A Nanosensor System Based On Disuccinimydyl–CYP2E1 for Amperometric Detection of the Anti-Tuberculosis Drug, Pyrazinamide
Pyrazinamide (PZA) is among the first-line pro-drugs in the tuberculosis (TB) combination chemotherapy used to treat Mycobacterium tuberculosis. Numerous reports have suggested that hepatotoxicity due to pyrazinamide in patients is due to inappropriate dosing. It is, therefore necessary to develop sensitive and reliable techniques for determining the PZA metabolic profile of diagnosed patients promptly and at point-of-care. This study reports the determination of PZA based on nanobiosensor systems developed from disuccinimidyl octanedioate modified Cytochrome P450-2E1 (CYP2E1) electrodeposited on gold substrates derivatised with (poly(8-anilino-1-napthalene sulphonic acid) PANSA/PVP-AgNPs nanocomposites. The rapid and sensitive amperometric PZA detection gave a dynamic linear range of 2µM to 16µM revealing a limit of detection of 0.044µM and a sensitivity of 1.38µA/µM. The Michaelis-Menten parameters; KM, KM app and IMAX were calculated to be 6.0µM, 1.41µM and 1.51x10-6 A, respectively, indicating a nanobiosensor suitable for use in serum.
Digital Object Identifier (DOI): doi.org/10.5281/zenodo.1090478Procedia APA BibTeX Chicago EndNote Harvard JSON MLA RIS XML ISO 690 PDF Downloads 1910
 M. Zimic, P. Fuentes, R.H. Gilman, A.H. Gutiérrez, D. Kirwan and P. Sheen P, ‘‘Pyrazinoic Acid Efflux Rate in Mycobacterium Tuberculosis is a better proxy of Pyrazinamide Resistance,’’ Tuberculosis (Edinb), vol. 92, no.1, pp. 84–91, January 2012.
 T. Sahota and O.D. Pasqua, ‘‘Feasibility of a Fixed-Dose Regimen of Pyrazinamide and its Impact on Systemic Drug Exposure and Liver Safety in Patients with Tuberculosis,’’ Antimicrob. Agents Chemother., vol. 56, no. 11, pp. 5442-5449, November 2011.
 Y. Zhang, A. Scorpio, H. Nikaido and Z. Sun, ‘‘Role of Acid pH and Defficient Efflux of Pyrazinoic Acid in Unique Susceptibility of Mycobacterium tuberculosis to Pyrazinamide,’’ J. Bacteriol., vol. 181, no.7, pp 2044-2049, April 1999.
 M. Salfinge, and L.B. Heifets, ‘‘Determination of Pyrazinamide MICs for mycobacterium tuberculosis at Different pHs by the Radiometric Method,’’ Antimicrob. Agents Chemot., vol. 32, no. 7, pp.1002-1004, July 1988.
 Y. Zhang, and D. Mitchison, ‘‘The Curious characteristics of pyrazinamide: a review.’’ Int. J. Tuberc. Lung Dis., vol. 7, 1, pp 6-21, January 2003.
 Y. Zhang, M. W. Wade, A. Scorpio, H. Zhang, and Z. Sun, ‘‘Mode of Action of Pyrazinamide: disruption of Mycobacterium tuberculosis membrane transport and energetics by pyrazinoic acid,’’ J. Antimicrob. Chemother., vol. 52, 5, pp. 790-795, November 2003.
 P. Lu, A.C. Haagsma, H. Pham, J.J.Maaksant, S. Mol, H. Lill, and D. Bald, ‘‘Pyrazinoic Acid Decreases the Proton Motive Force, Respiratory ATP Synthesis Activity, and Cellular ATP Levels,’’ Antimicrob. Agents Chemother., vol. 55, no. 11, pp. 5354-5357, November 2011.
 T. Gumbo, C.S. Dona, C. Meek, and R. Leff, ‘‘Pharmacokinetics-Pharmacidynamics of Pyrazinamide in a Novel In Vitro Model of Tuberculosis for Sterilizing Effect: a Paradigm for Faster Assessment of New Antituberculosis Drugs,’’ Antimicrob. Agents Chemother., vol. 53, no. 8, pp. 3197-3204, August 2009.
 W.Shi, X. Zhang, X. Jiang, H. Vuan, J.S. Lee, C.E. Barry 3rd, H. Wang, W. Zhang, and Y. Zhang, ‘‘Pyrazinamide Inhibits Trans-Translation in Mycobacterium tuberculosis,’’ Science, vol. 333, no 6049, pp.1630-1632, September 2011.
 S.T. Cole, ‘‘Pyrazinamide- Old TB Drug Finds New Target,’’ Science, vol. 333, no.6049, pp. 1583-1584, September 2011.
 W. Bleibel, S. Kim, and K. D’Silva, ‘‘Drug-Induced Liver Injury: Review Article,’’ Did. Dis. Science., vol. 52, pp. 2463-2471, March 2007.
 K. C. Chang, C. C. Leung, W. W. Yew, T. Y. Lau, and C.M. Tam, ‘‘Hepatotoxicity of Pyrazinamide,’’ Am. J. Respir. Crit. Care Med., vol. 177, no.12, pp. 13911397, June 2013.
 R. F. Ngece, N. West, P. M. Ndangili, and E. I. Iwuoha, ‘‘A silver Nanoparticle/Poly (8-Anilino-1-Naphthalene Sulphonic Acid) Bioelectrochemical Biosensor System for the Analytical Determination of Ethambutol,’’ Int. J. Electrochem. Sc., vol.6, pp. 1820-1834, June 2011.
 H. Eoh, and P. J. Brennan, ‘‘The Mycobacterium tuberculosis MEP (2C-methyl-D-erythritol 4-phosphate) pathway as a new drug target,’’ Tuberculosis, vol. 89, no. 1, pp.1-11, January 2011.
 W. C. Wang, J. Y. Chen, Y. K. Chen, and L. M. Lin, ‘‘Tuberculosis of the head and neck: a review of 20 cases,’’ Oral Surg. Oral Med. Oral Pathol. Oral Radiol. Endod., vol. 107, no. x, pp. 381-382, month 2009.
 Y. Yin Zhou and L. Zhu, ‘‘Electrochemical behavior of bisphenol A at glassy carbon electrode modified with gold nanoparticles, silk fibroin, and PAMAM dendrimers,’’ Microchim. Acta, vol.170, no. 3, pp. 100–105, March 2010.
 Ding H. Hongbo, and S-M. Park, ‘‘Electrochemistry of Conductive Polymers XXVII. Effects of Polystyrene Sulfonate on Electrochemical Behavior of Polyaniline,’’ Int. J. Electrochem. Sc., vol.150, no. 2, pp. E34-E38, January 2002.
 T. Yamamoto, Y. Moriwaki, and K. Higashino, ‘‘Study of the metabolism of pyrazinamide using a high-performance liquid chromatographic analysis of urine samples,’’ Anal. Biochem., vol. 160, no.2, pp. 346-349, February 1987.
 A. Mehmedagic, P. Verite, S. Menager, D. Andre, and O. Lafont, ‘‘Determination of pyrazinamide and its main metabolites in rat urine by high-performance liquid chromatography,’’ J. Chromatogr. B Biomed. Sci. Appl., vol. 695, no, 2, pp. 365-372, August 1997.