JP4669259B2 - Test substance analyzer and quantitative method - Google Patents

Description

  The present invention relates to a test substance analyzer and a quantification method. More specifically, the present invention relates to a test substance that quantitatively measures the concentration of a test substance in a sample using magnetic particles as labels and a magnetic sensor as magnetic detection means. The present invention relates to a test substance analyzer and a quantitative method. In particular, it is suitable for medical diagnosis and environmental measurement.

Optical measuring methods such as enzyme immunoassay and fluorescent immunoassay are widely used as methods for measuring the amount of a test substance in a sample.
In the enzyme immunoassay, a substance that specifically reacts with a test substance is labeled with an enzyme, and when the test substance exists, for example, the color development due to the enzyme reaction is optically measured. However, there are problems such as enzyme stability problems and low sensitivity of the detection system.

  In fluorescence immunoassay, a substance that reacts specifically with a test substance is labeled with a fluorescent substance, and the fluorescence intensity from the fluorescent substance in the presence of the test substance is measured to measure the amount of the test substance. Is the method. This fluorescent immunoassay method is generally known as a high-sensitivity measurement method, but is susceptible to stray light and has a problem of generation of background noise and a problem of fading of a phosphor.

  In recent years, a magnetic measurement method for measuring a test substance using a magnetic sensor and magnetic particles has attracted attention with respect to the optical measurement method described above. In the magnetic measurement method, the background noise is generated only from a magnetic substance, and is an effective measurement method with low background noise from the outside as compared with an optical measurement method typified by a fluorescence immunoassay method.

  A superconducting quantum buffer device (hereinafter referred to as SQUID) is known as a high-sensitivity magnetic measurement device. This SQUID is a device using the quantization of magnetic flux in a superconducting loop, and is a highly sensitive element in which a superconducting loop is formed by two Josephson elements and a superconducting wiring. However, there is a substantial problem that the SQUID pickup loop must be maintained at a cooled temperature.

  As a prior art document of the present invention, a magnetic particle detector using an induction sensor coil has been proposed (see, for example, Patent Document 1). The method described in Patent Document 1 detects a magnetic substance with high sensitivity by generating a strong magnetic field at a high frequency of about several hundred kHz.

  An immunoassay method using an antigen-antibody reaction using SQUID is disclosed in Patent Document 2. According to the method described in Patent Document 2, a magnetic substance ultrafine particle is labeled on one antigen or antibody to form a magnetic substance label, and this magnetic substance label and specimen are reacted with each other by antigen-antibody reaction. Are separated and removed, and the magnetization of the specimen thereafter is measured by SQUID.

US Pat. No. 6,046,585 JP-A-63-90765 August 2003 Quantum Design Technical Documents

  However, the above-mentioned Patent Document 1 has a problem of heat generation and coil destruction due to the influence of coil inductance when generating a strong magnetic field at a high frequency. For this reason, in Patent Document 1, an annular core type electromagnet having an extremely narrow gap is used to generate a high-frequency strong magnetic field in the gap, and a sample to be measured is placed in the gap for measurement. .

  That is, this means that the thickness of the sample to be measured is greatly limited and the measurement versatility is low. The size of the gap is 0.43 mm in Non-Patent Document 1, and the thickness of the sample to be measured is necessarily limited to 0.43 mm or less. It is clear that the apparatus has the problem that the thickness of the sample to be measured is greatly limited and the measurement versatility is low.

  The present invention has been made in view of such problems, and the object of the present invention is to provide a high degree of sensitivity and a high degree of freedom with respect to the thickness of a sample to be measured in a test substance analysis using magnetic particles. It is to provide a highly versatile test substance analyzing apparatus and quantification method.

The present invention has been made in order to achieve such an object. The invention according to claim 1 is a first frequency generator that generates a magnetic field having a low frequency of 1 Hz or more and 1 kHz or less and an intensity of 50 Gauss or more. a magnetic field generating means, and a second magnetic field generating means for generating a following high frequency and a the strength 0.1 10 gauss magnetic field in gauss 10MHz at 100kHz or more, said first magnetic field generating means and the second magnetism of consists magnetic detection coil disposed in a magnetic field from the magnetic field generating means, arranged on the magnetic detection coil to detect the magnetic particles of the complex of the analyte is specifically bound magnetic particles Detecting means, and the second magnetic field generating means and the magnetic detection coil are integrally formed on a printed board, and the magnetic detection means detects the amount of change in magnetic flux of the composite, thereby detecting the object. Test substance It is characterized by quantifying.

According to a second aspect of the present invention, in the first aspect of the present invention, the first magnetic field generation means is a ferrite annular core coil or a Helmholtz coil, and the second magnetic field generation means and the magnetic detection coil Are printed coils made of copper wire .

The invention according to claim 3 is the invention according to claim 1 or 2, wherein the magnetic detection coils are connected in a differential form with two of the same shape and opposite winding directions. It is characterized by.

  The invention according to claim 4 is the invention according to claim 1, 2 or 3, wherein the specific binding is a binding between substances having specific affinity, and an antigen, an antibody, a sugar And lectins, nucleotide chains and complementary nucleotide chains, ligands and receptors, and the like.

The invention according to claim 5 is a first magnetic field generating means for generating a magnetic field having a low frequency of 1 Hz or more and 100 Hz or less and an intensity of 50 Gauss or more, and a high frequency of 1 MHz or more and 10 MHz or less. A second magnetic field generating means for generating a magnetic field with an intensity of 0.1 gauss to 10 gauss and a magnetic detection coil disposed in the magnetic field from the magnetic field generating means, the second magnetic field generating means and the And a magnetic detection means for detecting the amount of magnetic particles of a composite of magnetic particles to which a test substance is specifically bound , the magnetic detection coil being formed integrally with the printed board, and disposed on the magnetic detection coil. a test substance quantification method using the test substance analyzer having a placing said complex of the analyte is specifically bound magnetic particles on the magnetic detection coil, said first From the magnetic field A step of simultaneously generating two types of frequencies by the raw means and the second magnetic field generation means, and a magnetic particle complex in which the test substance is specifically bound in the magnetic field of the magnetic field generation means. And detecting the amount of magnetic flux change of the complex generated by the magnetic detection coil to quantify the test substance.

  Further, the invention according to claim 6 is the invention according to claim 5, wherein, when the test substance is an antigen, the step of arranging the test substance comprises: A complex of magnetic particles that are specifically bound is immobilized by an antigen-antibody reaction, and the entire base material is disposed on the magnetic detection coil.

  The invention according to claim 7 is the invention according to claim 5, wherein, when the test substance is an antigen, the placing step directly binds the antibody to the surface of the magnetic detection coil. Features.

  As a result of intensive studies to solve the above-mentioned problems, the present inventor applied two types of magnetic fields, that is, a low-frequency magnetic field of 1 kHz or less and a high-frequency magnetic field of 100 kHz or more to the sample to be measured. As a result of focusing on the detection using the nonlinearity of the permeability change of particles, we have completed a highly versatile analyte analyzer and quantitative method with high sensitivity and a high degree of freedom for the thickness of the sample to be measured. It is what led to it.

According to the present invention, the first magnetic field generating means for generating a magnetic field having a low frequency of 1 Hz or more and 1 kHz or less and an intensity of 50 Gauss or more, and a high frequency of 100 kHz or more and 10 MHz or less and an intensity of 0.1 Gauss. more than 10 and the second magnetic field generating means for generating a gauss magnetic field, made from a magnetic detection coil disposed in a magnetic field from said first magnetic field generating means of the magnetic field generating means and said second, magnetic detection A magnetic detection means for detecting a magnetic particle amount of a composite of magnetic particles specifically bound to a test substance disposed on the coil, wherein the second magnetic field generation means and the magnetic detection coil are printed. Since the test substance is quantified by being integrally formed on the board and detecting the amount of magnetic flux change of the complex by the magnetic detection means, the thickness of the sample to be measured is high. versus Thus, it is possible to realize a highly versatile test substance analyzer and quantitative method having a high degree of freedom.

Embodiments of the present invention will be described below with reference to the drawings.
FIG. 1 is a block diagram for explaining an embodiment of a test substance analyzing apparatus according to the present invention. In the figure, reference numeral 1 is a first magnetic field generating source for generating a low frequency magnetic field, and 2 is a high frequency by an exciting coil or the like. The second magnetic field generation sources 3a and 3b for generating the magnetic field indicate two magnetic detection coils. FIG. 2 is a diagram showing the second magnetic field generation source 2 and the two magnetic detection coils 3a and 3b in FIG. 1. FIG. 3 shows that a test substance is provided on the magnetic detection coils 3a and 3b in FIG. It is the figure which has arranged the plastic resin 4. Reference numeral 5 denotes a lead, A denotes the length of the exciting coil 2 in the X direction, 12 mm, and B denotes the inner diameter of the magnetic detection coils 3a and 3b, which is 2.8 mm.

  The test substance analyzer of the present invention includes a first magnetic field generation source (low frequency magnetic field generation) 1 and a second magnetic field generation source (high frequency magnetic field generation) 2 that simultaneously generate two types of frequencies, and the generation of this magnetic field. The magnetic detection coils 3a and 3b are arranged in the magnetic field from the sources 1 and 2, and the magnetic particle complex to which the test substance is specifically bound is arranged on the magnetic detection coils 3a and 3b. A magnetic detection unit for detecting the amount of magnetic particles, and the magnetic detection unit detects the amount of magnetic flux change of the complex, thereby quantifying the test substance.

  As described above, the test substance analyzer of the present invention includes the magnetic field generation sources 1 and 2 that can generate two types of frequencies simultaneously, and the magnetic detection coils 3a and 3b that detect a change in magnetization. The magnetic field frequency by the magnetic field generation source 1 of the low frequency magnetic field in the invention is 1 Hz or more and 1 kHz or less, and preferably 1 Hz or more and 100 Hz or less from the viewpoint of heat generation due to the inductance of the coil.

  The magnetic field intensity of the magnetic field source 1 must be at least 50 gauss, preferably 100 gauss or more, more preferably 200 gauss because it is necessary to apply a magnetic field intensity that causes a nonlinear effect of the magnetic particle permeability to the magnetic particles. That's it.

  Moreover, the magnetic field frequency by the magnetic field generation source 2 of a high frequency magnetic field is 100 kHz or more and 10 MHz or less, and 1 MHz or more and 10 MHz or less are preferable from the point of detection sensitivity. The magnetic field strength is preferably larger from the viewpoint of sensitivity, but it is preferably 0.1 gauss or more and 10 gauss or less in consideration of heat generation and coil destruction due to coil inductance.

  Examples of the magnetic field generating source 1 for generating a low frequency magnetic field include, but are not limited to, a ferrite annular core coil and a Helmholtz coil. Examples of the magnetic field generation source 2 for generating a high-frequency magnetic field include, but are not limited to, a printed coil made of a copper wire. Furthermore, the magnetism detection coil is configured to detect a change in magnetization from the induced electromotive force change generated in the magnetism detection coil, and detects a magnetic field change from the induced electromotive force generated by the magnetic flux change that passes through the magnetism detection coil. It is. The magnetic detection coil may be, for example, a printed coil made of copper wire, and may have a polymer film on the printed coil.

  The shape of the magnetic detection coil may be any shape, and may be circular or square, but is not limited thereto. It is also possible to use a plurality of detection coils. In this case, it is also possible to use them connected in a differential manner.

  The present invention is a test substance analyzing apparatus provided with magnetic field generating sources 1 and 2 capable of simultaneously generating two kinds of frequencies as described above, and magnetic detection coils 3a and 3b. It can be used to quantify the test substance.

  Specifically, a magnetic detection coil detects the amount of magnetic flux change caused by placing a complex of magnetic particles, to which a test substance is specifically bound, in a magnetic detection unit of the test substance analyzer. This is a method for quantifying a test substance for quantifying a substance.

  Here, specific binding refers to binding between substances having specific affinity. Examples of specific binding include antigen and antibody, sugar and lectin, nucleotide chain and complementary nucleotide chain, Examples include binding of a ligand and a receptor. For example, when the test substance is an antigen, the object to be analyzed in the present invention is to measure the specific binding between the antibody immobilized on the surface of the magnetic particles and the antigen of the test substance.

  The magnetic particles used in the present invention may be any particles that can be magnetized when a magnetic field is applied from the outside, and examples include particles containing a ferromagnetic material.

  As a ferromagnetic material, for example, a ferromagnetic metal such as iron, cobalt, nickel, an alloy containing this ferromagnetic metal, a material containing a ferromagnetic metal or an alloy containing a ferromagnetic metal in a non-magnetic material, a ferromagnetic metal Examples thereof include those containing a non-magnetic substance in an inside or an alloy containing a ferromagnetic metal.

  The particle diameter of the magnetic particles is preferably not too large from the viewpoint of reaction efficiency, and preferably not too small from the viewpoint of measurement sensitivity. Therefore, the average particle diameter of the magnetic particles of the present invention is 0.001 μm to 10 μm, preferably 0.01 μm to 5 μm, more preferably 0.1 μm to 3 μm. The average particle diameter can be measured by a known method such as a dynamic light scattering method or an electron microscope.

  This magnetic particle is generally called a superparamagnetic substance, and it is particularly preferable that the magnetic particle has a property of being magnetized while a magnet is applied from the outside and demagnetizing rapidly by blocking the magnet from the outside. Examples of such magnetic particles include Dynabeads (registered trademark) M-450, Dynabeads (registered trademark) M-280, Dynabeads (registered trademark) Myone (trademark), Merck Chime, S. A. , S Estapor (registered trademark) M1-070-40, Estapor (registered trademark) M1-070-60, Estapor (registered trademark) M1-030-40, Seradyn Sera-mag (trademark), etc. . As the magnetic particles, those that have been subjected to an appropriate surface treatment, those having an appropriate functional group, and the like are selected according to the type of immunoreactive substance to be applied.

  In addition, the magnetic particles may be preliminarily provided with a substance that specifically binds to the test substance, for example, when the test substance is an antigen, the antibody is given by chemical adsorption or physical adsorption by a known method. In many cases, a complex of magnetic particles formed by specifically binding a test substance to such magnetic particles is measured by the test substance analyzer of the present invention.

  In FIG. 1, although the Helmholtz coil is used as the magnetic field generation source 1 for generating the low frequency magnetic field, it is not limited to this. In FIG. 2, the magnetic field generation source 2 for generating a high-frequency magnetic field and the magnetic detection coils 3a and 3b are printed coils in the same plane, but the magnetic field generation source 2 includes the magnetic detection coils 3a and 3b. If it is a form, it is not limited to these. Further, in FIG. 2, the magnetic detection coils are configured such that the two magnetic detection coils 3a and 3b are connected in a differential manner, but are not limited thereto.

  In the analysis of the test substance, a magnetic particle complex in which the test substance is specifically bound is arranged on the magnetic detection coils 3a and 3b in the arrangement shown in FIG. The test substance can be quantified.

  Further, as shown in FIG. 2, when two magnetic detection coils 3a and 3b are used, a complex of magnetic particles to which a test substance is specifically bound is disposed only on one of them, and the other is for reference. Used as a magnetic sensor.

  In the present invention, a method for analyzing a composite of magnetic particles, to which a test substance is specifically bound, placed in a magnetic detection unit will be specifically described in the case of using a test substance as an antigen.

  There are the following two methods for arranging a composite of magnetic particles in which a test substance is specifically bound to the magnetic detection unit.

  First, the first method is a method in which a complex of magnetic particles in which a test substance is specifically bound to a substrate having an antibody on its surface is immobilized by an antigen-antibody reaction and arranged together with the substrate. The base material used here may be any material that can bind an antibody such as polyacrylonitrile to the surface by a known method.

  The analysis method using a magnetic particle complex in which a test substance is specifically bound to the base material is used for the analysis. Since it can be analyzed, it is effective in that a large number of analyte measurement samples can be analyzed quickly.

  In the second method, an antibody is directly bound to the surface of a magnetic detection coil, and a complex of magnetic particles to which a test substance is specifically bound is immobilized by an antigen-antibody reaction for analysis. Such a method is effective in that it is possible to generate a higher magnetic field in that it is not necessary to dispose a substrate having a thickness as compared with the first method described above.

  Next, the magnetic detection method in the test substance analyzer of the present invention will be described below. Hereinafter, although the case where two magnetic detection coils 3a and 3b are used will be described, the number of magnetic detection coils is not limited to two.

  As shown in FIG. 2, the magnetic detection coils 3 a and 3 b are connected in a differential manner, and the two magnetic detection coils 3 a and 3 b have the same shape and the winding directions are opposite to each other. . When detecting magnetic particles, as shown in FIG. 1, a magnetic field is simultaneously applied from the magnetic field generation source 1 and the magnetic field generation source 2 to the magnetic detection coils 3a and 3b. Since the magnetic fields from the magnetic field generation source 1 and the magnetic field generation source 2 are alternating magnetic fields that vary with frequency, the magnetic flux passing through the magnetic detection coils 3a and 3b changes with time, and is induced in each of the magnetic detection coils 3a and 3b. An electromotive force is generated. Since the induced electromotive force is proportional to the applied magnetic field strength and the applied magnetic field frequency, the induced electromotive force detected by the magnetic detection coils 3a and 3b in the present invention is due to the change in magnetic flux of the test substance by the magnetic field generation source 2.

  In FIG. 2, the induced electromotive force generated in each of the magnetic detection coils 3a and 3b has the same strength and opposite phase due to the coupling configuration of the two magnetic detection coils 3a and 3b. As shown in FIG. 2, since the two sensor coils are connected in a differential manner, the signal output is canceled in the absence of magnetic particles.

Next, a method for detecting magnetic particles to which a test substance is specifically bound will be described.
FIG. 4 is a diagram showing a magnetization curve of magnetic particles to which a test substance is specifically bound. As shown in FIG. 4, the magnetic permeability of the magnetic particles has a nonlinear response to the externally applied magnetic field strength. As shown in the present invention, when the magnetic field generation source 1 applies a magnetic field having a magnetic particle permeability that exhibits a nonlinear effect to the magnetic particles, the point 5 at which the magnitude of the magnetic field generated by the magnetic field generation source 1 is maximized. In this point, the change in magnetic permeability of the magnetic particles is small.

  On the other hand, at the point 6 where the magnitude of the magnetic field generated by the magnetic field generation source 1 is minimum, the magnetic particle permeability change is in a linear region, and the magnetic particle permeability change is maximum. When a high frequency alternating magnetic field is applied to the magnetic particles by the magnetic field generation source 2 while the alternating magnetic field is applied to the magnetic particles by the magnetic field generation source 1, the change in the magnetic permeability of the magnetic particles by the magnetic field generation source 2 at point 5 is minimized. The induced electromotive force generated in the magnetic detection coils 3a and 3b is also minimized.

  On the other hand, the change in the magnetic permeability of the magnetic particles by the magnetic field generating source 2 at the point 6 is maximized, and the induced electromotive force generated in the magnetic detection coils 3a and 3b is maximized. By measuring the difference between the induced electromotive forces generated at point 5 and point 6, the amount of magnetic particles as the sample to be measured can be measured.

  FIG. 5 is a diagram showing a signal output waveform of magnetic particles with respect to a magnetic source. From FIG. 5, the change in the induced electromotive force due to the magnetic particles with respect to the magnetic field generation source 2 is minimized at the point where the magnitude of the magnetic field by the magnetic field generation source 1 is maximized, and the magnitude of the magnetic field by the magnetic field generation source 1 is minimized. It can be seen that the change in the induced electromotive force due to the magnetic particles with respect to the magnetic field generation source 2 becomes the maximum.

The test substance analyzer and the quantification method of the present invention have been described above.
Next, a method for detecting magnetic particles of a test substance using the test substance analyzer of the present invention will be described below.

<Magnetic particle detection system>
FIG. 6 is a diagram showing a magnetic detection coil in the magnetic particle detection system.
As shown in FIG. 6, as the magnetic detection coil, a printed coil having an inner diameter B of 2.8 mm and a winding number of 5 is used. In the present invention, two magnetic detection coils 3a and 3b are connected in a differential manner to form magnetic particles. Detection was performed.

  As shown in FIG. 6, the magnetic field generation source 2 has a configuration including magnetic detection coils 3 a and 3 b, and a printed coil having an inner diameter Ax × Ay (12 mm × 7 mm) and 5 windings was used. An alternating magnetic field was generated in the magnetic field generation source 2 using an alternating current. Magnetic particles were detected by setting the magnetic field frequency of the magnetic field generation source 2 to 2.24 MHz and the magnetic field intensity to 1 gauss.

  For the magnetic field generation source 1, an air-core Helmholtz coil was used. An alternating magnetic field was generated in the gap portion of the magnetic field generation source 1 using an alternating current. Magnetic particles were detected by setting the magnetic field frequency of the magnetic field generation source 1 to 63 Hz and the magnetic field intensity to 150 gauss.

  FIG. 7 is a block diagram showing a signal processing system in the test substance analyzer of the present invention. In the figure, reference numerals 7 and 17 are frequency generators, 8 is a power splitter, 9, 12, 15, and 18 are amplifiers, A printed board, 13 is a frequency mixer, 16 is a lock-in amplifier, and 20 is a sample to be measured.

  The alternating current from the frequency generator 7 is divided into two by the power splitter 8, and one is input to the frequency mixer 13. The other was amplified by an amplifier 9 and input to the magnetic field generation source 2 to generate an alternating magnetic field by the magnetic field generation source 2, and a magnetic field was applied to the magnetic detection coils 3a and 3b. In the present invention, the printed board 10 in which the magnetic field generation source 2 and the magnetic detection coils 3a and 3b are integrated is used. The printed board 10 is provided with a sample 20 to be measured, and the sample 20 to be measured functions as a composite of magnetic particles to which a test substance is specifically bound.

  A signal obtained from the magnetic detection coils 3 a and 3 b was taken out from the wiring 14, the signal was amplified by the signal amplifier 12, and the amplified signal was input to the frequency mixer 13. The frequency mixer 13 frequency-mixes the input signal and the amplified signal from the frequency generator 7, and the obtained magnetic signal is amplified by the signal amplifier 15 and input to the lock-in amplifier 16.

  On the other hand, the alternating current from the frequency generator 17 is input to the magnetic field generation source 1 by the amplifier 18 to generate an alternating magnetic field by the magnetic field generation source 1 and simultaneously apply the magnetic field to the magnetic detection coils 3a and 3b, and at the same time, a lock-in amplifier. 16 was input as a reference signal.

  The magnetic signal was detected by detecting the reference signal and the magnetic signal from the frequency generator 17 using a phase detection technique which is a well-known technique using the lock-in amplifier 16.

<Preparation of sample to be measured>
Magnetic particles were immobilized on the bottom surface of the sample 20 to be measured made of the plastic resin 4 having a thickness of about 6 mm using an antigen-antibody reaction. The magnetic particles were fixed in a circular shape having a diameter of about 2 mm. In addition, Dynabeads Myone was used as the magnetic particles.

  FIG. 8 is a configuration diagram of a test substance analyzer for explaining a method for detecting magnetic particles, in which 19 is a fixing unit for magnetic particles, 20 is a sample to be measured, and the other functions are the same as those in FIG. Elements are given the same reference numerals.

  As shown in FIG. 8, a sample 20 to be measured is provided with a fixed part 19 of a composite of magnetic particles to which a test substance is specifically bound, and this fixed part 19 is connected to one of the magnetic detection coils 3a, The measurement was carried out by placing it on 3b.

  Table 1 shows the results of magnetic particle detection using the above-described magnetic particle detection system and the sample to be measured.

  As shown in Table 1, the test substance analysis apparatus of the present invention can detect magnetic particles with high sensitivity with a high degree of freedom with respect to the thickness of the sample to be measured.

It is a block diagram for demonstrating one Example of the to-be-tested substance analyzer which concerns on this invention. It is a figure which shows the 2nd magnetic field generation source in FIG. 1, and two magnetic detection coils. It is the figure which has arrange | positioned the plastic resin which provided the to-be-tested substance on the magnetic detection coil in FIG. It is a figure which shows the magnetization curve of the magnetic particle which the test substance specifically couple | bonded. It is the figure which showed the signal output waveform of the magnetic particle with respect to a magnetic generation source. It is a figure which shows the magnetic detection coil in the detection system of a magnetic particle. It is a block diagram which shows the signal processing system in the test substance analyzer of this invention. It is a block diagram of the test substance analyzer for demonstrating the detection method of a magnetic particle.

Explanation of symbols

DESCRIPTION OF SYMBOLS 1 1st magnetic field generation source 2 2nd magnetic field generation sources 3a and 3b Two magnetic detection coils 4 Plastic resin which provided the test substance 5 Leads 7 and 17 Frequency generator 8 Power splitter 9, 12, 15, 18 Amplifier 10 Print board 13 Frequency mixer 14 Wiring 16 Lock-in amplifier 19 Magnetic particle fixing part 20 Sample to be measured

Claims (7)

First magnetic field generating means for generating a magnetic field having a low frequency of 1 Hz or more and 1 kHz or less and an intensity of 50 Gauss or more;
A second magnetic field generating means for generating a magnetic field having a high frequency of 100 kHz or more and 10 MHz or less and an intensity of 0.1 Gauss or more and 10 Gauss or less ;
Consists magnetic detection coil disposed in a magnetic field from said first magnetic field generating means of the magnetic field generating means and said second, magnetic disposed on the detection coil, the magnetic particles analyte bound specifically Magnetic detection means for detecting the amount of magnetic particles of the composite of
The second magnetic field generating means and the magnetic detection coil are integrally formed on a printed board , and the test substance is quantified by detecting the magnetic flux change amount of the complex by the magnetic detection means. A test substance analyzer characterized by the above. 2. The first magnetic field generating means is a ferrite annular core coil or a Helmholtz coil, and the second magnetic field generating means and the magnetic detection coil are printed coils made of copper wire. Test substance analysis equipment. The test substance analysis apparatus according to claim 1 or 2, wherein the magnetic detection coils are connected in a differential manner with two having the same shape and opposite winding directions .   The specific binding is a binding between substances having specific affinity, and includes an antigen and an antibody, a sugar and a lectin, a nucleotide chain and a complementary nucleotide chain, a ligand and a receptor, and the like. The test substance analyzer according to claim 1, 2 or 3. A first magnetic field generating means for generating a magnetic field having a low frequency of 1 Hz or more and 100 Hz or less and an intensity of 50 Gauss or more, and a high frequency of 1 MHz or more and 10 MHz or less and an intensity of 0.1 Gauss or more and 10 Gauss or less. A second magnetic field generating means for generating a magnetic field and a magnetic detection coil arranged in the magnetic field from the magnetic field generating means, and the second magnetic field generating means and the magnetic detection coil are integrally formed on a printed board. And using a test substance analyzing apparatus provided on the magnetic detection coil and provided with a magnetic detection means for detecting a magnetic particle amount of a complex of magnetic particles to which the test substance is specifically bound. A test substance quantification method comprising:
Arranging a complex of magnetic particles to which the test substance is specifically bound on the magnetic detection coil;
Generating two types of frequencies simultaneously by the first magnetic field generating means and the second magnetic field generating means;
A magnetic flux change amount of the complex generated by arranging a complex of magnetic particles to which the test substance is specifically bound in the magnetic field of the magnetic field generating means is detected by the magnetic detection coil, and the test substance is detected. A method for quantifying a test substance, comprising the step of:   In the case where the test substance is an antigen, the arranging step immobilizes a complex of magnetic particles to which the test substance is specifically bound to a substrate having an antibody on the surface by an antigen-antibody reaction, The test substance quantification method according to claim 5, wherein the whole substrate is disposed on the magnetic detection coil.   6. The method for quantifying a test substance according to claim 5, wherein, when the test substance is an antigen, the arranging step directly binds an antibody to the surface of the magnetic detection coil.

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