Multiplex peptide microarray profiling of antibody reactivity against neglected tropical diseases derived B-cell epitopes for serodiagnosis in Zimbabwe

Abstract

Introduction Peptides (B-cell epitopes) have broad applications in disease diagnosis and surveillance of pathogen exposure. In this framework, we present a pilot study to design and produce a peptide microarray for the integrated surveillance of neglected tropical diseases. The peptide microarray was evaluated against peptides derived from Ascaris lumbricoides, Necator americanus, Schistosoma haematobium, Schistosoma mansoni, Trichuris trichiura, Bacillus anthracis, Mycobacterium leprae, Wuchereria bancrofti, Rabies lyssavirus, Chlamydia trachomatis and Trypanosoma brucei. Methods S. haematobium was diagnosed using the urine filtration technique. S. mansoni, A. lumbricoides, N. americanus and T. trichiura were diagnosed using the Kato Katz and formal ether concentration techniques. Immunogenic peptides were retrieved from the Tackling Infection to Benefit Africa infectious diseases epitope microarray. Further peptides were predicted using ABCpred. IgG and IgM reactivity against the derived peptides were evaluated using peptide microarray multiplex immunoassays. Positive response was defined as fluorescence intensity ≥ 500 fluorescence units. Immunodominant peptides were identified using color-coded heat maps and bar graphs reflecting the obtained fluorescence signal intensities. Receiver Operating Characteristic analysis and Mann-Whitney-U test were performed to determine the diagnostic validity of the peptides. Results Species-specific responses with at least one peptide derived from each NTD pathogen were observed. The reactive peptides included; for S. haematobium, XP_035588858.1-206-220 and XP_035588858.1-206-220 immunodominant for IgG and IgM respectively, for S. mansoni, P20287.1-58-72 immunodominant for both antibodies and for T. trichiura, CDW52482.1-326-340 immunodominant for IgG and CDW57769.1-2017-2031 and CDW57769.1-1518-1532 immunodominant for IgM. According to ROC analysis most of the peptides selected were inaccurate; with AUC < 0.5. Some peptides had AUC values ranging from 0.5 to 0.5875 for both IgM and IgG suggesting no discrimination. Conclusion Multiplex peptide microarrays are a valuable tool for integrated NTDs surveillance and for screening parasites exposure in endemic areas. Species sero-reactivity observed in the study maybe indicative of exposure to the different NTDs parasites. However, although peptides with the least cross reactivity were selected there is need to validate the sero-reactivity with recombinant antigens and immune-blotting techniques such as western blotting.

Identification ofSchistosoma haematobiumandSchistosoma mansonilinear B-cell epitopes with diagnostic potential usingin silicoimmunoinformatic tools and peptide microarray technology

“…The findings may be attributed to the fact that peptide microarray immunoassays were used in the determination of antibody reactivity against the peptides whilst in other two studies (18,19) antibody reactivity was investigated using peptide based serum IgG ELISA. Pertaining the other 10 published peptides XP_035587815.1-269-283, XP_012797374.1-78-92, AAZ29530.1-25-29, XP_012799745.1-16-30, P20287.1-58-72, AAA29903.1-222-237, P09841.3-6-20, AAA29900.1-145-159, P09792.1-29-43, XP_035588858.1-206-220 none of them showed a clear discrimination between the schistosome infected and uninfected groups and these results are in agreement with findings from our previous study (26).…”

“…Eighteen previously published peptides were included on the peptide microarray, 10 from our previous study Vengesai and colleagues, 2022 (26) and 8 peptides from two studies by Carvalho and Colleagues, 2022 and Lopes and Colleagues, 2017 (18,19). Despite Carvalho and Colleagues, 2022 and Lopes and Colleagues, 2017 showing that peptides Sm168240, Smp_136560 (1564–1578), Smp_126160(438–452), Sm140560, Sm041370, Smp_180240(339–353), Smp_150390.1(216–230), Smp_093840(219–233) had AUC values ranging from 0.76 (95 % CI 0.6024-0.9213) to 0.99 (95 % CI 0.987-100) results from this current study show that only one published peptide Smp 126160-438-452 had an acceptable diagnostic performance with an AUC value of 0.7115 (95% CI 0.605-0.82218).…”

“…Immunoassays were performed by PEPperPRINT GmbH (Heidelberg, Germany) as previously described (26,27). Briefly, the immunoassays involved two steps on the same microarray.…”

Serological diagnosis of soil-transmitted helminth (Ascaris, Trichuris and hookworm) infections: A scoping review

“…Direct and indirect fluorescent antibody tests (FAT) [ 31 , 32 ], radioimmunoassays (RIA) [ 33 , 34 ], complement fixation tests [ 35 , 36 ], and double immunodiffusion tests [ 30 , 34 , 36 , 37 ], were employed until 2003, after which ELISA and western blot became the predominant techniques together with new technologies that appeared on the scene ( S4 Info ). Among these new technologies, the multiplex bead assay [ 38 ], peptide microarray immunoassays [ 39 , 40 ] and quantitative suspension array technology (qSAT) assays [ 41 , 42 ] represent examples of multiplex assays. These advanced technologies have been applied in studies focused on serodiagnosis for multiple STH species ( S3 Info ).…”

Peptide microarray IgM and IgG screening of pre-SARS-CoV-2 human serum samples from Zimbabwe for reactivity with peptides from all seven human coronaviruses: a cross-sectional study

“…These 339 serum samples were collected between Nov 8 and Nov 14, 2019, as part of a larger study investigating the use of serological diagnostic approaches for infectious disease surveillance in Zimbabwe (TIBA Making a Difference project on Multiplex Diagnostics). 13 The 339 samples analysed in our study were from people aged 1–88 years from two rural villages in northeastern Zimbabwe: Mupfure (in the Shamva district; n=214 serum samples) and Murewa (in the Murewa district; n=125 serum samples).…”