Development of a neutralization assay based on the pseudotyped chikungunya virus of a Korean isolate

Chung, Woo Chang; Hwang, Kwang Yeon; Kang, Sang-Won; Kim, Jae Ouk; Song, Mee Hyun

doi:10.1007/s12275-020-9384-0

Cited by 6 publications

(8 citation statements)

References 38 publications

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“…This might be confirmed by the detection of the presence of IgG antibodies in patients [ 28 , 29 ]. PRNT is used to measure the titer of neutralizing antibodies in the serum by counting the number of plaques of viral infection [ 33 ]. This test has been used conventionally in laboratories for vaccine efficacy that requires the hazardous live CHIKV, and requires fewer days to complete when compared to the RDT [ 33 , 34 ].…”

Section: Discussionmentioning

confidence: 99%

“…PRNT is used to measure the titer of neutralizing antibodies in the serum by counting the number of plaques of viral infection [ 33 ]. This test has been used conventionally in laboratories for vaccine efficacy that requires the hazardous live CHIKV, and requires fewer days to complete when compared to the RDT [ 33 , 34 ]. Currently, neutralization assays, such as the non-infectious virus replicon particles and pseudotyped viruses, have been developed [ 35 , 36 ].…”

Section: Discussionmentioning

confidence: 99%

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Prevalence of Malaria and Chikungunya Co-Infection in Febrile Patients: A Systematic Review and Meta-Analysis

et al. 2021

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Co-infection with malaria and chikungunya (CHIKV) could exert a significant public health impact with infection misdiagnosis. Therefore, this study aimed to collect qualitative and quantitative evidence of malaria and CHIKV co-infection among febrile patients. Methods: Potentially relevant studies were identified using PubMed, Web of Science, and Scopus. The bias risk of the included studies was assessed using the checklist for analytical cross-sectional studies developed by the Joanna Briggs Institute. The pooled prevalence of malaria and CHIKV co-infection among febrile patients and the pooled prevalence of CHIKV infection among malaria patients were estimated with the random effect model. The odds of malaria and CHIKV co-infection among febrile patients were also estimated using a random effect model that presumed the heterogeneity of the outcomes of the included studies. The heterogeneity among the included studies was assessed using the Cochran Q test and I2 statistics. Publication bias was assessed using the funnel plot and Egger’s test. Results: Of the 1924 studies that were identified from the three databases, 10 fulfilled the eligibility criteria and were included in our study. The pooled prevalence of malaria and CHIKV co-infection (182 cases) among febrile patients (16,787 cases), stratified by diagnostic tests for CHIKV, was 10% (95% confidence interval (CI): 8–11%, I2: 99.5%) using RDT (IgM), 7% (95% CI: 4–10%) using the plaque reduction neutralization test (PRNT), 1% (95% CI: 0–2%, I2: 41.5%) using IgM and IgG ELISA, and 4% (95% CI: 2–6%) using real-time RT-PCR. When the prevalence was stratified by country, the prevalence of co-infection was 7% (95% CI: 5–10%, I2: 99.5%) in Nigeria, 1% (95% CI: 0–2%, I2: 99.5%) in Tanzania, 10% (95% CI: 8–11%) in Sierra Leone, 1% (95% CI: 0–4%) in Mozambique, and 4% (95% CI: 2–6%) in Kenya. The pooled prevalence of CHIKV infection (182 cases) among malaria patients (8317 cases), stratified by diagnostic tests for CHIKV, was 39% (95% CI: 34–44%, I2: 99.7%) using RDT (IgM), 43% (95% CI: 30–57%) using PRNT, 5% (95% CI: 3–7%, I2: 5.18%) using IgM and IgG ELISA, and 9% (95% CI: 6–15%) using real-time RT-PCR. The meta-analysis showed that malaria and CHIKV co-infection occurred by chance (p: 0.59, OR: 0.32, 95% CI: 0.6–1.07, I2: 78.5%). Conclusions: The prevalence of malaria and CHIKV co-infection varied from 0% to 10% as per the diagnostic test for CHIKV infection or the country where the co-infection was reported. Hence, the clinicians who diagnose patients with malaria infections in areas where two diseases are endemic should further investigate for CHIKV co-infection to prevent misdiagnosis or delayed treatment of concurrent infection.

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Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Prevalence of Malaria and Chikungunya Co-Infection in Febrile Patients: A Systematic Review and Meta-Analysis

et al. 2021

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“…However, measurement of luciferase activity is complicate and expensive. Dual reporters (ZsGreen1 and luciferase) have been increasingly engineered into PsV of this study and other reports 27 , 28 . The fluorescent reporter ZsGreen1 in PsV can be used to monitor efficiency of transfection and titration, which is more convenient and cheaper than the reporter luciferase.…”

Section: Discussionmentioning

confidence: 78%

“…In addition, visualization and counting of plaques in PRNT are time-consuming and laborious procedures. Many studies have demonstrated that the PsV neutralization test could be a good substitute for the plaque reduction test 27 , 34 . In order to overcome the limitations of PRNT mentioned above, here we developed a PsV neutralization assay.…”

Section: Discussionmentioning

confidence: 99%

“…In order to overcome the limitations of PRNT mentioned above, here we developed a PsV neutralization assay. In this assay, luciferase activity was used to quantify the neutralization activity, which had been considered to be semi-automatic, high throughput, sensitive and accurate 27 , 34 , 36 . As compared with the neutralization activity of VSV-G PsV, our result showed that neutralization assay of CHIKV PsV is dose dependent and CHIKV specific (Fig.…”

Section: Discussionmentioning

confidence: 99%

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Preparation and application of chikungunya pseudovirus containing double reporter genes

Ding

et al. 2022

Sci Rep

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Chikungunya virus (CHIKV), a highly infectious and rapidly spread viral pathogen, is classified as a pathogenic agent at the biosafety level 3. Operation of live authentic CHIKV needs a specific laboratory with the P3 or above containment, which greatly confines the CHIKV-associated studies. To establish an evaluation system of CHIKV that can be utilized in a BSL2 laboratory, we constructed a pseudovirus (PsV) system of CHIKV containing double reporter genes (ZsGreen1 and luciferase). The fluorescent ZsGreen1 is a convenient and cheap reporter for monitoring the efficiency of transfection and titration of PsV. The enzyme luciferase is a sensitive reporter for the application of PsV to neutralization assay or drug screening. The CHIKV PsV produced in this study, with a titer of up to 3.16 × 106 TU/ml, was confirmed by Western blotting and transmission electronic microscopy (TEM). Finally, we developed a microneutralization assay with the CHIKV PsV produced in this study, which was successfully applied to evaluate neutralizing activities of convalescent sera from CHIKV-infected patients. In summary, we have established a convenient and sensitive double-reporter CHIKV pseudovirus system, which provides a safe and effective platform for screening anti-CHIKV drugs and evaluating vaccines against CHIKV.

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Pseudotyped Viruses for the Alphavirus Chikungunya Virus

Huang

Wang

2023

Advances in Experimental Medicine and Biology

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Development of a neutralization assay based on the pseudotyped chikungunya virus of a Korean isolate

Cited by 6 publications

References 38 publications

Prevalence of Malaria and Chikungunya Co-Infection in Febrile Patients: A Systematic Review and Meta-Analysis

Prevalence of Malaria and Chikungunya Co-Infection in Febrile Patients: A Systematic Review and Meta-Analysis

Preparation and application of chikungunya pseudovirus containing double reporter genes

Pseudotyped Viruses for the Alphavirus Chikungunya Virus

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