Method and apparatus for the detection of volatile products...

Chemistry: molecular biology and microbiology – Apparatus – Including measuring or testing

Reexamination Certificate

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C435S288100, C435S288700, C435S808000, C435S031000, C435S034000, C436S020000, C436S149000, C436S171000, C422S090000, C422S091000, C073S023340, C073S023360, C073S023370

Reexamination Certificate

active

06767732

ABSTRACT:

STATEMENT REGARDING FEDERALLY SPONSORED RESEARCH OR DEVELOPMENT
Not Applicable.
REFERENCE TO A “COMPACT DISC APPENDIX”
Not Applicable.
BACKGROUND OF THE INVENTION
(1) Field of the Invention
The present invention relates to a method and apparatus for detection of volatile products from a sample using a transducer which changes voltage as a function of contact of the volatile products with the transducer to produce a gas signature of the volatile products and a spectrophotometer to analyze the volatile products to produce a spectral footprint of the volatile products. The apparatus and method are used to detect spoilage of a biological material, such as a food. The apparatus is also used to detect microorganisms and by comparing the gas signature and spectral footprint to a library of gas signatures and spectral footprints, the apparatus enables identification of the microorganisms and in particular identification of pathogenic microorganisms.
(2) Description of Related Art
Food safety concerns are currently impacting public health, the meat industry, and animal production agriculture. Animal agriculture has been under increasing scrutiny as a source of foodborne pathogens. There is a need to develop a new technology that can be applied to pre-harvest food safety efforts, particularly for identifying and monitoring a potential human pathogen “on the farm”.
Escherichia coli
(
E. coli
) 0157:H7 has been recognized as a significant bacterial pathogen belonging to a group of enterohemorrhagic
E. coli
associated with bloody diarrhea. It is important public health concern because of its association with commonly consumed foods, such as ground beef. Infection with this organism can cause hemorrhagic colitis, hemolytic uremic syndrome, and thrombotic thrombocytopenic purpura. The association of
E. coli
0157:H7 with ground beef has led to the identification of cattle as a reservoir for the organism. Recent pre-harvest food safety efforts have emphasized identifying factors within cattle production systems for the monitoring and control of
E. coli
0157:H7.
Computer controlled gas sensor based instruments, referred to as artificial olfactory or electronic nose technology, are finding increasing application in the food industry. The sensors are designed to detect volatile compounds that result from spoilage, rancidity, or other “off” odors. Promising results have been shown when this technology was applied to differentiating between different species of bacteria and spoilage fungi.
In today's farming industry, potatoes are stored in large bins before they are shipped out to their various destinations. Disease during storage is magnified due to extended storage periods and by requiring higher storage temperatures for immediate processing of the potatoes (Varns and Glynn, 1979). Disease losses of potatoes in storage may be as high as 30% (The Grower, 1980). As potato processing contributes up to two billion dollars a year to the economy, a small percentage of disease losses represent a significant cost to the potato industry. Currently, the managers of the potato bins monitor odor and wetness at the bottom of the bin to determine rot. By the time these indicators are detected, economic losses can be significant. At the moment, nothing can be done to arrest the spread of the damage. Monitoring of volatiles arising from host-pathogen interactions could become an important early warning of potato disease problems. Disease due to
Erwinia carotovora
infection is a major problem in potato storage.
Erwinia carotovora
is a facultative anaerobic organism, in which the bacterium breaks down the structure of the vegetative cells of infected potatoes, causing soft rot. This causes a layer of wet slime to form on the outside of the potato, resulting in anaerobic conditions in the underlying cells (Costa and Loper, 1994). Varns and Glynn (1979) reported that potatoes infected with the bacterium
Erwinia carotovora
showed high levels of acetone, ethanol, and 2-butanone. Additional volatiles included acetaldehyde, methyl acetate, ethyl acetate, propanethiol, hydrogen sulfide, methyl sulfide, methyl disulfide, n-propanol, and isobutanol (Varns and Glynn, 1979). Waterer and Pritchard (1984) reported methanol, acetaldehyde, ethanol, 2-propanol, acetone, 1-propanol, and 1-butanol in the headspace of
E. carotovora
var.
carotovora
infected Russet Burbank potato tubers. These volatiles can be produced from intermediates as well as the end-product (pyruvate) of the Embden-Meyerhof pathway (Metzler, 1977).
The sense of smell has long been used as a diagnostic tool by medical professionals, law enforcement, food handlers, and countless others in everyday life. The human nose contains approximately 50 million cells in the olfactory epithelium that act as primary receptors to odorous molecules (Gardner et al., 1990; Vandendorpe, 1998). This parallel architecture led to the construction of the electronic nose, which mimics the biological system. The electronic nose is a state-of-the-art technology that can be used to provide rapid and continuous monitoring of a wide array of different volatile compounds. The term “electronic nose” is applied to an array of chemical sensors, where each sensor has only partial specificity to a wide range of odorant molecules (Bartlett et al., Food Technol. 51: 44-48 (1997)). By mapping the sensitivity of the sensors to different chemicals, a complex odor can be “fingerprinted” and identified (Lipman, 1998). The primary receptors in the biological system are replaced by an array of transducers, such as metal oxide films, that respond to a broad range of chemical vapors or odors. Electronic nose instrumentation has advanced rapidly during the past ten years, the majority of application being within the food and drink industries (Gardner and Bartlett, 1992; Kress-Rogers, 1997). Research is being done on the applications of the electronic nose in human healthcare, particularly in the identification of infection (Doctor's Guide to Medical & Other News. “Electronic Nose Sniffs out Infection”). The instrument has also been successfully applied to detect vapors (Gardner et al., 1990; Keller et al., 1994) and aviation fuels (Lauf and Hoffheins, 1990).
Application in microbial detection has been reported for
Clostridium perfringes
, Proteus,
Haemophilus influenzae, Bacteriodes fragilis, Oxford staphylococcus, Pseudomonas aeruginosa
(Craven et al., 1994),
Staphylococcus aureus
, and
E. coli
(Gardner et al., Measurement Sci. Technol. 9: 120-127 (1998)).
U.S. Pat. No. 5,807,701 to Payne et al. provides a method for identifying a microorganism that includes abstracting gas or vapor associated with the microorganism from a detection region and flowing the same over an array of sensors of which an electrical property varies according to exposure to gases or vapors and observing the response of the sensors. An apparatus for detecting a microorganism is also disclosed having a detector means for detecting a gas or vapor associated with the microorganism which includes an array of sensors of which an electrical property varies according to exposure to the gases or vapors.
U.S. Pat. No. 6,017,440 to Lewis et al. provides a sensor array for detecting a microorganism comprising first and second sensors electrically connected to an electrical measuring apparatus, wherein the sensors comprise a region of nonconducting organic material and a region of conducting material that is different than the nonconducting organic material and an electrical path through the regions of nonconducting organic material and the conducting material. Further provided is a system for identifying microorganisms using the sensor array, a computer and a pattern recognition algorithm, such as a neural net are also disclosed.
U.S. Pat. No. 6,244,096 to Lewis et al. provides a device for detecting the presence of an analyte, wherein the analyte is a microorganism marker gas. The device comprises a sample chamber having a fluid inlet port for the influx of the microorganism marker gas; a fluid concentrator i

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