Nanoscale detection of bacteriophage triggered ion cascade (Invited Paper)

In an era of potential bioterrorism and pandemics of antibiotic-resistant microbes, bacterial contaminations of food and water supplies is a major concern. There is an urgent need for the rapid, inexpensive and specific identification of bacteria under field conditions. Here we describe a method that combines the specificity and avidity of bacteriophages with fluctuation analysis of electrical noise. The method is based on the massive, transitory ion leakage that occurs at the moment of phage DNA injection into the host cell. The ion fluxes require only that the cells be physiologically viable (i.e., have energized membranes) and can occur within seconds after mixing the cells with sufficient concentrations of phage particles. To detect these fluxes, we have constructed a nano-well, a lateral, micron-size capacitor of titanium electrodes with gap size of 150 nm, and used it to measure the electrical field fluctuations in microliter (mm3) samples containing phage and bacteria. In mixtures where the analyte bacteria were sensitive to the phage, large stochastic waves with various time and amplitude scales were observed, with power spectra of approximately 1/f2 shape over at 1 - 10 Hz. Development of this SEPTIC (SEnsing of Phage-Triggered Ion Cascades) technology could provide rapid detection and identification of live, pathogenic bacteria on the scale of minutes, with unparalleled specificity. The method has a potential ultimate sensitivity of 1 bacterium/microliter (1 bacterium/mm3).

[1]  M. Schwartz,et al.  The adsorption of coliphage lambda to its host: effect of variations in the surface density of receptor and in phage-receptor affinity. , 1976, Journal of molecular biology.

[2]  V. Braun,et al.  A common receptor protein for phage T5 and colicin M in the outer membrane of Escherichia coli B. , 1973, Biochimica et biophysica acta.

[3]  Phage DNA transport across membranes. , 1999 .

[4]  P. Boulanger,et al.  Involvement of ion channels in the transport of phage DNA through the cytoplasmic membrane of E. coli. , 1989, Biochimie.

[5]  L. Randall-Hazelbauer,et al.  Isolation of the Bacteriophage Lambda Receptor from Escherichia coli , 1973, Journal of bacteriology.

[6]  Laszlo B. Kish,et al.  Surface diffusion enhanced chemical sensing by surface acoustic waves , 2003 .

[7]  Laszlo B. Kish,et al.  Extracting information from noise spectra of chemical sensors: single sensor electronic noses and tongues , 2000 .

[8]  P. Boulanger,et al.  Characterization of ion channels involved in the penetration of phage T4 DNA into Escherichia coli cells. , 1988, The Journal of biological chemistry.

[9]  J. Smulko,et al.  Fluctuation-enhanced sensing , 2007, SPIE International Symposium on Fluctuations and Noise.

[10]  R. Jacob Baker,et al.  CMOS Circuit Design, Layout, and Simulation , 1997 .

[11]  R. Young,et al.  Oligohistidine Tag Mutagenesis of the λ Holin Gene , 1998, Journal of bacteriology.

[12]  C. Dekker,et al.  Direct measurement of electrical transport through DNA molecules , 2000, Nature.

[13]  A. Lindberg.  Bacteriophage receptors. , 1973, Annual review of microbiology.

[14]  R. Hendrix,et al.  Evolutionary relationships among diverse bacteriophages and prophages: all the world's a phage. , 1999, Proceedings of the National Academy of Sciences of the United States of America.

[15]  D. Rhoden,et al.  API system: a multitube micromethod for identification of Enterobacteriaceae. , 1972, Applied microbiology.

[16]  R. Bender,et al.  Direct Selection for P1-Sensitive Mutants of Enteric Bacteria , 1974, Journal of bacteriology.

[17]  V. C. Bode,et al.  Events in lambda injection between phage adsorption and DNA entry. , 1976, Virology.

[18]  A membrane potential threshold for phage T4 DNA injection. , 1980, Biochemical and biophysical research communications.

[19]  J. A. Connelly,et al.  Low noise electronic system design , 1993 .

[20]  B. Bachmann,et al.  Pedigrees of some mutant strains of Escherichia coli K-12. , 1972, Bacteriological reviews.

[21]  K. Mullis,et al.  Specific synthesis of DNA in vitro via a polymerase-catalyzed chain reaction. , 1987, Methods in enzymology.

[22]  P. Boulanger,et al.  Ion channels are likely to be involved in the two steps of phage T5 DNA penetration into Escherichia coli cells. , 1992, The Journal of biological chemistry.

[23]  D. Kaiser,et al.  Gene transfer to myxobacterium by Escherichia coli phage P1. , 1975, Science.

[24]  S. Silver,et al.  Cation Fluxes and Permeability Changes Accompanying Bacteriophage Infection of Escherichia coli , 1968, Journal of virology.

[25]  R. Hendrix,et al.  Bacteriophage lambda PaPa: not the mother of all lambda phages. , 1992, Science.

[26]  L. Grinius,et al.  Studies on energy supply for genetic processes. Requirement for membrane potential in Escherichia coli infection by phage T4. , 2005, European journal of biochemistry.