Abstract
A new electrochemical method for the quantitation of bacteria that is rapid, inexpensive, and amenable to miniaturization is reported. Cyclic voltammetry was used to quantitate M. luteus, C. sporogenes, and E. coli JM105 in exponential and stationary phases, following exposure of screen-printed carbon working electrodes (SPCEs) to lysed culture samples. Ferricyanide was used as a probe. The detection limits (3s) were calculated and the dynamic ranges for E. coli (exponential and stationary phases), M. luteus (exponential and stationary phases), and C. sporogenes (exponential phase) lysed by lysozyme were 3 × 104 to 5 × 106 colony-forming units (CFU) mL−1, 5 × 106 to 2 × 108 CFU mL−1 and 3 × 103 to 3 × 105 CFU mL−1, respectively. Good overlap was obtained between the calibration curves when the electrochemical signal was plotted against the dry bacterial weight, or between the protein concentration in the bacterial lysate. In contrast, unlysed bacteria did not change the electrochemical signal of ferricyanide. The results indicate that the reduction of the electrochemical signal in the presence of the lysate is mainly due to the fouling of the electrode by proteins. Similar results were obtained with carbon-paste electrodes although detection limits were better with SPCEs. The method described herein was applied to quantitation of bacteria in a cooling tower water sample.
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References
Postgate JR (1969) Methods Microbiol 1:611–628
Rompré A, Servais P, Baudart J, de-Roubin, M-R, Laurent P (2002) J Microbiol Methods 49:31–54
Hafner F (2000) Biosens Bioelectron 15:149–158
Lorenzelli L, Margesin B, Martinoia S, Tedesco MT, Valle M (2003) Biosens Bioelectron 18:621–626
Richards JCS, Jason AC, Hobbs G, Gibson DM, Christie RH (1978) J Phys 11:560–568
Iijima S, Yamashita S, Matsunaga K, Miura H, Morikawa M (1987) J Chem Technol Biotechnol 40:203–213
Ortmanis A, Patterson WI, Neufeld RJ (1991) Enzyme Microb Technol 13:450–455
Karl D (1980) Microbiol Rev 44:739–796
Venkateswaran K, Hattori N, La Duc MT, Kern R (2003) J Microbiol Methods 52:367–377
Selan L, Berlutti F, Passariello C, Thaller MC, Renzini G (1992) J Clin Microbiol 30:1739–1742
La Duc MT, Kern R, Venkateswaran K (2004) Microb Ecol 47:150–158
Seydel JK, Wempe E (1980) Drug Res 30:298–301
Delanghe JR, Kouri TT, Huber AR, Hannemann-Pohl K, Guder WG, Lun A, Sinha P, Stamminger G, Beier L (2000) Clin Chim Acta 301:1–18
Lyons SR, Griffen AL, Leys EJ (2000) J Clin Microbiol 38:2362–2365
Bach H-J, Tomanova J, Schloter M, Munch JC (2002) J Microbiol Methods 49:235–245
Créach V, Baudoux, A-C, Bertru G, Le Rouzic B (2003) J Microbiol Methods 52:19–28
Turner APF, Ramsey G, Higgins IJH (1983) Biochem Soc Trans 11:445–448
Pérez FG, Mascini M, Tothill IE, Turner APF (1998) Anal Chem 70:2380–2386
Morisaki H, Sugimoto M, Shiraishi H (2000) Bioelectrochemistry 51:21–25
Zhao X, Hillard LR, Mechery SJ, Wang Y, Bagwe RP, Jin S, Tan W (2004) PNAS 101:15027–15032
Wang H (2002) J AOAC Int 85:996–999
Han MJ, Lee SY (2006) Microbiol Mol Biol Rev 70:362–439
VanBogelen RA, Schiller EE, Thomas JD, Neidhardt FC (1999) Electrophoresis 20:2149–2159
Ingraham JL, Maaloe O, Neidhardt FC (1983) Growth of the bacterial cell. Sinauer, Sunderland, Massachusetts, pp 2–5
Hanegraaf PPF, Muller EB (2001) J Theor Biol 212:237–251
Umbarger HE (1977) Biochem Educ 5:67–71 (and references therein)
Smiechowski MF, Lvovich VF, Roy S, Fleischman A, Fissell WH, Riga AT (2006) Biosens Bioelectron 22:670–677
Bernabeu P, de Cesare A, Caprani A (1989) J Electroanal Chem 265:261–275
Bernabeu P, Caprani A (1990) Biomaterials 11:258–264
Wright JEI, Cosman NP, Fatih K, Omanovic S, Roscoe SG (2004) J Electroanal Chem 564:185–197
Leger C, Elliott SJ, Hoke KR, Jeuken LJC, Jones AK, Armstrong FA (2003) Biochemistry 42:8653–8662
Moulton SE, Barisci JN, Bath A, Stella R, Wallace GG (2003) J Colloid Interface Sci 261:312–319
Arkoub IA, Randriamahazaka H, Nigretto J-M (1997) Anal Chim Acta 340:99–108
Bianco P (2002) Rev Mol Biotechnol 82:393–409
Fernandez-Sanchez C, Gonzalez-Garcia MB, Costa-Garcia A (2000) Biosens Bioelectron 14:917–924
Lucarelli F, Marrazza G, Turner APF, Mascini M (2004) Biosens Bioelectron 19:515–530
Galan-Vidal C, Munoz J, Domingez C, Algeret S (1995) Trends Anal Chem 14:225–231
Lewis BD (1992) Clin Chem 38:2093–2095
Wang J (1994) Analyst 119:763–766
Wang J, Pedrero M, Cai X (1995) Analyst 120:1969–1972
Skladal P, Morozova NO, Reshetilov AN (2002) Biosens Bioelectron 17:867–873
Rohm I, Genrich M, Collier W, Bilitewski U (1996) Analyst 121:877–881
Bonnet C, Andreescu S, Marty J-L (2003) Anal Chim Acta 481:209–211
Wang J, Tian B, Nascimento VB, Angnes L (1998) Electrochim Acta 43:3459–3465
Mathie AJ (1998) Chemical treatment for cooling water. Fairmont Press, Lilburn, GA, p 19
Winn WC (1988) Clin Microbiol Rev 1:60–81
Millar BC, Jiru X, Moore JE, Earle JAP (2000) J Microbiol Methods 42:139–147
Witholt B, Heerikhizen HV, Leij LD (1976) Biochim Biophys Acta 443:534–544
Compton SJ, Jones CG (1985) Anal Biochem 151:369–374
Christophersen J (1968) In: Hawthorn J, Rolfe EJ (eds) Low temperature biology of foodstuffs. Pergamon Press, New York, pp 251–269
Morrin A, Killard AJ, Smyth MR (2003) Anal Lett 36:2021–2039
Wiatr CL Analyst, Spring 2002: Detection and Eradication of a Non-Legionella Pathogen in a Cooling Water System http://www.awt.org/members/publications/analyst/2002/spring/water_system.htm
Sarkar D, Chattoraj DK (1994) Colloids Surf 2:411–417
Suttiprasit P, McGuire J (1992) J Colloid Interface Sci 154:327–336
Ingraham JL, Maaloe O, Neidhardt FC (1983) Growth of the bacterial cell. Sinauer, Sunderland, Massachusetts, chap 1, pp 1–48
Hajra S, Chattoraj DK (1991) Indian J Biochem Biophys 28:267–279
Gani SA, Mukherjee DC, Chattoraj DK (1999) Langmuir 15:7130–7138
McCoy J (1983) The chemical treatment of cooling water. Chemical Publishing, New York, NY, p 112
Engler CR (1990) Cell disruption by homogenizer. In: Asengo JD (ed) Separation processes in biotechnology. Marcel Dekker, New York
Fykse EM, Olsen JS, Skogan G (2003) J Microbiol Methods 55:1–10
Lee S-W, Tai Y-C (1999) Sens Actuators 73:74–79
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Funding from the Natural Sciences and Engineering Research Council of Canada and the University of Waterloo is gratefully acknowledged.
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Obuchowska, A. Quantitation of bacteria through adsorption of intracellular biomolecules on carbon paste and screen-printed carbon electrodes and voltammetry of redox-active probes. Anal Bioanal Chem 390, 1361–1371 (2008). https://doi.org/10.1007/s00216-007-1825-7
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DOI: https://doi.org/10.1007/s00216-007-1825-7