Publication: Experimental and in silico studies on bioengineered polyester nanobeads for latent tuberculosis diagnosis
| dc.contributor.author | Shaffee, Nurul Syahidah | |
| dc.date.accessioned | 2025-11-16T07:11:20Z | |
| dc.date.available | 2025-11-16T07:11:20Z | |
| dc.date.issued | 2025-08 | |
| dc.description.abstract | Mycobacterium tuberculosis (MTB) is the causative agent for tuberculosis (TB) and remains one of the top ten causes of mortality, with 1.4 million deaths in 2021 alone. With a 50% mortality rate if left untreated, active TB infection needs to be diagnosed as soon as possible. Currently, it is estimated that a quarter of the world's population is infected with latent TB infection (LTBI), that acts as an MTB reservoir. Current diagnostic methods for LTBI, such as the Mantoux Tuberculin Skin Test (TST) and the Interferon Gamma Release Assay (IGRA), have limitations: the former exhibits low specificity, while the latter is expensive to be widely use especially in high TB burden areas. Aiming to decrease the cost of TB infection diagnostic methods, a feasibility study on a new platform (i.e., bioengineered biopolyester nanobeads), using the IGRA technique was conducted. This platform was already successfully tested to diagnose M. bovis-infected cows. The same three MTB antigens (i.e., ESAT-6, CFP-10, and TB7.7) used in IGRA, was utilised to bioengineer the MTB-antigen-coated biopolyester nanobeads platform (BMTB). Optimisation of the BMTB concentration was performed using human samples. The optimised concentration of BMTB was determined to be 10 ng. A comparative analysis of IFN- levels between BMTB and QuantiFERON (QFT) kit, as recommended by the manufacturer, was performed. QFT demonstrated superior performance compared to BMTB in the IFN- release assays. In order to evaluate the discriminatory power of BMTB in three sample groups: pulmonary TB (PTB), TST positive (TSTp), and TST negative (TSTn), the concentrations of six selected cytokines (i.e., IFN-γ, IP-10, IL-2, TNF-α, CCL-3,CCL-11) in them were analysed. IL-2 and CCL-11 were the best-performing individual cytokines for distinguishing PTB against TSTp, TSTn, and healthy individual (TSTp and TSTn) group, achieving greater than 80% sensitivity and specificity. The best cytokine combination is IFN-+IL-2 when PTB is compared to TSTn, TSTp, and to healthy individual (i.e., TSTn + TSTp) groups. Then, to further improve the sensitivity of the BMTB platform, additional antigens that have high immunogenic properties were identified using in silico (immunoinformatics) approaches. These properties are based on six T cell epitope (TCE) criteria: matching with MTB H37Rv antigens, coverage across 273 MTB strains, association with highly expressed MTB genes, promiscuous epitopes, population coverage, and overlap with B cell epitopes (BCEs). Each TCE has a cumulative score of all six criteria. MTB antigens were then ranked based on their corrected total TCE scores within them. Top -ranked antigens were compared to literature for their inferred diagnostic potential. Among the shorlisted antigens are the existing IGRA antigens (ESAT-6 and CFP-10), newly identified potential antigens like ESAT-6-like proteins (i.e., EsxJ, EsxK, EsxM, EsxP, EsxW), the Ag85 family (i.e., Ag85A, Ag85B, and Ag85C), and several others (e.g., HspX, PfkB, and Rv1733). In conclusion, the BMTB showed potential as a low-cost LTBI diagnostic platform, although its sensitivity can be further enhanced by incorporating additional highly immunogenic MTB antigens | |
| dc.identifier.uri | https://erepo.usm.my/handle/123456789/23090 | |
| dc.language.iso | en | |
| dc.subject | - | |
| dc.title | Experimental and in silico studies on bioengineered polyester nanobeads for latent tuberculosis diagnosis | |
| dc.type | Resource Types::text::thesis::doctoral thesis | |
| dspace.entity.type | Publication | |
| oairecerif.author.affiliation | Universiti Sains Malaysia |