Anal. Chem. 2007, 79, 5540-5546
Molecular Commonality Detection Using an
Artificial Enzyme Membrane for in Situ One-Stop
Biosurveillance
Shinya Ikeno,†,‡ Hitoshi Asakawa, and Tetsuya Haruyama*
†
,†,‡
Department of Biological Functions and Engineering, Kyushu Institute of Technology, Kitakyushu Science and
Research Park, Fukuoka, 808-0196, Japan, and CREST, Japan Science and Technology Agency, Kawaguchi,
Saitama, 332-0012, Japan
viruses, is important for maintaining safety and security.1-3 Many
biosurveillance techniques have been developed thus far. Sensor
technology will further contribute to the implementation of
effective biosurveillance. Hygienic applications are expanding
explosively, for example, in food production and hazard analysis
Biodetection and biosensing have been developed based
on the concept of sensitivity toward specific molecules.
However, current demand may require more levelheaded
or far-sighted methods, especially in the field of biological
safety and security. In the fields of hygiene, public safety,
and security including fighting bioterrorism, the detection
of biological contaminants, e.g., microorganisms, spores,
and viruses, is a constant challenge. However, there is
as yet no sophisticated method of detecting such contami-
nants in situ without oversight. The authors focused their
attention on diphosphoric acid anhydride, which is a
structure common to all biological phosphoric substances.
Interestingly, biological phosphoric substances are pecu-
liar substances present in all living things and include
many different substances, e.g., ATP, ADP, dNTP, pyro-
phosphate, and so forth, all of which have a diphosphoric
acid anhydride structure. The authors took this common
structure as the basis of their development of an artificial
enzyme membrane with selectivity for the structure com-
mon to all biological phosphoric substances and studied
the possibility of its application to in situ biosurveillance
sensors. The artificial enzyme membrane-based ampero-
metric biosensor developed by the authors can detect
various biological phosphoric substances, because it has
a comprehensive molecular selectivity for the structure
of these biological phosphoric substances. This in situ
detection method of the common diphosphoric acid
anhydride structure brings a unique advantage to the
fabrication of in situ biosurveillance sensors for monitor-
ing biological contaminants, e.g., microorganism, spores,
and viruses, without an oversight, even if they were
transformed.
4,5
critical control point (HACCP), clinical activities, clinical medi-
cine,6 good manufacturing practice (GMP),8,9 and biological
hazard security, including fighting bioterrorism. In conventional
biosurveillance, the plate culture method has been employed. This
method is appropriate for the detection of living microorganisms,
but it is time-consuming. In other cases of biosurveillance
methods, specific molecular markers are traced by mean of time-
consuming ex situ methods. Specific proteins located on the
surface of microorganisms, spores and viruses are a good markers
that show up readily in affinity-based assays allowing the recogni-
,7
1,10
tion of the species of contaminant.
However, affinity-based
assays can detect only the single contaminant species for which
they are specifically designed. They cannot detect all contaminant
comprehensively. At the frontline of biosurveillance, an in situ
monitoring method of detecting biological contaminants without
oversight is very much required. Conventional methods target
specific marker molecules. This type of strategy can be used to
determine specific species of biological contaminants, but it cannot
be used to perform comprehensive in situ surveys for a strict and
continuous surveillance. In order to perform comprehensive
biosurveillance and keep a strict watch on all biological contami-
nants without any oversight, the modification of the base concept
is probably required.
(
1) Koopmans, M.; Duizer, E. Int. J. Food Microbiol. 2004, 90, 23-41.
(2) Rueckert, A.; Ronimus, R. S.; Morgan, H. W. Food Microbiol. 2006, 23,
20-230.
2
(
3) Booth, T. F.; Kournikakis, B.; Bastien, N.; Ho, J.; Kobasa, D.; Stadnyk, L.;
Li, Y.; Spence, M.; Paton, S.; Henry, B.; Mederski, B.; White, D.; Low, D.
E.; McGeer, A.; Simor, A.; Vearncombe, M.; Downey, J.; Jamieson, F. B.;
Tang, P.; Plummer, F. J. Infect. Dis. 2005, 191, 1472-1477.
4) da Cruz, A. G.; Cenci, S. A.; Maia, M. C. A. Trends Food Sci. Technol. 2006,
17, 406-411.
The social demand for biological safety and security is
currently heightened in many different fields. In the fields of
hygiene, public safety including fighting bioterrorism, the detec-
tion of biological contaminant, e.g., microorganisms, spores, and
(
(
(
(
5) Stecchini, M. L.; Del Torre, M. Vet. Res. Commun. 2005, 29, 117-121.
6) Raspor, P. Acta Biochim. Pol. 2005, 52, 659-664.
7) Cassens, U.; Ahlke, C.; Garritsen, H.; Krakowitzky, P.; Wullenweber,
J.; Fischer, R. J.; Peters, G.; Sibrowski, W. Transfusion 2002, 42,
10-17.
*
To whom correspondence should be addressed, E-mail: haruyama@
life.kyutech.ac.jp, Phone and Fax: +81-(0)93-695-6065.
(8) Plumb, K. Chem. Eng. Res. Des. 2005, 83, 730-738.
(9) Velagaleti, R.; Burns, P. K.; Gill, M. Drug Inf. J. 2003, 37, 407-438.
(10) Chen, F. C.; Godwin, S. L. J. Food Prot. 2006, 69, 2534-2538.
†
Kyushu Institute of Technology.
‡
CREST.
5540 Analytical Chemistry, Vol. 79, No. 15, August 1, 2007
10.1021/ac0708902 CCC: $37.00 © 2007 American Chemical Society
Published on Web 07/07/2007