Screening and identification of immunoactive peptide mimotopes for the enhanced serodiagnosis of tuberculosis

Abstract

Although serological detection is a practical strategy for early detection and diagnosis of tuberculosis (TB), inconsistent and imprecise estimates of sensitivity and specificity block its development and application for clinic. New or alternative serological antigens with improved accuracy are urgently needed. A phage-displayed random peptide library was employed to screen for immunoactive peptides using specific immunoglobulin G (IgG) of TB patients as target molecules. With two screening strategies, 20 single phages displaying different sequences were obtained and no sequence homology was found among these phages. From the results of phage-ELISA, H12, TB6, TB15, and TB18 phages showed higher affinity to IgGs from TB patients(S/N ≥2.1) and were identified as the positive clones. Significant differences in the detection values of sera from 47 TB patients and 37 healthy individuals were found for these four phage clones. According to the reactivity of 284 human sera to synthetic H12, TB6, TB15, and TB18 peptides as determined by ELISA, TB15 showed significantly higher areas under the curve (AUC) and sensitivity than other peptides, providing a lead molecule for the development of new serology diagnostic strategies for TB.

Virus-Derived Peptides for Clinical Applications

“…Recently, phage-display technology was used to develop a short peptide, WHLPLSL, capable of serologically detecting Myobacterium tuberculosis to diagnose tuberculosis. 70 The peptide was examined for its detection ability (in an ELISA test) using the sera from 47 tuberculosis patients and 37 healthy individuals. This research group had also determined from previous work that an antigen epitope peptide of a protein called mycobacterium protein 64 and a protein called CFP10/ESAT-6 could serve as an alternative corresponding antigen for the serological detection of tuberculosis.…”

“…Plates were coated with 4 μ g of peptides per well. 70 Adapted with permission from ref 70 . Copyright 2016 Springer.…”

“…These data could serve as a comparison to see if the new detection method with the peptide WHLPLSL was superior to the previously established method (which sometimes failed to detect tuberculosis) of detecting the proteins mycobacterium protein 64 and CFP10/ESAT-6. 70,164 It was demonstrated that the peptide, WHLPLSL, was a sensitive detection probe for the diagnosis of tuberculosis. Additionally, 3 other peptides showed promise but were not as effective as WHLPLSL in an ELISA.…”

“…Additionally, 3 other peptides showed promise but were not as effective as WHLPLSL in an ELISA. 70 Effectiveness was measured in terms of specificity (ability to correctly identify those without tuberculosis, who are the negative controls) and sensitivity (ability to correctly identify those patients with tuberculosis, who are the positive controls). Therefore, the best treatment option would have both a high sensitivity and a high specificity.…”

“…Therefore, alternative strategies are needed. Recently, phage-display technology was used to develop a short peptide, WHLPLSL, capable of serologically detecting Myobacterium tuberculosis to diagnose tuberculosis . The peptide was examined for its detection ability (in an ELISA test) using the sera from 47 tuberculosis patients and 37 healthy individuals.…”