JP2001083154A - Disease marker substance simplified compact detection device - Google Patents

Abstract

PROBLEM TO BE SOLVED: To obtain a disease marker simplified compact detection device that can accurately and easily detect the amount of disease marker substance even in a house of a patient as well as a small-scale hospital and is compact, can be handled easily, and is suitable for daily monitoring. SOLUTION: The disease marker substance simplified compact detection device is provided with a means A for capturing a disease marker substance by the latex agglutination reaction, a means B for allowing the captured disease marker substance to a crystal vibrator, and a means C for oscillating the crystal oscillator where the disease marker substance has adhered and for measuring and displaying the frequency change in combination. Then, in the means C, a means D for oscillating a crystal vibrator where the disease marker substance has adhered to, a means E for measuring a frequency change being generated by the oscillation of the crystal oscillator, and a means F for displaying a measurement frequency are incorporated in one piece.

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001]

BACKGROUND OF THE INVENTION 1. Field of the Invention The present invention relates to a simple and compact detection device which can easily detect the amount of a disease marker substance appearing in a living body when a disease is sick, even in a general home or a small hospital.

[Prior art]

[0002] As is well known, the amount of disease marker substances contained in human blood, sweat, feces, urine, and saliva is an indicator of the degree of infectious disease. Analytical methods and measuring methods have been actively studied. Have been. Conventionally, such disease marker substances contained in blood, sweat, feces, urine, and saliva have been analyzed by optically detecting the agglutination of immune latex.
Since this method has an advantage that a disease marker substance to be detected can be widely selected by replacing an antibody or an antigen immobilized on latex, various immunolatex kits for detection are commercially available. For example, immunolatex kits for automatic analyzers include hepatitis B virus diagnosis (HBs antigen / antibody), hepatitis C virus diagnosis (HCV antibody), C-reactive protein test (CRP),
Rheumatism diagnosis (RA), streptococcal infection diagnosis (ASO), syphilis diagnosis (STS), AIDS diagnosis (HIV), and tumor marker diagnosis include carcinoembryonic antigen (CE)
A), α-fetoprotein (AFP), carbohydrate antig
en 19-9 (CA19-9), ferritin, prostatic aci
d phosphatase (PAP), tissue polypeptide antige
n (TPA) and others, and allergic disease diagnostic (IgE) reagents and the like are commercially available. However, these immunolatex kits require the use of expensive and large-sized automatic analyzers for blood and the like, so that they are not widely used in small hospitals and are not suitable for monitoring the health status of ordinary households at home.

In Japan, the population is rapidly aging, and it is expected that one fourth of the population will become elderly in 2020. The large increase in the number of persons with disabilities due to the aging of the population has become a major social problem, and there is an urgent need for the development of medical and welfare equipment to support the social participation of persons with disabilities. For example, pneumonia is the leading cause of death for elderly people in Japan, and early detection of pneumonia becomes possible by measuring the amount of CRP in the blood. The patient monitors the amount of CRP in his blood on a daily basis with a sensor, contacts a doctor on standby at a hospital or medical center via telephone or computer network, and receives a diagnosis of the degree of infection. Health management at home can be performed satisfactorily, and social participation support is also possible. Therefore, there is an urgent need to develop a detection device capable of easily measuring the amount of a disease marker substance at home of a sick person and a small and portable disease sensor system for monitoring.

In order to deal with such a problem, first,
The present inventors have proposed a detection method in which a change in the amount of a proteinaceous marker substance in serum is read by a change in the frequency of a quartz oscillator (ACS Symposium Series 657, 1997, chapter 15,
pp.185-196). However, according to subsequent studies by the present inventors, the apparatus used for this detection method has the advantage that an expensive and complicated large-sized automatic analyzer such as blood does not need to be used as in a conventional immunolatex kit. However, it was found that there were the following problems. This detection device is a device (oscillation circuit) that oscillates a quartz oscillator to which a disease marker substance is attached individually and separately, and a measuring device (frequency meter) of a frequency change generated by oscillation of the quartz oscillator. , A configuration in which each measurement device such as a display device (display unit) of the measured frequency, a stabilized power supply, and a computer is connected by a device such as a BNC cable or a GP-IB cable. Since the device also has a function as an antenna, such a device is easily affected by induction noise from a power source and external noise, and there is a possibility that an error or noise may occur in obtained measurement data. It becomes difficult to design an oscillation circuit for stably oscillating the quartz oscillator in the latex agglutination reactor, and thus lacks stability as a detection device. A device that oscillates a crystal oscillator to which a disease marker substance is attached (oscillation circuit), a device that measures the change in frequency oscillated by oscillation of the crystal oscillator (frequency meter), a display device that displays the measured frequency (display unit), and a stable device The measurement equipment such as a computerized power supply and a computer is arranged in a disparate manner, and a large number of connecting equipment such as BNC cables and GP-IB cables for connecting these are required. And cannot be installed in ordinary households. Since it requires skill in connection and handling of the device, it is not suitable for use in general homes and small clinics.

[0005]

SUMMARY OF THE INVENTION The present invention overcomes the above-mentioned drawbacks of the prior art, and enables accurate and convenient detection of the amount of a disease marker substance in a small clinic as well as at home of a sick person. It is an object of the present invention to provide a simple and compact disease marker substance detection device which is easy to handle and suitable for daily monitoring.

[0006]

Means for Solving the Problems As a result of intensive studies to achieve the above object, the present inventor has found that a device (oscillation circuit) for oscillating a crystal oscillator having a disease marker substance attached thereto and an oscillation of the crystal oscillator are provided. Measuring device (frequency meter) for generated frequency change and display device (display unit) for measured frequency
It has been found that it is effective for the above-mentioned problem to incorporate and incorporate into a housing, and have completed the present invention.
That is, according to the present invention, first, (A) means for capturing a disease marker substance by latex agglutination reaction;
(B) means for attaching the captured disease marker substance to the quartz oscillator, and (C) means for oscillating the quartz oscillator to which the disease marker substance is attached and measuring and displaying a change in frequency thereof. Means for oscillating the crystal oscillator to which the disease marker substance is attached and measuring and displaying the frequency change of the crystal oscillator to which the disease marker substance is attached in the case (C) Means for oscillating the vibrator,
(E) a means for measuring a change in oscillation frequency generated by oscillation of the quartz oscillator and (F) a means for displaying the change in frequency, which is incorporated and integrated into a simple disease marker substance. A small detection device is provided. Secondly, in the first device, there is provided a simple and compact disease marker substance detecting device having a portable shape. Thirdly, in the first or second device, a disease marker substance further comprising a means for issuing a caution signal when the detected value displayed on the display unit indicates a value equal to or more than a predetermined value indicating disease. A simple and compact detection device is provided. Fourth, a simple method for detecting a disease marker substance using any one of the first to third simple small-sized detection devices is provided.

[0007]

BEST MODE FOR CARRYING OUT THE INVENTION A simple and compact detection device for a disease marker substance of the present invention will be described with reference to the drawings. In FIG. 1, (A) is a means for capturing a disease marker substance by latex agglutination (latex agglutination reaction vessel), (B)
Means for attaching the captured disease marker substance to the crystal oscillator (crystal oscillator); and (C) means for oscillating the crystal oscillator to which the disease marker substance is attached, and measuring and displaying a change in oscillation frequency thereof. (Detector main body). The detector main body includes means (oscillation circuit) for oscillating a crystal oscillator (D) having a disease marker substance attached thereto in a housing and (E) oscillation of the crystal oscillator. Means (frequency meter) for measuring the generated oscillation frequency change and (F) means for displaying the measured frequency (oscillation frequency display) are integrated and integrated. In FIG. 1, the unit is mm.

Specifically, the means (detector main body) for oscillating the crystal oscillator to which the disease marker substance (C) is attached and measuring and displaying a change in the frequency thereof is provided in a metal housing.
(D) Oscillation circuit of a crystal oscillator devised to stably oscillate in an immune latex dispersion solution.
(E) As a frequency meter, a frequency measuring kit (PIC Kit ver. 2.00) of Akizuki Tsusho and (F) a liquid crystal display unit or the like are incorporated and appropriately integrated by a coupling means.

The quartz resonator used in the present invention exhibits a larger oscillation frequency response during the immune latex agglutination reaction as its fundamental frequency is higher. However, it is preferable to use about 9 MHz because the noise increases and the frequency stability decreases.
Also, in order to stably oscillate the crystal oscillator in the immune latex solution, only one side of the crystal oscillator may be in contact with the solution. Adhering method is adopted. Silver or gold is preferable as the electrode material of the crystal unit, and silver is preferable in terms of frequency response and inexpensiveness, but high frequency stability in a solution and storage in air before use. Gold is more preferred because it is resistant to oxidation of the electrode surface.

There are two typical examples of the small and simple disease marker substance detecting apparatus of the present invention. One of them is to use an LED display as a measurement frequency display unit, and is driven by a 9 V AC adapter because of high power consumption. The other one uses a liquid crystal display for the measurement frequency display unit, and uses a small amount of power and is driven by a dry battery (one of 9V and 006P is used). The dry batteries can be replaced with rechargeable batteries of the same capacity. By the way, the size and weight of these two devices are 9x20x10cm and 1k
g (including AC adapter), the latter is 11 × 13 × 3c
m is 0.5 kg (including dry batteries), and both devices are extremely small in shape and light in weight.

As described above, in the conventional specific disease marker substance measuring device, measuring devices such as a frequency meter, an oscillation circuit, a stabilized power supply, and a computer are individually and randomly arranged, and a BNC cable and a GP are used. -It is necessary to connect the devices by IB cables, and these cables become antennas and are susceptible to external noise such as induction noise from a power supply, which greatly affects measurement data. In this case, since each device is connected and integrated without using these cables, noise is prevented from being mixed into the measurement data, and the accuracy of the measurement data is improved.

Also, in the conventional method for measuring a specific disease marker substance, it is difficult to design an oscillation circuit for stably oscillating the quartz oscillator in the latex agglutination reactor, and the surface lacks stability as a detection sensor. However, in the present invention, the oscillation circuit was integrated with the crystal unit and the battery, and was sealed in a metal housing and a specific circuit constant was selected. As a result, it is possible to obtain a detection device having excellent stability.

Further, the conventional disease marker substance detecting device is an attached device for oscillating a quartz oscillator (oscillator circuit), a measuring device for measuring the frequency generated by the oscillation of the quartz oscillator (frequency meter), and the measured frequency. Display devices (display units), stabilizing power supplies, computers, and other measuring instruments are arranged separately, and a large number of BNC cables and GP-IB cables are required to connect them. However, there is a disadvantage that the apparatus becomes large and cannot be installed in a small clinic or a general home. However, in the present invention, the crystal oscillator to which the disease marker substance of (C) is attached oscillates and its frequency change occurs. Means for measuring and displaying is generated by means for oscillating a crystal oscillator having a disease marker substance attached in a housing and (E) oscillation of the crystal oscillator. Since the means for displaying the unit and (F) the measurement frequency to measure a frequency change has a configuration in which integrated are integrated, BNC cable and GP-IB
It eliminates the need for connecting devices such as cables, and makes it possible to reduce the size of the device to about 11 × 13 × 3 cm and the total weight to about 0.5 kg.
Its size and weight can be significantly reduced compared to 120 × 100 cm and 20 kg. As a precautionary measure, FIG. 3 shows a comparison of the shape of the conventional device and the device of the present invention in the disease marker substance detecting device. The unit is mm.

The conventional apparatus for detecting a disease marker substance requires skill in connection, handling, and the like. However, the apparatus of the present invention can perform the connection and measurement operations with a very simple operation. It can be easily used in small clinics.

The apparatus of the present invention has the above-mentioned configuration, but preferably further includes a means for issuing a caution signal when the detected value displayed on the display unit is equal to or higher than a predetermined value indicating disease. According to the device equipped with such a means, it is possible to easily detect the extent of the specific infection and the presence or absence of the infection, and to send the measurement data directly to a medical center or a doctor at a hospital by telephone or computer network. ,
It has the advantage that it can be used for diagnosis of remote infectious diseases at home of a sick person and as a monitoring method of the health condition of the sick person.

The disease marker substances to be monitored in the present invention include the aforementioned protein disease marker substances such as HBs, HCV, CRP, RA, ASO, STS and STS.
HIV, CEA, AFP, CA1 for tumor marker diagnosis
9-9, ferritin, PAP, TPA, etc.
These immune latexes and standard serums, such as gE reagents, are commercially available as kits for automatic blood analyzers and can be used in the present detection sensor system. In the case of measurement of other disease marker substances, it is possible to measure the amount by preparing an antibody of the molecule to be measured and immobilizing it on latex for latex agglutination.

In order to detect a disease marker substance in a solution using the apparatus of the present invention, first, an immunolatex solution is filled in a latex agglutination reaction vessel (reaction cell), and then one side of a quartz oscillator is immersed in the reaction cell. After stabilizing the oscillation frequency, a solution containing the antigen to be measured is added. The frequency 60 minutes after the completion of the aggregation of the immune latex may be read from the display, and the frequency difference before and after the addition of the antigen may be calculated.

[0018]

Now, the present invention will be described in further detail with reference to embodiments. These examples are for facilitating the understanding of the present invention, and do not limit the scope of the present invention in any way.

EXAMPLE The following measurement was carried out using a disease marker substance detecting device as shown in FIG. (B) A single-sided sealed crystal unit was replaced with (A) a measurement cell (methyl methacrylate, 1
0x10x20mm) phosphate buffer (pH
6.50, I = 10 mM) was placed in a stirring solution of 1.2 ml of CRP antibody-fixed latex and 0.1 ml of a CRP antibody-fixed latex. When the oscillation frequency became stable, CRP standard serum (CRP concentration of 4.4 mg / dl, addition volume of 2.5 to 2.5 ml) was added. 17.5 μl) was added, and the change in the oscillation frequency of the vibrator was read on the display (F) of the detector. The addition of CRP-containing standard serum rapidly reduced the oscillation frequency of the quartz oscillator to about 6
It converged to a constant value after 0 minutes. The convergence value of this oscillation frequency change was determined as the end point of the immunoagglutination reaction of the CRP antibody-fixed latex. FIG. 4 shows the CRP concentration measurement results obtained by the present detection apparatus when CRP-containing and CRP-free (CRP-free) standard serum was added. In addition, ● indicates serum containing CRP and ○ indicates CRP-free serum (average value of each n ≦ 5, fluctuation within 30%). From FIG. 4, the change in the crystal oscillation frequency occurs from around 0.01 mg / dl, and 0.06 mg / d
It can be seen that it is proportional to about l. The CRP concentration can be determined by the response of the sensor. The concentration determined as CRP-positive in the clinical test is 1 mg / dl or more. The measurement sensitivity of the present detection device is more than 100 times higher and has sufficient sensitivity for CRP measurement. Also, CR
Oscillation frequency change occurs even in serum without P,
It is clear from FIG. 4 that it is significantly smaller than the change due to CRP.

[0020]

The simple and compact detection device and method for detecting a disease marker substance according to the present invention have the following functions and effects. (1) In the conventional specific disease marker substance measuring device, measuring devices such as a frequency meter, an oscillation circuit, a stabilized power supply, and a computer are individually and randomly arranged.
Cables and GP-IB cables need to be connected between devices, and these cables become antennas and are susceptible to external noise such as induction noise from the power supply, which has had a large effect on measurement data. In the device according to the invention, since each device is connected and integrated without using these cables, noise is prevented from being mixed into the measurement data, and the accuracy of the measurement data is improved. (2) In the conventional method for measuring a specific disease marker substance, it is difficult to design an oscillation circuit for stably oscillating a quartz oscillator in a latex agglutination reactor, and there is a face that lacks stability as a detection sensor. However, in the present invention, since the oscillation circuit is integrated with the crystal oscillator and the battery, and is enclosed in a metal housing and a specific circuit constant is selected, the crystal oscillator oscillates stably even in the latex agglutination reactor. Therefore, a detection sensor having excellent stability can be obtained. (3) A conventional disease marker substance detecting device is a device that oscillates an attached crystal oscillator (oscillation circuit), a device that measures a frequency generated by oscillation of the crystal oscillator (frequency meter),
Display device (display unit) of measured frequency, stabilized power supply,
Each measuring device such as a computer is placed separately,
Moreover, since many connecting devices such as BNC cables and GP-IB cables for connecting these are required,
There was a drawback that the device became large and could not be installed in a small clinic or home, but in the present invention,
(C) means for oscillating the crystal resonator to which the disease marker substance is attached, and means for measuring and displaying the generated frequency; and (D) means for oscillating the crystal resonator to which the disease marker substance is attached in the housing. And (E) means for measuring the frequency generated by the oscillation of the crystal unit, and (F) means for displaying the measured frequency are integrated and integrated.
Eliminates the need for connecting equipment such as BNC cables and GP-IB cables, and reduces the total volume of the equipment to 11 × 13 × 3 cm
And the total weight can be reduced to about 0.5 kg, which is 200 × 120 × 100 cm, 2
The size and weight can be significantly reduced compared to 0 kg. (4) The conventional disease marker substance detection device required skill in connection and handling, but the device of the present invention can perform the connection and measurement operation with very simple operation.
It can be easily used in ordinary households and small clinics. (5) The method for detecting a disease marker substance of the present invention focuses on the fact that a change in the oscillation frequency of a quartz oscillator occurs in proportion to the amount of a disease marker substance to be measured in an immune latex agglutination reaction. Since the amount is measured by a simple and compact detection device / system that integrates an oscillation circuit and a frequency meter, the amount of the disease marker substance can be accurately and easily detected not only at small clinics but also at home for patients with illness. Moreover, the handling operation is easy and suitable for daily monitoring. (6) In the device / method for detecting a disease marker substance of the present invention, if the detected value indicates a value equal to or more than a predetermined value indicating the presence of an infectious disease, a caution signal is issued, so that the occurrence of the specific infectious disease is achieved. Diagnosis of remote infectious diseases at home and monitoring the health of infected patients by sending measurement data to doctors at medical centers and hospitals by telephone or computer network to detect the degree of infection and the presence or absence of infection Available as (7) Since the detection device of the present invention has a high CRP minimum detection sensitivity of 0.008 mg / d (CRP positive concentration of 1 mg / dl or more), for example, a crystal oscillator of coexisting protein in serum Even if non-specific adsorption to CRP occurs, CRP can be accurately detected and can be sufficiently used in the field of clinical testing. (8) The crystal oscillator type portable disease marker substance detection device of the present invention is inexpensive and easy to purchase even in ordinary households.

[Brief description of the drawings]

FIG. 1 is an explanatory diagram of a simple and compact disease marker substance detection device according to the present invention.

FIG. 2 is an explanatory diagram of an oscillation circuit used in the present invention.

FIG. 3 is a comparison diagram of the device of the present invention and a conventional device.

FIG. 4 is a graph showing the relationship between the change in the oscillation frequency of the quartz oscillator measured by the apparatus of the present invention and the CRP concentration.

Claims (4)

[Claims] 1. A means for capturing a disease marker substance by latex agglutination, (B) means for attaching the captured disease marker substance to a quartz oscillator, and (C) a crystal to which the disease marker substance is attached. Means for oscillating the vibrator and measuring and displaying the frequency change thereof, and oscillating the crystal vibrator to which the disease marker substance of (C) is attached and measuring and displaying the frequency change Means for oscillating a crystal oscillator having a disease marker substance adhered in a housing; (E) means for measuring a change in oscillation frequency generated by oscillation of the crystal oscillator; and (F). A simple and compact disease marker substance detection device, wherein a means for displaying the frequency change is incorporated and integrated. 2. The small and simple disease marker substance detecting apparatus according to claim 1, wherein the apparatus is portable. 3. A simple and compact disease marker substance according to claim 1 or 2, further comprising a means for issuing a caution signal when the detected value displayed on the display section is equal to or higher than a predetermined value indicating disease. Detection device. 4. A simple method for detecting a disease marker substance using the simple and compact detection device according to claim 1.