WO2005081002A2 - Method for biological identification using high temperature superconductor enhanced nuclear quadrupole resonance - Google Patents

Method for biological identification using high temperature superconductor enhanced nuclear quadrupole resonance Download PDF

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Publication number
WO2005081002A2
WO2005081002A2 PCT/US2004/035319 US2004035319W WO2005081002A2 WO 2005081002 A2 WO2005081002 A2 WO 2005081002A2 US 2004035319 W US2004035319 W US 2004035319W WO 2005081002 A2 WO2005081002 A2 WO 2005081002A2
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biological agent
quadrupole
coils
nanoparticle
nuclei
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PCT/US2004/035319
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French (fr)
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WO2005081002A3 (en
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Daniel B. Laubacher
James D. Mccambridge
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E.I Dupont De Nemours And Company
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    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01R—MEASURING ELECTRIC VARIABLES; MEASURING MAGNETIC VARIABLES
    • G01R33/00—Arrangements or instruments for measuring magnetic variables
    • G01R33/20—Arrangements or instruments for measuring magnetic variables involving magnetic resonance
    • G01R33/44—Arrangements or instruments for measuring magnetic variables involving magnetic resonance using nuclear magnetic resonance [NMR]
    • G01R33/441—Nuclear Quadrupole Resonance [NQR] Spectroscopy and Imaging
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01V—GEOPHYSICS; GRAVITATIONAL MEASUREMENTS; DETECTING MASSES OR OBJECTS; TAGS
    • G01V3/00—Electric or magnetic prospecting or detecting; Measuring magnetic field characteristics of the earth, e.g. declination, deviation
    • G01V3/14—Electric or magnetic prospecting or detecting; Measuring magnetic field characteristics of the earth, e.g. declination, deviation operating with electron or nuclear magnetic resonance

Definitions

  • This invention relates to the use of high temperature superconductor enhanced nuclear quadrupole resonance to detect biological agents.
  • NQR nuclear quadrupole resonance
  • NQR nuclear magnetic resonance
  • RF radio frequency
  • the average power of the applied oscillating magnetic field in a NQR detection system is about 100 Watts, and it can thus operate from standard alternating current (AC) electrical outlets (either 110V or 240V) .
  • AC alternating current
  • HTS high temperature superconductor
  • HTS self-resonant receive coils i.e. sensors
  • S/N signal-to-noise ratio
  • HTS enhanced NQR systems have been scientifically demonstrated for the detection of macroscopic samples of contraband such as explosives and drugs.
  • the object of this invention is to use HTS enhanced NQR systems to provide a method for detecting biological agents, particularly those that could be used for bioterrorism.
  • this invention provides a method for detecting one or more targeted biological agents in air by: (a) collecting a biological agent from the air and concentrating the biological agent in a liquid to form a sample thereof; (b) placing functionalized nanoparticles in the liquid with the biologicalagent , wherein the functionalized nanoparticles contain nuclei with a quadrupole moment and wherein the functionalized nanoparticles attach to the one or more targeted biological agents to produce assembled nanoparticle-biological agent composites,- (c) filtering and drying the assembled nanoparticle-biological agent composites to form a nanoparticle-biological agent sample; (d) placing the nanoparticle-biological agent sample in a NQR spectrometer, the NQR spectrometer comprising one or more high temperature superconductor coils that serve both as excitation coils to provide a radio frequency magnetic field to excite the quadrupole nuclei in the nanoparticles of the nanoparticle-biological agent sample and receive coils to detect
  • this invention provides a method for detecting one or more targeted biological agents in air by: (a) collecting a biological agent from the air and concentrating the biological agent in a liquid to form a sample thereof; (b) placing functionalized nanoparticles in the liquid with the biological sample, wherein the functionalized nanoparticles contain nuclei with a quadrupole moment and wherein the functionalized nanoparticles attach to the one or more targeted biological agents to produce assembled nanoparticle-biological agent composites; (c) filtering and drying the assembled nanoparticle-biological agent composites to form a nanoparticle-biological agent sample; (d) placing the nanoparticle-biological agent sample in a NQR spectrometer, the NQR spectrometer comprising: (i) one or more coils used solely as excitation coils to provide a radio frequency magnetic field to excite the quadrupole nuclei in the nanoparticles of the nanoparticle-biological agent sample, and (ii) one or more coils used solely as receive
  • this invention provides a method for detecting one or more targeted biological agents in air by: (a) collecting a biological agent to form a sample thereof; (b) forming assembled nanoparticle-biological agent composites using functionalized nanoparticles and said biological agent contained in the sample, wherein the nanoparticles contain quadrupole nuclei; (c) placing the nanoparticle-biological agent composites in a NQR spectrometer; and (d) using the NQR spectrometer to excite the quadrupole nuclei and detect the quadrupole NQR signal and thereby confirm the presence of the one or more targeted biological agents.
  • this invention involves a method for detecting one or more targeted biological agents using a NQR spectrometer by: (a) preparing nanoparticle-biological agent composites, wherein the nanoparticles contain quadrupole nuclei ,- (b) placing the nanoparticle-biological agent composites in the NQR spectrometer; and (c) using the NQR spectrometer to excite the quadrupole nuclei and detect the quadrupole NQR signal and thereby confirm the presence of the one or more targeted biological agents .
  • the method of this invention addresses the biggest challenge for a biological agent detection system, the ability to pick out a specific signal from the targeted biological agent while minimizing any signals originating from nonpathogenic biological background.
  • a critical key to the performance of a system based upon HTS enhanced NQR is the ability to miniaturize various critical RF components, such as the HTS self-resonant receive coils, sufficiently to detect small amounts of a biological agent collected during a short collection and analysis time such as a 2 minute collection and analysis time.
  • the biological agents of interest are typically microorganisms that can cause physical harm to or disease among personnel, animals or plants . They can also cause the deterioration of material. These agents generally fall into two broad categories - pathogens (usually called germs) and toxins. Pathogens are living microorganisms that cause lethal or incapacitating diseases. Bacteria, rickettsiae, fungi and viruses are included in the pathogens. Toxins are poisons that plants, animals or microorganisms produce naturally. Possible biological war-fare toxins include a variety of neurotoxic (affecting the central nervous system) and cytotoxic (causing cell death) compounds.
  • a sample of a biological agent can be obtained in a variety of ways suc as by using one of the existing commercial aerosol collection and concentration technologies, e . g.
  • BioCaptiveTM MesoSystems, Kennewick, WA 99336. These systems typically pass large volumes of air through the collection system and disperse the biological agent in a small volume of liquid such as water to form the sample.
  • a sample is prepared in a cyclone collector the larger particles, i.e. the particles of a biological agent, are collected on the outer wall as a result of centrifugal force and then fall into a container. The smaller particles remain in the air stream leaving through the air outlet.
  • These current systems are approximately 50% efficient in capture and concentration of most bioterrorism agents with airflow rates up to 1000 liters per minute.
  • this step may be carried out by simply placing the biological agent in a small amount of liquid such as water or any liquid that does not destroy the biological agent or interfere with the attachment of the biological agent to a functional nanoparticle. Typically, about 10 mL or less of liquid can be used.
  • Functionalized nanoparticles which have at least one dimension on the order of nanometers, are then added to the liquid sample containing the biological agent. Functionalized nanoparticles are prepared so that they attach themselves to the targeted biological agent, e.gr. a virus, toxin, or bacterial spore.
  • This attachment is based, for example, on nucleic acid, antibody/antigen binding or ligand/receptor interactions, and provides assembled nanoparticle- biological agent composites.
  • the nanoparticles are "functionalized” by treating their surfaces with a biological or chemical entity known to bind to the targeted biological agent.
  • gold nanoparticles are used to make functionalized nanoparticles.
  • Nanoparticles can, however, be made from materials containing other metals such as aluminum, cobalt, copper or antimony.
  • Functionalized nanoparticles and the biological sample can also be mixed in the dry state or in a gel . Mixing in a liquid is preferred, however, to provide a more rapid and complete attachment of the functionalized nanoparticles to the biological agent .
  • the functionalized nanoparticles can be of only one type that attaches to only one targeted biological agent, or can be a variety of types that attach to a variety of targeted biological agents.
  • the use of functionalized nanoparticles is a critical key to reducing the effect of nonpathogenic biological background in the environment .
  • the assembled nanoparticle-biological agent composites are filtered and, if they were prepared in liquid, dried to form a nanoparticle-biological agent sample.
  • Any separation process can be used that separates the assembled nanoparticle-biological agent composites from the nanoparticles that do not form a composite with the biological agent.
  • a separation system based on size is suitable because the assembled nanoparticle-biological agent composites are typically larger than the nanoparticles that do not form a composite with the biological agent .
  • the nanoparticles that do not form a composite with the biological agent can be re-used or disposed of after they have been separated from the sample .
  • the sample of the nanoparticle-biological agent, containing the assembled nanoparticle-biological agent composites, is then placed in a NQR spectrometer.
  • the presence of nanoparticle-biological agent composites in the nanoparticle-biological agent is not visibly discernable.
  • the NQR spectrometer is comprised of at least one excitation coil and at least one high temperature superconductor receive coil .
  • a NQR spectrometer can have one or more coils that serve as both excitation and receive coils or it can have separate coils that only excite and only receive.
  • An excitation coil, i.e. transmit coil, of an NQR detection system provides a radio frequency (RF) magnetic field that excites the quadrupole nuclei in a sample, and results in their producing their characteristic resonance signals that the receive coil detects.
  • RF radio frequency
  • the quadrupole nuclei are in the nanoparticles of the nanoparticle-biological agent sample.
  • the quadrupole nuclei can be chosen from any of the nuclei with spin greater than or equal to 1, i.e. from nuclei that have a quadrupole moment. It is preferred to use an element for which the quadrupole nucleus is a predominant isotope of that element.
  • a functionalized nanoparticle that attaches to that targeted biological agent is needed.
  • detection of the nuclear quadrupole resonance frequency of the quadrupole nuclei in the nanoparticles of the nanoparticle-biological agent sample confirms the presence of the targeted biological agent.
  • a functionalized nanoparticle that attaches to each of the different targeted biological agents is needed. If each functionalized nanoparticle that attaches to at least one of the different targeted biological agents, contains the same quadrupole nuclei, detection of an NQR signal will indicate that one or more of the targeted biological agents is present .
  • each functionalized nanoparticle attaches to a different targeted biological agent, and each nanoparticle contain a different quadrupole nuclei .
  • each nanoparticle contain a different quadrupole nuclei .
  • nanoparticle-biological agent composites present in the sample, i.e. no quadrupole nuclei to excite, and therefore no NQR signal will be detected.
  • the nanoparticles will be removed from the sample by the separation process just as will those nanoparticles that do not for some reason form a composite with the biological agent when it is present in the sample.
  • the nanoparticles that do form a composite will remain in the sample after the separation process, and it is the detection the quadrupole nuclei in those nanoparticles that will indicate the presence of the biological agent .
  • a biological sample can be obtained by collecting some of the agent with a small scoop. Again the functionalized nanoparticles and the biological sample can be mixed in the dry state, in a gel or in a liquid so that the functionalized nanoparticles can attach to the biological agent to form assembled nanoparticle- biological agent composites. A liquid is preferred. Once the nanoparticle-biological agent composites are formed, they are placed in a NQR spectrometer to detect the one or more biological agents as described previously.
  • a receive coil i.e. a sensor, made of a high temperature superconductor (HTS) rather than copper since the HTS self-resonant coil has a quality factor Q of the order of 10 3 -10 6 .
  • the NQR signals have low intensity and short duration.
  • S/N signal-to-noise ratio
  • the signal-to-noise ratio is proportional to the square root of Q so that the use of a HTS self-resonant coil as a receive coil results In an increase in S/N by a factor of 10-100 over that of a copper coil.
  • the use of a high temperature superconductor receive coil with a large Q provides a distinct advantage over the use of an ordinary conductor coil .
  • the measurements of the method of this invention are enabled by the high Q and extremely low losses of the HTS receive coils.
  • the excitation coils can be made of copper, silver, aluminum or a high temperature superconductor.
  • a copper, silver or aluminum coil is preferably in the form of a shielded-loop resonator (SLR) coil. SLR's have been developed to eliminate the detuning effect of the electrical interaction between the coil and the surrounding material.
  • SLR shielded-loop resonator
  • the receive' coils are high temperature superconductor coils.
  • the coil is a high temperature superconductor coil .
  • a high temperature superconductor coil is preferably in the form of a self-resonant planar coil, i.e. a surface coil, with a coil configuration of HTS on one or both sides of a substrate .
  • High temperature superconductors are those that superconduct above 77K.
  • the high temperature superconductors used to form the HTS self-resonant coil are preferably selected from the group consisting of YBa 2 Cu 3 0 7 , Tl 2 Ba 2 CaCu 2 0 8 , TlBa 2 Ca 2 Cu 3 O g , (TlPb) Sr 2 CaCu 2 0 7 and (TlPb) Sr 2 Ca 2 Cu 3 O g .
  • the high temperature superconductor is YBa 2 Cu 3 0 7 or Tl 2 Ba 2 CaCu 2 0 8

Abstract

This invention relates to a method for identifying biological agents using high temperature superconductor enhanced nuclear quadrupole resonance. The targeted biological agents are attached to functionalized nanoparticles and the quadrupole nuclei in these nanoparticles are detected by the nuclear quadrupole resonance detection system.

Description

TITLE METHOD FOR BIOLOGICAL IDENTIFICATION USING HIGH TEMPERATURE SUPERCONDUCTOR ENHANCED NUCLEAR QUADRUPOLE RESONANCE
Field of the Invention This invention relates to the use of high temperature superconductor enhanced nuclear quadrupole resonance to detect biological agents.
Background of the Invention With the increased concern regarding bioterrorism and the need to promote homeland security, it is critical to have a method for the rapid detection and • identification of biological agents that could be used for terrorism.
The use of nuclear quadrupole resonance (NQR) as a l means of detecting explosives and other contraband has been recognized for some time, see e . g. T. Hirshfield et al , J. Molec. Struct . 58, 63 (1980), A.N. Garroway et al , Proc . SPIE 2092, 318 (1993), and A.N. Garroway et al , IEEE Trans , on Geoscience and Remote Sensing 39, 1108 (2001) . NQR provides some distinct advantages over other detection methods. NQR spectroscopy can detect and discriminate specific solid state materials containing quadrupole nuclei, i.e. nuclei with a quadrupole moment. As with nuclear magnetic resonance (NMR) , a radio frequency coil produces an oscillating magnetic field at a frequency identified with a specific compound containing one or more quadrupole nuclei. If the specific compound is present a return NQR signal is detected. Unlike NMR, NQR requires no . static magnetic field. NQR spectroscopy is extremely cost effective since it is a radio frequency (RF) technique in a frequency region where silicon based electronics operate. NQR detection systems use non- ionizing magnetic fields near 5 MHz, and therefore present no health hazards to nearby workers and will not damage materials sensitive to ionizing radiation. The average power of the applied oscillating magnetic field in a NQR detection system is about 100 Watts, and it can thus operate from standard alternating current (AC) electrical outlets (either 110V or 240V) . A number of fundamental problems exist for the conventional approach to NQR spectroscopy. These problems result in low throughput and long measurement times. The use of high temperature superconductor (HTS) self-resonant structures, particularly HTS self- resonant receive coils, i.e. sensors, dramatically improves the sensitivity of the NQR spectrometer. For example, the high quality factor Q of an HTS sensor results in a 5-20 dB improvement in the signal-to-noise ratio (S/N) as compared to a conventional sensor. This is important in view of the low intensity NQR signal. HTS enhanced NQR systems have been scientifically demonstrated for the detection of macroscopic samples of contraband such as explosives and drugs. The object of this invention is to use HTS enhanced NQR systems to provide a method for detecting biological agents, particularly those that could be used for bioterrorism.
Summary of the Invention In one embodiment, this invention provides a method for detecting one or more targeted biological agents in air by: (a) collecting a biological agent from the air and concentrating the biological agent in a liquid to form a sample thereof; (b) placing functionalized nanoparticles in the liquid with the biologicalagent , wherein the functionalized nanoparticles contain nuclei with a quadrupole moment and wherein the functionalized nanoparticles attach to the one or more targeted biological agents to produce assembled nanoparticle-biological agent composites,- (c) filtering and drying the assembled nanoparticle-biological agent composites to form a nanoparticle-biological agent sample; (d) placing the nanoparticle-biological agent sample in a NQR spectrometer, the NQR spectrometer comprising one or more high temperature superconductor coils that serve both as excitation coils to provide a radio frequency magnetic field to excite the quadrupole nuclei in the nanoparticles of the nanoparticle-biological agent sample and receive coils to detect the nuclear quadrupole resonance signal from the quadrupole nuclei; and (e) using the NQR spectrometer to excite the quadrupole nuclei and detect the quadrupole NQR signal and thereby confirm the presence of the one or more targeted biological agents .
In another embodiment, this invention provides a method for detecting one or more targeted biological agents in air by: (a) collecting a biological agent from the air and concentrating the biological agent in a liquid to form a sample thereof; (b) placing functionalized nanoparticles in the liquid with the biological sample, wherein the functionalized nanoparticles contain nuclei with a quadrupole moment and wherein the functionalized nanoparticles attach to the one or more targeted biological agents to produce assembled nanoparticle-biological agent composites; (c) filtering and drying the assembled nanoparticle-biological agent composites to form a nanoparticle-biological agent sample; (d) placing the nanoparticle-biological agent sample in a NQR spectrometer, the NQR spectrometer comprising: (i) one or more coils used solely as excitation coils to provide a radio frequency magnetic field to excite the quadrupole nuclei in the nanoparticles of the nanoparticle-biological agent sample, and (ii) one or more coils used solely as receive coils to detect the nuclear quadrupole resonance signal from the quadrupole nuclei; and (e) using the NQR spectrometer to excite the quadrupole nuclei and detect the quadrupole NQR signal and thereby confirm the presence of the one or more targeted biological agents .
In still another embodiment, this invention provides a method for detecting one or more targeted biological agents in air by: (a) collecting a biological agent to form a sample thereof; (b) forming assembled nanoparticle-biological agent composites using functionalized nanoparticles and said biological agent contained in the sample, wherein the nanoparticles contain quadrupole nuclei; (c) placing the nanoparticle-biological agent composites in a NQR spectrometer; and (d) using the NQR spectrometer to excite the quadrupole nuclei and detect the quadrupole NQR signal and thereby confirm the presence of the one or more targeted biological agents.
In yet another embodiment, this invention involves a method for detecting one or more targeted biological agents using a NQR spectrometer by: (a) preparing nanoparticle-biological agent composites, wherein the nanoparticles contain quadrupole nuclei ,- (b) placing the nanoparticle-biological agent composites in the NQR spectrometer; and (c) using the NQR spectrometer to excite the quadrupole nuclei and detect the quadrupole NQR signal and thereby confirm the presence of the one or more targeted biological agents .
Description of the Preferred Embodiments The method of this invention addresses the biggest challenge for a biological agent detection system, the ability to pick out a specific signal from the targeted biological agent while minimizing any signals originating from nonpathogenic biological background.
The technology that makes this possible is high temperature superconductivity enhanced nuclear quadrupole resonance. This technology has not previously been utilized for detecting biological agents .
A critical key to the performance of a system based upon HTS enhanced NQR is the ability to miniaturize various critical RF components, such as the HTS self-resonant receive coils, sufficiently to detect small amounts of a biological agent collected during a short collection and analysis time such as a 2 minute collection and analysis time. The biological agents of interest are typically microorganisms that can cause physical harm to or disease among personnel, animals or plants . They can also cause the deterioration of material. These agents generally fall into two broad categories - pathogens (usually called germs) and toxins. Pathogens are living microorganisms that cause lethal or incapacitating diseases. Bacteria, rickettsiae, fungi and viruses are included in the pathogens. Toxins are poisons that plants, animals or microorganisms produce naturally. Possible biological war-fare toxins include a variety of neurotoxic (affecting the central nervous system) and cytotoxic (causing cell death) compounds.
When the biological agent is airborne, a sample of a biological agent can be obtained in a variety of ways suc as by using one of the existing commercial aerosol collection and concentration technologies, e . g.
BioCaptive™, MesoSystems, Kennewick, WA 99336. These systems typically pass large volumes of air through the collection system and disperse the biological agent in a small volume of liquid such as water to form the sample. When a sample is prepared in a cyclone collector the larger particles, i.e. the particles of a biological agent, are collected on the outer wall as a result of centrifugal force and then fall into a container. The smaller particles remain in the air stream leaving through the air outlet. These current systems are approximately 50% efficient in capture and concentration of most bioterrorism agents with airflow rates up to 1000 liters per minute. If the commercial equipment does not form a sample by dispersing the biological agent in a liquid, this step may be carried out by simply placing the biological agent in a small amount of liquid such as water or any liquid that does not destroy the biological agent or interfere with the attachment of the biological agent to a functional nanoparticle. Typically, about 10 mL or less of liquid can be used. Functionalized nanoparticles, which have at least one dimension on the order of nanometers, are then added to the liquid sample containing the biological agent. Functionalized nanoparticles are prepared so that they attach themselves to the targeted biological agent, e.gr. a virus, toxin, or bacterial spore. This attachment is based, for example, on nucleic acid, antibody/antigen binding or ligand/receptor interactions, and provides assembled nanoparticle- biological agent composites. The nanoparticles are "functionalized" by treating their surfaces with a biological or chemical entity known to bind to the targeted biological agent. Typically, gold nanoparticles are used to make functionalized nanoparticles. Nanoparticles can, however, be made from materials containing other metals such as aluminum, cobalt, copper or antimony. Functionalized nanoparticles and the biological sample can also be mixed in the dry state or in a gel . Mixing in a liquid is preferred, however, to provide a more rapid and complete attachment of the functionalized nanoparticles to the biological agent .
The functionalized nanoparticles can be of only one type that attaches to only one targeted biological agent, or can be a variety of types that attach to a variety of targeted biological agents. The use of functionalized nanoparticles is a critical key to reducing the effect of nonpathogenic biological background in the environment .
The assembled nanoparticle-biological agent composites are filtered and, if they were prepared in liquid, dried to form a nanoparticle-biological agent sample. Any separation process can be used that separates the assembled nanoparticle-biological agent composites from the nanoparticles that do not form a composite with the biological agent. A separation system based on size is suitable because the assembled nanoparticle-biological agent composites are typically larger than the nanoparticles that do not form a composite with the biological agent . The nanoparticles that do not form a composite with the biological agent can be re-used or disposed of after they have been separated from the sample .
The sample of the nanoparticle-biological agent, containing the assembled nanoparticle-biological agent composites, is then placed in a NQR spectrometer. In view of the small quantity of biological material collected, the presence of nanoparticle-biological agent composites in the nanoparticle-biological agent is not visibly discernable.
The NQR spectrometer is comprised of at least one excitation coil and at least one high temperature superconductor receive coil . A NQR spectrometer can have one or more coils that serve as both excitation and receive coils or it can have separate coils that only excite and only receive. An excitation coil, i.e. transmit coil, of an NQR detection system provides a radio frequency (RF) magnetic field that excites the quadrupole nuclei in a sample, and results in their producing their characteristic resonance signals that the receive coil detects.
In this invention, the quadrupole nuclei are in the nanoparticles of the nanoparticle-biological agent sample. The quadrupole nuclei can be chosen from any of the nuclei with spin greater than or equal to 1, i.e. from nuclei that have a quadrupole moment. It is preferred to use an element for which the quadrupole nucleus is a predominant isotope of that element.
If only one targeted biological agent is to be detected, only one type of functionalized nanoparticle, i.e. a functionalized nanoparticle that attaches to that targeted biological agent, is needed. In this case, detection of the nuclear quadrupole resonance frequency of the quadrupole nuclei in the nanoparticles of the nanoparticle-biological agent sample confirms the presence of the targeted biological agent. If more than one targeted biological agent are to be detected, 'a functionalized nanoparticle that attaches to each of the different targeted biological agents is needed. If each functionalized nanoparticle that attaches to at least one of the different targeted biological agents, contains the same quadrupole nuclei, detection of an NQR signal will indicate that one or more of the targeted biological agents is present . Preferably, however, each functionalized nanoparticle attaches to a different targeted biological agent, and each nanoparticle contain a different quadrupole nuclei . By attaching a nanoparticle containing a specific type of quadrupole nuclei to each specific targeted biological agent, the detection of an NQR signal from that particular quadrupole nuclei will indicate that the specific targeted biological agent attached to the nanoparticle containing that specific type of quadrupole nuclei is present.
In any case, if no targeted biological agent is present, there will be no nanoparticle-biological agent composites present in the sample, i.e. no quadrupole nuclei to excite, and therefore no NQR signal will be detected. If no targeted biological agent is present, the nanoparticles will be removed from the sample by the separation process just as will those nanoparticles that do not for some reason form a composite with the biological agent when it is present in the sample. The nanoparticles that do form a composite will remain in the sample after the separation process, and it is the detection the quadrupole nuclei in those nanoparticles that will indicate the presence of the biological agent .
When the biological agent is available in a more concentrated form, e . g. on a surface or in an envelope, a biological sample can be obtained by collecting some of the agent with a small scoop. Again the functionalized nanoparticles and the biological sample can be mixed in the dry state, in a gel or in a liquid so that the functionalized nanoparticles can attach to the biological agent to form assembled nanoparticle- biological agent composites. A liquid is preferred. Once the nanoparticle-biological agent composites are formed, they are placed in a NQR spectrometer to detect the one or more biological agents as described previously.
It is especially advantageous to use a receive coil, i.e. a sensor, made of a high temperature superconductor (HTS) rather than copper since the HTS self-resonant coil has a quality factor Q of the order of 103-106. The NQR signals have low intensity and short duration. In view of the low intensity NQR signal, it is important to have a signal-to-noise ratio (S/N) as large as possible. The signal-to-noise ratio is proportional to the square root of Q so that the use of a HTS self-resonant coil as a receive coil results In an increase in S/N by a factor of 10-100 over that of a copper coil. Therefore, the use of a high temperature superconductor receive coil with a large Q provides a distinct advantage over the use of an ordinary conductor coil . The measurements of the method of this invention are enabled by the high Q and extremely low losses of the HTS receive coils. When separate coils are used for the excitation and receive coils in the method of this invention, the excitation coils can be made of copper, silver, aluminum or a high temperature superconductor. A copper, silver or aluminum coil is preferably in the form of a shielded-loop resonator (SLR) coil. SLR's have been developed to eliminate the detuning effect of the electrical interaction between the coil and the surrounding material.
When separate coils are used for the excitation and receive coils in the method of' this invention, the receive' coils are high temperature superconductor coils. When a single coil is used as both an excitation and a receive coil in the method of this invention, the coil is a high temperature superconductor coil . A high temperature superconductor coil is preferably in the form of a self-resonant planar coil, i.e. a surface coil, with a coil configuration of HTS on one or both sides of a substrate . High temperature superconductors are those that superconduct above 77K. The high temperature superconductors used to form the HTS self-resonant coil are preferably selected from the group consisting of YBa2Cu307, Tl2Ba2CaCu208, TlBa2Ca2Cu3Og, (TlPb) Sr2CaCu207 and (TlPb) Sr2Ca2Cu3Og. Most preferably, the high temperature superconductor is YBa2Cu307 or Tl2Ba2CaCu20 8

Claims

CLAIMSWhat is claimed is:
1. A method for detecting one or more targeted biological agents in air, comprising: (a) collecting a biological agent from the air and concentrating the biological agent in a liquid to form a sample thereof; (b) placing functionalized nanoparticles in the liquid with the biologicalagent , wherein the functionalized nanoparticles contain nuclei with a quadrupole moment and wherein the functionalized nanoparticles attach to the one or more targeted biological agents to produce assembled nanoparticle-biological agent composites; (c) filtering and drying the assembled nanoparticle-biological agent composites to form a nanoparticle-biological agent sample ; (d) placing the nanoparticle-biological agent sample in a NQR spectrometer, the NQR spectrometer comprising one or more high temperature superconductor coils that serve both as excitation coils to provide a radio frequency magnetic field to excite the quadrupole nuclei in the nanoparticles of the nanoparticle-biological agent sample and receive coils to detect the nuclear quadrupole resonance signal from the quadrupole nuclei; and (e) using the NQR spectrometer to excite the quadrupole nuclei and detect the quadrupole NQR signal and thereby confirm the presence of the one or more targeted biological agents .
2. The method of claim 1, wherein two or more targeted biological agents are to be detected, and the functionalized nanoparticles are selected so that the functionalized nanoparticles that attach to each different targeted biological agent contain different quadrupole nuclei, thereby insuring that there is a specific type of quadrupole nuclei attached to each different targeted biological agent.
3. The method of claim 1 or 2 , wherein the one or more high temperature superconductor coils are high temperature superconductor self-resonant planar coils.
4. The method of claim 3 , wherein the NQR spectrometer further comprises one or more high temperature superconductor self-resonant planar coils used solely as receive coils. i
5. A method for detecting one or more targeted biological agents in air, comprising: (a) collecting a biological agent from the air and concentrating the biological agent in a liquid to form a sample thereof; (b) placing functionalized nanoparticles in the liquid with the biologicalagent , wherein the functionalized nanoparticles contain nuclei with a quadrupole moment and wherein the functionalized nanoparticles attach to the one or more targeted biological agents to produce assembled nanoparticle-biological agent composites; (c) filtering and drying the assembled nanoparticle-biological agent composites to form a nanoparticle-biological agent sample; (d) placing the nanoparticle-biological agent sample in a NQR spectrometer, the NQR spectrometer comprising: (i) one or more coils used solely as excitation coils to provide a radio frequency magnetic field to excite the quadrupole nuclei in the nanoparticles of the nanoparticle-biological agent sample, and (ii) one or more high temperature superconductor coils used solely as receive coils to detect the nuclear quadrupole resonance signal from the quadrupole nuclei; and (e) using the NQR spectrometer to excite the quadrupole nuclei and detect the quadrupole NQR signal and thereby confirm the presence of the one or more targeted biological agents .
6. The method of claim 5 , wherein the one or more coils used solely as excitation coils are shielded-loop resonator coils.
7. The method of claim 5, wherein the one or more coils used solely as excitation coils are high temperature superconductor coils.
8. The method of claim 5, wherein two or more targeted biological agents are to be detected, and the functionalized nanoparticles are selected so that the functionalized nanoparticles that attach to each different targeted biological agent contain different quadrupole nuclei, thereby insuring that there is a specific type of quadrupole nuclei attached to each different targeted biological agent.
9. The method of any of claims 5-7, wherein the high temperature superconductor coils are high temperature superconductor self-resonant planar coils.
10. The method of claim 9, wherein two or more targeted biological agents are to be detected, and the functionalized nanoparticles are selected so that the functionalized nanoparticles that attach to each different targeted biological agent contain different quadrupole nuclei, thereby insuring that there is a specific type of quadrupole nuclei attached to each different targeted biological agent.
11. A method for detecting one or more targeted biological agents using a NQR spectrometer, comprising: (a) preparing nanoparticle-biological agent composites, wherein the nanoparticles contain quadrupole nuclei; (b) placing the nanoparticle-biological agent composites in the NQR spectrometer; and (c) using the NQR spectrometer to excite the quadrupole nuclei and detect the quadrupole NQR signal and thereby confirm the presence of the one or more targeted biological agents .
12. A method for detecting one or more targeted biological agents, comprising: (a) collecting a biological agent to form a sample thereof ,- (b) forming assembled nanoparticle-biological agent composites using functionalized nanoparticles and the biological agent contained in the sample, wherein the nanoparticles contain quadrupole nuclei; (c) placing the nanoparticle-biological agent composites in a NQR spectrometer; and (d) using the NQR spectrometer to excite the quadrupole nuclei and detect the quadrupole NQR signal and thereby confirm the presence of the one or more targeted biological agents.
13. The method of claim 12 , wherein two or more targeted biological agents are to be detected, and the functionalized nanoparticles used to form the assembled nanoparticle-biological agent composites are selected so that functionalized nanoparticles that attach to each different biological agent contain different quadrupole nuclei, thereby insuring that there is a specific type of quadrupole nuclei attached to each different targeted biological agent.
14. The method of claim 12, wherein the NQR spectrometer is comprised of one or more high temperature superconductor coils that serve both as excitation coils to provide a radio frequency magnetic field to excite the quadrupole nuclei in the nanoparticles of the nanoparticle-biological agent composites and receive coils to detect the nuclear quadrupole resonance signal from the quadrupole nuclei .
15. The method of claim 12, wherein the NQR spectrometer is comprised of: (i) one or more coils used solely as excitation coils to provide a radio frequency magnetic field to excite the quadrupole nuclei in the nanoparticles of the nanoparticle-biological agent composites, and (ii) one or more high temperature superconductor coils used solely as receive coils to detect the nuclear quadrupole resonance signal from the quadrupole nuclei.
16. The method of Claim 12 wherein the nanoparticle comprises gold.
PCT/US2004/035319 2003-10-23 2004-10-22 Method for biological identification using high temperature superconductor enhanced nuclear quadrupole resonance WO2005081002A2 (en)

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Families Citing this family (9)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US7521932B2 (en) * 2003-05-06 2009-04-21 The Penn State Research Foundation Method and system for adjusting the fundamental symmetric mode of coupled high temperature superconductor coils
US7511496B2 (en) * 2006-02-27 2009-03-31 The Penn State Research Foundation Quadrupole resonance using narrowband probes and continuous wave excitation
WO2007100760A2 (en) 2006-02-27 2007-09-07 The Penn State Research Foundation Detecting quadrupole resonance signals using high temperature superconducting resonators
US20070211922A1 (en) * 2006-03-10 2007-09-13 Crowley Christopher W Integrated verification and screening system
US7868758B2 (en) * 2006-03-10 2011-01-11 Morpho Detection, Inc. Passenger screening system and method
US20080018451A1 (en) * 2006-07-11 2008-01-24 Jason Benfielt Slibeck Passenger screening system and method
US8022703B1 (en) * 2010-05-06 2011-09-20 Kai-Wen Huang Method for rapid detecting tumor
CN103474171B (en) * 2012-06-07 2015-08-26 清华大学 The preparation method of superconducting wire
WO2016106098A1 (en) * 2014-12-23 2016-06-30 One Resonance Sensors, Llc Apparatus and method for detecting explosives

Citations (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO1992019978A1 (en) * 1987-05-11 1992-11-12 Datalight Limited Portable nmr and nqr spectrometers
WO1999045409A1 (en) * 1998-03-06 1999-09-10 Btg International Ltd. Nqr testing method and apparatus
WO2000070356A1 (en) * 1999-05-19 2000-11-23 Intermagnetics General Corporation Magnetically equivalent rf coil arrays
WO2002098364A2 (en) * 2001-06-06 2002-12-12 The General Hospital Corporation Magnetic-nanoparticle conjugates and methods of use
WO2003014700A2 (en) * 2001-08-08 2003-02-20 Theragenics Corporation Use of isotopes to provide information to an article
US6566873B1 (en) * 1996-08-28 2003-05-20 Btg International Limited Method of and apparatus for nuclear quadrupole resonance testing a sample

Family Cites Families (58)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US4514691A (en) 1983-04-15 1985-04-30 Southwest Research Institute Baggage inspection apparatus and method for determining presences of explosives
GB8711114D0 (en) 1987-05-11 1987-06-17 Jonsen P Spectrometers
US5168224A (en) 1988-10-07 1992-12-01 Hitachi, Ltd. Detecting method and apparatus of specific substance
GB2289344B (en) 1991-04-02 1996-01-17 British Tech Group Detection method and apparatus
GB9106789D0 (en) 1991-04-02 1991-05-22 Nat Res Dev Nqr methods and apparatus
US5206592A (en) 1991-05-23 1993-04-27 Buess Michael L Detection of explosives by nuclear quadrupole resonance
US5233300A (en) 1991-05-23 1993-08-03 The United States Of America As Represented By The Secretary Of The Navy Detection of explosive and narcotics by low power large sample volume nuclear quadrupole resonance (NQR)
GB9112290D0 (en) 1991-06-07 1991-07-24 Nat Res Dev Methods and apparatus for nqr imaging
GB2286248B (en) 1991-06-07 1995-11-22 British Tech Group Detection method and apparatus
US6335622B1 (en) 1992-08-25 2002-01-01 Superconductor Technologies, Inc. Superconducting control elements for RF antennas
US5262394A (en) 1991-12-27 1993-11-16 The United States Of America As Represented By The United States Department Of Energy Superconductive articles including cerium oxide layer
US5585723A (en) 1995-03-23 1996-12-17 Conductus, Inc. Inductively coupled superconducting coil assembly
US5565778A (en) 1992-06-01 1996-10-15 Conductus, Inc. Nuclear magnetic resonance probe coil
US5276398A (en) 1992-06-01 1994-01-04 Conductus, Inc. Superconducting magnetic resonance probe coil
US5351007A (en) 1992-06-01 1994-09-27 Conductus, Inc. Superconducting magnetic resonance probe coil
GB9319875D0 (en) 1993-09-27 1994-03-09 British Tech Group Apparatus for and methods of nuclear resonance testing
WO1995034096A1 (en) 1994-06-03 1995-12-14 E.I. Du Pont De Nemours And Company Fluoropolymer protectant layer for high temperature superconductor film and photo-definition thereof
JP3828573B2 (en) 1994-09-29 2006-10-04 ビーティージー・インターナショナル・リミテッド Nuclear quadrupole resonance test method and method of configuring nuclear quadrupole resonance test apparatus
US5592083A (en) 1995-03-08 1997-01-07 Quantum Magnetics, Inc. System and method for contraband detection using nuclear quadrupole resonance including a sheet coil and RF shielding via waveguide below cutoff
US5594338A (en) 1995-03-08 1997-01-14 Quantum Magnetics, Inc. Automatic tuning apparatus and method for substance detection using nuclear quadrupole resonance and nuclear magnetic resonance
DE19513231A1 (en) 1995-04-07 1996-10-10 Siemens Ag Antenna. e.g. for medical NMR
GB9508635D0 (en) 1995-04-28 1995-06-14 Mansfield Peter Method and apparatus for elimination of mutual coupling in magnetic coils
US5750473A (en) 1995-05-11 1998-05-12 E. I. Du Pont De Nemours And Company Planar high temperature superconductor filters with backside coupling
US5656937A (en) 1995-06-07 1997-08-12 Conductus, Inc. Low-noise symmetric dc SQUID system having two pairs of washer coils and a pair of Josephson junctions connected in series
CA2226263C (en) 1995-07-11 2007-08-14 British Technology Group Limited Apparatus for and method of nuclear quadrupole testing of a sample
US5571146A (en) 1995-10-31 1996-11-05 Pacesetter, Inc. Technique for welding dissimilar metals
US5777474A (en) 1996-11-08 1998-07-07 Advanced Imaging Research, Inc. Radio-frequency coil and method for resonance imaging/analysis
US5804967A (en) 1996-11-15 1998-09-08 The United States Of America As Represented By The Secretary Of The Navy Apparatus and method for generating short pulses for NMR and NQR processing
US6242918B1 (en) 1996-11-15 2001-06-05 The United States Of America As Represented By The Secretary Of The Navy Apparatus and method for reducing the recovery period of a probe in pulsed nuclear quadrupole resonance and nuclear magnetic resonance detection systems by varying the impedance of a load to reduce total Q factor
AU5369398A (en) 1996-12-02 1998-06-29 Trustees Of Columbia University In The City Of New York, The Multiple resonance superconducting probe
WO1998037438A1 (en) 1997-02-25 1998-08-27 Advanced Imaging Research, Inc. Radio-frequency coil array for resonance analysis
CA2290626A1 (en) 1997-05-27 1998-12-03 Murali K. Cherukuri Resonant structure for spatial and spectral-spatial imaging of free radical spin probes using radiofrequency time domain electron paramagnetic resonance spectroscopy
US6201392B1 (en) 1997-11-07 2001-03-13 Varian, Inc. Coplanar RF probe coil arrangement for multifrequency excitation
US6025719A (en) 1997-11-07 2000-02-15 Varian, Inc. Nuclear magnetic resonance method and apparatus
US6420872B1 (en) 1998-01-13 2002-07-16 The United States Of America As Represented By The Secretary Of The Navy Probe for detecting a transient magnetic resonance signal, wherein the ratio of the Q of the probe to the Q of the resonance signal is relatively large
AU3267499A (en) 1998-03-06 1999-09-20 Btg International Limited Apparatus for and method of nuclear quadrupole resonance testing a sample in thepresence of interference
US6218943B1 (en) 1998-03-27 2001-04-17 Vivid Technologies, Inc. Contraband detection and article reclaim system
US6054856A (en) 1998-04-01 2000-04-25 The United States Of America As Represented By The Secretary Of The Navy Magnetic resonance detection coil that is immune to environmental noise
US6108569A (en) 1998-05-15 2000-08-22 E. I. Du Pont De Nemours And Company High temperature superconductor mini-filters and mini-multiplexers with self-resonant spiral resonators
US6104190A (en) 1998-11-17 2000-08-15 The United States Of America As Represented By The Secretary Of The Navy Nuclear quadrupole resonance (NQR) method and apparatus for detecting a nitramine explosive
US6336822B1 (en) * 1999-01-26 2002-01-08 Veam S.R.L. Handle operated power connector
GB9915842D0 (en) 1999-07-06 1999-09-08 Btg Int Ltd Methods and apparatus for analysing a signal
US6291994B1 (en) 2000-01-14 2001-09-18 Quantum Magnetics, Inc. Active Q-damping sub-system using nuclear quadrupole resonance and nuclear magnetic resonance for improved contraband detection
JP3551899B2 (en) 2000-06-26 2004-08-11 株式会社村田製作所 Resonator, filter, duplexer and communication device
US6556013B2 (en) 2001-03-09 2003-04-29 Bruker Biospin Corp. Planar NMR coils with localized field-generating and capacitive elements
DE10118835C2 (en) 2001-04-17 2003-03-13 Bruker Biospin Ag Faellanden Superconducting resonators for applications in NMR
US7091721B2 (en) 2001-04-18 2006-08-15 IGC—Medical Advances, Inc. Phased array local coil for MRI imaging having non-overlapping regions of sensitivity
US6900633B2 (en) 2001-07-02 2005-05-31 The United States Of America As Represented By The Secretary Of The Navy Substance detection by nuclear quardrupole resonance using at least two different excitation frequencies
US6590394B2 (en) 2001-09-28 2003-07-08 Varian, Inc. NMR probe with enhanced power handling ability
AUPR868201A0 (en) 2001-11-05 2001-11-29 Thorlock International Limited Q-factor switching method and apparatus for detecting nuclear quadrupole and nuclear magnetic resonance signals
USD459245S1 (en) 2001-11-26 2002-06-25 Garrett Electronics, Inc. Hand-held metal detector
US6617591B1 (en) * 2001-12-03 2003-09-09 Sandia Corporation Method for remote detection of trace contaminants
US7247865B2 (en) * 2001-12-31 2007-07-24 Lockheed Martin Corporation System and method of detecting, neutralizing, and containing suspected contaminated articles
AUPS224702A0 (en) 2002-05-10 2002-06-13 Thorlock International Limited Transmit - receive coil system for nuclear quadrupole resonance signal detection in substances
US6819109B2 (en) 2003-01-23 2004-11-16 Schonstedt Instrument Company Magnetic detector extendable wand
US20040222790A1 (en) 2003-02-18 2004-11-11 Ntzo Inc. Method and apparatus for threat screening of step-on and laid-on items
US6777937B1 (en) 2003-03-06 2004-08-17 The United States Of America As Represented By The Secretary Of The Navy Nuclear quadrupole resonance method and apparatus
US6952163B2 (en) * 2003-06-11 2005-10-04 Quantum Magnetics, Inc. Combined systems user interface for centralized monitoring of a screening checkpoint for passengers and baggage

Patent Citations (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO1992019978A1 (en) * 1987-05-11 1992-11-12 Datalight Limited Portable nmr and nqr spectrometers
US6566873B1 (en) * 1996-08-28 2003-05-20 Btg International Limited Method of and apparatus for nuclear quadrupole resonance testing a sample
WO1999045409A1 (en) * 1998-03-06 1999-09-10 Btg International Ltd. Nqr testing method and apparatus
WO2000070356A1 (en) * 1999-05-19 2000-11-23 Intermagnetics General Corporation Magnetically equivalent rf coil arrays
WO2002098364A2 (en) * 2001-06-06 2002-12-12 The General Hospital Corporation Magnetic-nanoparticle conjugates and methods of use
WO2003014700A2 (en) * 2001-08-08 2003-02-20 Theragenics Corporation Use of isotopes to provide information to an article

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