Detection of Chikungunya Virus Antigen by a Novel Rapid Immunochromatographic Test

Abstract

Chikungunya fever is a mosquito-borne disease of key public health importance in tropical and subtropical countries. Although severe joint pain is the most distinguishing feature of chikungunya fever, diagnosis remains difficult because the symptoms of chikungunya fever are shared by many pathogens, including dengue fever. The present study aimed to develop a new immunochromatographic diagnosis test for the detection of chikungunya virus antigen in serum. Mice were immunized with isolates from patients with Thai chikungunya fever, East/Central/South African genotype, to produce mouse monoclonal antibodies against chikungunya virus. Using these monoclonal antibodies, a new diagnostic test was developed and evaluated for the detection of chikungunya virus. The newly developed diagnostic test reacted with not only the East/Central/South African genotype but also with the Asian and West African genotypes of chikungunya virus. Testing of sera from patients suspected to have chikungunya fever in Thailand (n ‫؍‬ 50), Laos (n ‫؍‬ 54), Indonesia (n ‫؍‬ 2), and Senegal (n ‫؍‬ 6) revealed sensitivity, specificity, and real-time PCR (RT-PCR) agreement values of 89.4%, 94.4%, and 91.1%, respectively. In our study using serial samples, a new diagnostic test showed high agreement with the RT-PCR within the first 5 days after onset. A rapid diagnostic test was developed using mouse monoclonal antibodies that react with chikungunya virus envelope proteins. The diagnostic accuracy of our test is clinically acceptable for chikungunya fever in the acute phase.

Diagnostic potential of monoclonal antibodies against the capsid protein of chikungunya virus for detection of recent infection

“…We explored the use of anti-C MAbs in IgM capture ELISA for the detection of recent CHIKV infection. The E protein or the E protein gene of CHIKV has always been considered a major target for the diagnosis of CHIKV infection in serological and molecular assays [39, 41,42,51,52], but recently, the C protein has also been explored as a target for CHIKV diagnosis in various assays. Reports indicate 85 % sensitivity and 100 % specificity [39] and 97 % accordance of indirect IgM detection ELISA using recombinant C protein when compared to a commercial kit [40].…”

“…MAbs to E2 protein successfully detected CHIKV derived from mosquito and human cells in antigen-capture ELISA [41]. However, Okabayashi et al correlated higher sensitivity of the E1 MAb-based ICA test compared to RT-PCR within the early phase of infection with the degree to which RNA and antigen were preserved in serum samples [42].…”

Recent progress on chikungunya virus research

“…However, only a few commercial CHIKV antigen detection kits using Mab are available, and they have different level of sensitivity (30%-90%) towards different viral genotypes (Okabayashi et al, 2015;Huits et al, 2017). Although some antigen detection methods developed in laboratories have higher sensitivities, their specificities are not clearly defined, as the employment of polyclonal antibodies purified from patients or those from animals immunized with whole virus made standardization difficult.…”

Diagnostic tools for tackling febrile illness and enhancing patient management

“…The net result is that health workers, especially those working in private clinics, and even patients going to pharmacies, have access to POC tests that have in fact very low sensitivity and/or specificity. For example, LFT for chikungunya is commercialized, although it has a sensitivity of maximum 50% in the field [190][191][192].…”