each group.
Figure 2:
Figure 2:
Comparison of optical densities for measurement of anti-HEV IgG antibody in DBS and corresponding plasma of the patients with HEV and healthy control subjects. Y axis showed the mean absorbance (optical densities) at 450 nm with SEM.
Figure 3:
Figure 3:
Correlation between DBS and corresponding plasma samples for determination of HEV IgG antibody of the study subjects. Optical densities (OD) of DBS are shown in X-axis and optical densities (OD) of corresponding plasma are shown in Y-axis that were measured at 450 nm.
See this image and copyright information in PMC

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