Paper-based ELISA diagnosis technology for human brucellosis based on a multiepitope fusion protein

• Published in: PLoS neglected tropical diseases 2021-08, Vol.15 (8), p.e0009695-e0009695
• Main Authors: Yin, Dehui, Bai, Qiongqiong, Wu, Xiling, Li, Han, Shao, Jihong, Sun, Mingjun, Jiang, Hai, Zhang, Jingpeng
• Format: Article
• Language: English
• Subjects: Agglutination tests; Animal population; Animal populations; Antibodies; Antigenic determinants; Antigens; Antigens, Bacterial - analysis; Antigens, Bacterial - genetics; Antigens, Bacterial - immunology; Bacteria; Bacterial Outer Membrane Proteins - analysis; Bacterial Outer Membrane Proteins - genetics; Bacterial Outer Membrane Proteins - immunology; Biology and Life Sciences; Biotechnology industry; Brucella; Brucella - genetics; Brucella - immunology; Brucella - isolation & purification; Brucellosis; Brucellosis - diagnosis; Brucellosis - microbiology; China; Developing countries; Diagnosis; Diagnostic systems; Effluents; ELISA; Engineering and Technology; Enzyme-linked immunosorbent assay; Enzyme-Linked Immunosorbent Assay - instrumentation; Enzyme-Linked Immunosorbent Assay - methods; Enzymes; Epitopes; Epitopes - analysis; Epitopes - genetics; Epitopes - immunology; Fusion protein; Health risks; Humans; Identification and classification; Infectious diseases; Laboratories; LDCs; Lipopolysaccharides; Medical research; Medicine and Health Sciences; Membrane proteins; Methods; Molecular diagnostic techniques; Outer membrane proteins; Packaging; Peptides; Populations; Proteins; Research and Analysis Methods; Sensitivity; Sensitivity and Specificity; Specificity; Testing; Tropical diseases; Zinc oxide; Zoonoses
• ISSN: 1935-2735; 1935-2727; 1935-2735
• DOI: 10.1371/journal.pntd.0009695

Brucellosis, as a serious zoonotic infectious disease, has been recognized as a re-emerging disease in the developing countries worldwide. In china, the incidence of brucellosis is increasing each year, seriously threatening the health of humans as well as animal populations. Despite a quite number of diagnostic methods currently being used for brucellosis, innovative technologies are still needed for its rapid and accurate diagnosis, especially in area where traditional diagnostic is unavailable. In this study, a total of 22 B cell linear epitopes were predicted from five Brucella outer membrane proteins (OMPs) using an immunoinformatic approach. These epitopes were then chemically synthesized, and with the method of indirect ELISA (iELISA), each of them displayed a certain degree of capability in identifying human brucellosis positive sera. Subsequently, a fusion protein consisting of the 22 predicted epitopes was prokaryotically expressed and used as diagnostic antigen in a newly established brucellosis testing method, nano-ZnO modified paper-based ELISA (nano-p-ELISA). According to the verifying test using a collection of sera collected from brucellosis and non-brucellosis patients, the sensitivity and specificity of multiepitope based nano-p-ELISA were 92.38% and 98.35% respectively. The positive predictive value was 98.26% and the negative predictive value was 91.67%. The multiepitope based fusion protein also displayed significantly higher specificity than Brucella lipopolysaccharide (LPS) antigen. B cell epitopes are important candidates for serologically testing brucellosis. Multiepitope fusion protein based nano-p-ELISA displayed significantly sensitivity and specificity compared to Brucella LPS antigen. The strategy applied in this study will be helpful to develop rapid and accurate diagnostic method for brucellosis in human as well as animal populations.