Detection of Dengue Virus-Specific IgM and IgG Antibodies through Peptide Sequences of Envelope and NS1 Proteins for Serological Identification

Abstract

Dengue is an acute febrile illness caused by positive-sense single-stranded RNA virus, belonging to the family Flaviviridae and genus Flavivirus. Transmission of virus among the individuals occurred by blood-feeding Aedes mosquitoes. This virus has four serotypes differentiated on the basis of antibody neutralization assay. At present, there is no particular treatment or vaccine candidate available for dengue infection. Approximately 3.9 billion human populations are at risk of dengue virus (DENV) infection. Thus, precise diagnosis of dengue at the early stage is very essential for disease control and effective therapy in order to treat or prevent severe complications. Indeed, the accurate diagnosis of DENV remains a problem because of low detection accuracy along with high testing price. Sensitivity and specificity of available kits vary from test to test, and cross-reactivity with other Flavivirus is a challenging issue for diagnosis. In this study, linear epitopes of envelope (E) and NS1 proteins were identified to diagnose the DENV. Whole protein sequences of E and NS1 of DENV were obtained from UniProtKB database. On the basis of algorithm prediction from DNASTAR, BCEPRED, and IEDB data resources, twelve peptides of E (EP1 to EP12) and eight peptides of NS1 (NS1-1 to NS1-8) were selected, which were common in all serotypes. Sequence homologies of peptides with other Flavivirus were checked by Multiple Sequence Alignment Tool ClustalX2. Peptide sequences were synthesized chemically by solid-phase peptide synthesis technique. Dengue-specific IgM and IgG (secondary response) antibodies in the patient’s antisera were tested with the peptides using ELISA protocol. Peptides EP1, EP2, EP4, EP7, EP10, and EP12 of E protein and NS1-1, NS1-3, NS1-4, NS1-7, and NS1-8 of NS1 protein were considered the best immunoreactive peptides with the sensitivity (73.33-96.66%) and specificity (82.14-100%). Such peptides together can be used to construct the multiple antigen peptides (MAP) or multiplexed microbeads for designing a precise, cost-effective, and easy-to-make peptide-based immunodiagnostic kit for DENV detection.

Linear and Continuous Flavivirus Epitopes From Naturally Infected Humans

“…In addition, another study predicted and selected common peptides among all DENV serotypes showing no similarities with other flaviviruses from the envelope and Ns1 protein regions of DENV2. Subsequently, these peptides were chemically synthesized using a solid-phase synthesis technique and evaluated by ELISA to perform DENV-specific detection of IgM and IgG from the sera of infected patients (Nagar et al, 2020). This study identified seven continuous linear peptides from the envelope region (Pep34, RCIGISNRDFVEG VSGGSWVDIVL; Pep37, NTTTASRCPTQGEP; Pep43, MENK AWLVHRQWFLDLPLPWLPGADT; Pep53, KEIAETQHG TIVIRVQYEGDG; Pep61, WFKKGSSIGQMFETTMRGA; Pep62, RMAILGDTAWDFGSLGGV; and Pep68, MNSRS TSLSVSQVLVGIVTLYLGV) and four linear peptides from the Ns1 region (Pep102, TEQYKFQPESPSKLASAIQKA; Pep111, ALNDTWKIEKASF; Pep112, PETAECPNTNRAW; and Pep115, VLESEMVIPKNFAGPKSQ).…”

“…Based on prediction algorithms from the DNASTAR, BCEPRED, and IEDB databases, one study selected eight potential diagnostic DENV2 peptides from the Ns1 protein that are common to all DENV serotypes (Nagar et al, 2020). These peptides were chemically synthesized, and their DENV antibody detection levels were evaluated.…”

Peptide Biomarkers for the Diagnosis of Dengue Infection

“…DENV serological diagnosis includes a wide spectrum of different formats and Ag designs (13), however low specificity has constantly been reported for these tests due to the presence of impurities in Ags widely used in commercial tests such as whole viral lysates. Recombinant proteins such as NS1 and Envelope proteins have helped in reducing false positivity and were rapidly adapted to different formats such as indirect ELISA, MAC-ELISA, and lateral flow devices for their commercial application (4,(14)(15)(16)(17). Nevertheless, despite the progress made with the operational characteristics and good sensitivity, challenges with false positivity remain especially with the increasing co-circulation of arthropod-borne flaviviruses across the globe (4,(18)(19)(20)(21).…”

Single-Layer-Graphene-Coated and Gold-Film-Based Surface Plasmon Resonance Prism Coupler Sensor for Immunoglobulin G Detection

“…In theory, the detection of specific antibodies (such as immunoglobulin IgG/IgM) and antigens provides a simple, rapid, reliable, and accessible strategy for the efficient and large-scale screening of suspected cases in point-of-care settings [ 1 , 2 , 3 , 4 ]. Many methods have been proposed for virus infectious disease diagnosis based on the detection of IgG/IgM antibodies [ 5 , 6 , 7 , 8 ]. Jiang et al [ 9 ] presented the label-free, real-time detection of rabbit IgG antibody over a concentration range of 0–100 μg/mL with a resolution of 0.0625 μg/mL.…”