Under certain rare circumstances, PRNT testing can also be affected by cross reactivity between flaviviruses. These requirements generate long turnaround times that can reach 5 days before obtaining confirmations. In order to bolster the specificity of results, plaque reduction neutralization test (PRNT) has been introduced to confirm positive ELISA results however, this method is currently difficult to implement in low and middle income countries (LMICs) as it requires manipulations of the live virus in cell culture, and therefore, can only be performed in specialized biosafety level 3 (BS元) containment laboratories by highly trained personnel. It has been proven that vaccination or past infections with other flaviviruses considerably increases the level of cross reactivity, rendering interpretation of serological results very difficult, especially in flavivirus endemic regions. However, the biggest challenge of serological assays is the cross-reactivity of ZIKV antibodies with other flaviviruses, especially with dengue virus. This method should, therefore, be extended to serological diagnostic techniques for other members of the flavivirus genus and for use in IgM diagnostic testing.ĭuring ZIKV infection, viremia generally lasts less than 7 days and direct virus detection can be accomplished using reverse transcription-polymerase chain reaction (RT-PCR) or virus isolation during the acute phase however, as viremia is fleeting and resolves usually within the first week of illness, current laboratory diagnostics for ZIKV are mainly based on indirect methods aiming to detect IgM and IgG class antibodies. These results show the usefulness of the recombinant envelope domain III as an alternative to standard whole virus proteins for ZIKV diagnostics as it improves the sensitivity and specificity of IgG ELISA assay when used as an immunogen. Using a total of 367 clinical samples, we showed that the EDIII-ELISA was able to detect IgG antibodies against ZIKV with high sensitivity of 100.0% and specificity of 94.7% when compared to plaque reduction neutralization tests (PRNTs) as the gold standard and using 0.208 as the cut-off OD value. Here, we describe an indirect ELISA to detect specific IgG antibodies using the ZIKV envelope domain III (EDIII) protein expressed in Drosophila S2 cells as an immunogen. However, current serology tests using whole virus antigens frequently suffer from cross reactivity issues, delays, and technical complexity, especially in low and middle income countries (LMICs) and endemic countries. Since the viremia during ZIKV infection is fleeting, serological testing is highly valuable to inform diagnosis. Zika virus (ZIKV) diagnostics are crucial for proper antenatal and postnatal care and also for surveillance and serosurvey studies.
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