Development of Highly Sensitive Sandwich ELISA for the Early-Phase Diagnosis of Chikungunya Virus Utilizing rE2-E1 Protein.
B-cell epitopes
IgG
chikungunya
hamster antibodies
recombinant E2-E1
sandwich ELISA
Journal
Infection and drug resistance
ISSN: 1178-6973
Titre abrégé: Infect Drug Resist
Pays: New Zealand
ID NLM: 101550216
Informations de publication
Date de publication:
2022
2022
Historique:
received:
03
11
2021
accepted:
03
06
2022
entrez:
4
8
2022
pubmed:
5
8
2022
medline:
5
8
2022
Statut:
epublish
Résumé
Chikungunya is caused by an alpha virus transmitted to humans by an infected mosquito. Infection is generally considered to be self-limiting and non-critical. Chikungunya infection may be diagnosed by severe joint pain with fever, but it is difficult to diagnose because the symptoms of chikungunya are common to many pathogens, including dengue fever. Diagnosis mainly depends on viral culture, reverse transcriptase polymerase chain reaction (RT-PCR), and IgM ELISA. Early and accurate diagnosis of the virus can be achieved by the application of PCR methods, but the high cost and the need for a thermal cycler restrict the use of such methods. On the other hand, antibody-based IgM ELISA is considered to be inexpensive, but antibodies against chikungunya virus (CHIKV) only develop after 4 days of infection, so it has limited application in the earlier diagnosis of viral infection and the management of patients. Because of these challenges, a simple antigen-based sensitive, specific, and rapid detection method is required for the early and accurate clinical diagnosis of chikungunya. The amino acid sequence of CHIKV ectodomain E1 and E2 proteins was analyzed using bioinformatics tools to determine the antigenic residues, particularly the B-cell epitopes and their characteristics. Recombinant E2-E1 CHIKV antigen was used for the development of polyclonal antibodies in hamsters and IgG was purified. Serological tests of 96 CHIKV patients were conducted by antigen-capture ELISA using primary antibodies raised against rCHIKV E2-E1 in hamsters and human anti-CHIKV antibodies. We observed high specificity and sensitivity, of 100% and 95.8%, respectively, and these values demonstrate the efficiency of the test as a clinical diagnostic tool. There was no cross-reactivity with samples taken from dengue patients. Our simple and sensitive sandwich ELISA for the early-phase detection of CHIKV infection may be used to improve the diagnosis of chikungunya.
Identifiants
pubmed: 35924014
doi: 10.2147/IDR.S347545
pii: 347545
pmc: PMC9342874
doi:
Types de publication
Journal Article
Langues
eng
Pagination
4065-4078Informations de copyright
© 2022 Islamuddin et al.
Déclaration de conflit d'intérêts
The authors report no conflict of interest in this work.
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