Please use this identifier to cite or link to this item: https://doi.org/10.1371/journal.pntd.0000753
Title: Evaluation of chikungunya diagnostic assays: Differences in sensitivity of serology assays in two independent outbreaks
Authors: Yap G.
Pok K.-Y.
Lai Y.-L.
Hapuarachchi H.-C.
Chow A. 
Leo Y.-S. 
Tan L.-K.
Ng L.-C.
Keywords: immunoglobulin M
diagnostic agent
immunoglobulin M
virus antibody
virus antigen
virus envelope protein
virus RNA
antigen detection
article
chikungunya
Chikungunya alphavirus
clinical article
controlled study
enzyme linked immunosorbent assay
human
intermethod comparison
molecular probe
nonhuman
real time polymerase chain reaction
reverse transcription polymerase chain reaction
sensitivity analysis
sensitivity and specificity
serology
Alphavirus infection
amino acid substitution
blood
Chikungunya alphavirus
comparative study
epidemic
evaluation
genetics
immunoassay
immunology
methodology
point mutation
polymerase chain reaction
Singapore
virology
Alphavirus Infections
Amino Acid Substitution
Antibodies, Viral
Antigens, Viral
Chikungunya virus
Disease Outbreaks
Humans
Immunoassay
Immunoglobulin M
Point Mutation
Polymerase Chain Reaction
RNA, Viral
Sensitivity and Specificity
Singapore
Viral Envelope Proteins
Virology
Issue Date: 2010
Citation: Yap G., Pok K.-Y., Lai Y.-L., Hapuarachchi H.-C., Chow A., Leo Y.-S., Tan L.-K., Ng L.-C. (2010). Evaluation of chikungunya diagnostic assays: Differences in sensitivity of serology assays in two independent outbreaks. PLoS Neglected Tropical Diseases 4 (7) : e753. ScholarBank@NUS Repository. https://doi.org/10.1371/journal.pntd.0000753
Rights: Attribution 4.0 International
Abstract: Background: The sensitivity and specificity of two in-house MAC-ELISA assays were tested and compared with the performance of commercially-available CTK lateral flow rapid test and EUROIMMUN IFA assays for the detection of anti- Chikungunya virus (CHIKV) IgM. Each MAC-ELISA assay used a whole virus-based antigen derived from genetically distinct CHIKV strains involved in two chikungunya disease outbreaks in Singapore (2008); a January outbreak strain with alanine at amino acid residue 226 of the E1 glycoprotein (CHIKV-A226) and a May-to-September outbreak strain that possessed valine at the same residue (CHIKV-226V). We report differences in IgM detection efficacy of different assays between the two outbreaks. The sensitivities of two PCR protocols were also tested. Methods and Findings: For sera from January outbreak, the average detection threshold of CTK lateral flow test, MACELISAs and EUROIMMUN IFA assays was 3.75, 4.38 and 4.88 days post fever onset respectively. In contrast, IgM detection using CTK lateral flow test was delayed to more than 7 days after fever onset in the second outbreak sera. However, MACELISA using CHIKV-226V detected IgM in the second outbreak sera 3.96 days after fever onset, which was approximately one day earlier compared to the same assay using CHIKV-A226 (4.86 days). Specificity was 100% for both commercial assays, and 95.6% for the in-house MAC-ELISAs. For sensitivity determination of the PCR protocols, the probe-based real time RT-PCR method was found to be 10 times more sensitive than one based on SYBR Green. Conclusion: Our findings suggested that the two strains of CHIKV using variants A226 and 226V resulted in variation in sensitivities of the assays evaluated. We postulated that the observed difference in antigen efficacy could be due to the amino acid substitution differences in viral E1 and E2 envelope proteins, especially the E1-A226V substitution. This evaluation demonstrates the importance of appraisal of different diagnostic assays before their application in clinical and operational settings. © 2010 Yap et al.
Source Title: PLoS Neglected Tropical Diseases
URI: https://scholarbank.nus.edu.sg/handle/10635/161660
ISSN: 19352727
DOI: 10.1371/journal.pntd.0000753
Rights: Attribution 4.0 International
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