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<p>The decaprenylphosphoryl-β-D-ribose-2′-epimerase (DprE1) plays a key role in the synthesis of arabinogalactan and lipoarabinomannan makes it a promising target for the development of novel anti-tuberculosis drugs. The flavoenzyme DprE1, encoded by the gene DprE1, is overexpressed in multi-drug-resistant tuberculosis (MDR-TB). This study aims to develop an electrocatalytically active, label-free gene-DprE1 Based biosensor for the determination of MDR-TB using Graphite-Zinc Oxide (Graphite-ZnO) composite nanofibers synthesized with electrospinning technique. To identify its structural morphology, the various characterizations like scanning electron microscopy (SEM), X-ray diffraction (XRD), X-ray photoelectron spectroscopy (XPS), and transmission electron microscopy (TEM) were performed. These nanofibers were further immobilized on the glassy carbon electrodes (GCE) with MSA-EDC-NHS protocol and gene-DprE1 probe and target. The electrocatalytic signal transduction arising from the synergistic interaction between conductive graphite and semiconducting ZnO was systematically evaluated using cyclic voltammetry (CV), differential pulse voltammetry (DPV), and electrochemical impedance spectroscopy (EIS). To evaluate the real-world applicability of the developed biosensor, gene-DprE1 was spiked into human urine samples in order to mimic clinically relevant conditions. This strategy was adopted to determine whether the biosensor could effectively detect the target gene in a non-invasive biological matrix, a feature that holds significant promise for improving diagnostic approaches. Monitoring gene-DprE1 in urine is especially useful since it offers a patient-friendly alternative to traditional intrusive collection procedures. This makes it easier to find MDR-TB early. A wide linear detection range was obtained, extending from 1 pM/mL to 1 nM/mL, making it appropriate for monitoring trace-level biomarkers. The biosensor also showed great sensitivity, with very low limits of detection (LOD) of 0.239 pM/mL using CV, 0.366 pM/mL using DPV, and 0.194 pM/mL using EIS. These results clearly show that the biosensor might be a very reliable, sensitive, and non-invasive diagnostic tool, which makes it even more useful for detecting MDR-TB early in clinical settings.</p> Graphical Abstract <p></p>

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Graphite–ZnO Nanofiber-Decorated GCE for Label-Free Gene-DprE1 Sensing: Toward Rapid MDR-TB Diagnosis

  • 期刊论文

  • 发表日期：

    2026年3月6日

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  • Dinesh R. Rotake,
  • Jitendra B. Zalke,
  • Arpita Parakh,
  • Shubham C Anjankar,
  • Shiv Govind Singh,
  • Ranjana Singh

摘要

The decaprenylphosphoryl-β-D-ribose-2′-epimerase (DprE1) plays a key role in the synthesis of arabinogalactan and lipoarabinomannan makes it a promising target for the development of novel anti-tuberculosis drugs. The flavoenzyme DprE1, encoded by the gene DprE1, is overexpressed in multi-drug-resistant tuberculosis (MDR-TB). This study aims to develop an electrocatalytically active, label-free gene-DprE1 Based biosensor for the determination of MDR-TB using Graphite-Zinc Oxide (Graphite-ZnO) composite nanofibers synthesized with electrospinning technique. To identify its structural morphology, the various characterizations like scanning electron microscopy (SEM), X-ray diffraction (XRD), X-ray photoelectron spectroscopy (XPS), and transmission electron microscopy (TEM) were performed. These nanofibers were further immobilized on the glassy carbon electrodes (GCE) with MSA-EDC-NHS protocol and gene-DprE1 probe and target. The electrocatalytic signal transduction arising from the synergistic interaction between conductive graphite and semiconducting ZnO was systematically evaluated using cyclic voltammetry (CV), differential pulse voltammetry (DPV), and electrochemical impedance spectroscopy (EIS). To evaluate the real-world applicability of the developed biosensor, gene-DprE1 was spiked into human urine samples in order to mimic clinically relevant conditions. This strategy was adopted to determine whether the biosensor could effectively detect the target gene in a non-invasive biological matrix, a feature that holds significant promise for improving diagnostic approaches. Monitoring gene-DprE1 in urine is especially useful since it offers a patient-friendly alternative to traditional intrusive collection procedures. This makes it easier to find MDR-TB early. A wide linear detection range was obtained, extending from 1 pM/mL to 1 nM/mL, making it appropriate for monitoring trace-level biomarkers. The biosensor also showed great sensitivity, with very low limits of detection (LOD) of 0.239 pM/mL using CV, 0.366 pM/mL using DPV, and 0.194 pM/mL using EIS. These results clearly show that the biosensor might be a very reliable, sensitive, and non-invasive diagnostic tool, which makes it even more useful for detecting MDR-TB early in clinical settings.

Graphical Abstract