Collection method, inspection method, centrifugal device, and inspection system

Abstract

A simple and stable method by which samples in different forms can be acquired at once from a blood sample.　A method for collecting microorganisms that comprises: introducing a blood sample 200 into a first holding part 100 of a container 1, said container 1 being provided with the first holding part 100 having a first flow channel 103 and a second flow channel 104 of different heights on the side face, a second holding part 101 and a third holding part 102; centrifuging the blood sample 200 at a first centrifugal force to separate into a microbial solution 202 containing microorganisms and a solution 203 containing blood cells; centrifuging the blood sample 200 at a second centrifugal force greater than the first centrifugal force and moving the solution 203 through the first flow channel 103 to the second holding part 101; centrifuging the second solution 203 at a third centrifugal force to separate into a microbial solution 206 containing microorganisms and blood components 207 containing blood cells; and centrifuging the solution 203 at a fourth centrifugal force greater than the third centrifugal force and moving the microbial solution 206 through the second flow channel 104 to the third holding part 102.

Description

Technical Field

[0001] The present invention relates to a collection method, an inspection method, a container, a centrifuge, and an inspection system. More specifically, the present invention relates to a method for collecting microorganisms that separates a solution containing microorganisms such as bacteria and a solution containing blood cells from a sample containing impurities such as blood cells, a method for inspecting microorganisms, a container, a centrifuge, and an inspection system.

Background Art

[0002] Sepsis is an infectious disease with a high fatality rate, and it is important to perform appropriate treatment promptly. Usually, a blood culture test is performed to determine sepsis, and it is determined whether bacteria are present in the blood, which is a sterile sample. Generally, a smear test is then performed, the blood culture positive sample is isolated and cultured, an identification test for identifying the type of bacteria against the obtained colonies, and a sensitivity test for measuring the sensitivity of the bacteria to antibacterial drugs are performed. The above series of tests require 1 day for the blood culture test, 1 day for the isolation culture, and 1 day for the sensitivity test, so the total test time is 2 to 3 days. Therefore, it currently takes 2 to 3 days to determine whether appropriate treatment with an antibacterial drug has been implemented, and if an ineffective antibacterial drug has been administered, the fatality rate becomes extremely high.

[0003] Currently, attempts to speed up this series of infectious disease tests are being actively researched. For example, in bacterial identification tests, the conventionally used colorimetric and fluorescence methods were the gold standards. Currently, a method using mass spectrometry by MALDI-TOF (Matrix Assisted Laser Desorption / Ionization - Time Of Flight) for identification tests has been developed, and it has seen a remarkable spread from the perspectives of test speed and cost-effectiveness. Regarding susceptibility tests, the turbidimetric method has been conventionally used, but methods for highly sensitive detection of bacteria using lasers and methods for predicting susceptibility based on the growth and morphological changes of bacteria using microscopes have been developed. Thus, the tests are becoming faster. On the other hand, separation and culture still remain as a time-consuming step in the tests and have become a bottleneck in shortening the test time. Here, if it is possible to remove, for example, blood cell components and other impurities other than bacteria from a blood culture positive sample and extract only the bacteria, the separation and culture step can be omitted, leading to a shortening of the test time.

[0004] In ordinary bacterial tests, identification tests and susceptibility tests are performed as a set. Therefore, when extracting bacteria from a blood culture positive sample, it is necessary to prepare a sample suitable for both tests. If a sample suitable for both tests can be obtained in a single operation, it will lead to a shortening of the test time and labor saving in the tests, and the user benefits will be great. For example, in the case of identification tests, consider using mass spectrometry by MALDI-TOF, which is becoming more widespread. In MALDI, a dried sample is irradiated with a laser to perform desorption ionization of the sample. Therefore, it is preferable that the amount of moisture in the sample is small, and from the perspective of detection sensitivity, 5 a quantity of bacteria of 10

[0005] CFU (Colony forming unit) or more is required. 5 ~10 6A bacterial solution adjusted in concentration within the range of CFU / mL is used. When preparing this bacterial solution, it is common to use a method called the McFarland turbidity method. To accurately adjust the bacterial concentration, it is necessary to prepare several hundred μL of a solution with a concentration of 10 8 CFU / mL. Therefore, the amount of bacteria required is larger compared to the case of identification tests. Thus, in order to rapidly conduct a series of bacterial tests, it is necessary to extract bacteria from a blood culture positive sample with a relatively large volume of several mL or more, and obtain a plurality of samples with different solid and liquid forms as well as different amounts of bacteria all at once.

[0006] Patent Document 1 discloses a technique for separating bacteria from a blood sample and extracting the proteins of the bacteria. Patent Document 2 also discloses a technique for performing degradation of blood cells by protease and swelling treatment with a hypotonic solution, and selectively destroying only the blood cell components using a surfactant. Patent Document 3 further discloses a technique for separating specific components from a blood sample.

Prior Art Documents

Patent Documents

[0007]

Patent Document 1

Patent Document 2

Patent Document 3

Summary of the Invention

Problems to be Solved by the Invention

[0008] According to the method of Patent Document 1, it is possible to obtain a sample suitable for identification by mass spectrometry using MALDI. However, since the treatment is performed with a reagent that destroys the cell wall of the bacteria, the sample prepared by this method does not contain live bacteria and cannot be used for susceptibility tests.

[0009] Furthermore, the method of Patent Document 2 makes it possible to sufficiently destroy and remove blood cell components. This process involves repeated centrifugation and numerous washing steps involving solution replacement, making it cumbersome and time-consuming. While obtaining a liquid sample is easy, obtaining a solid sample requires additional centrifugation and supernatant removal, increasing the number of operational steps. Furthermore, obtaining samples with different bacterial counts requires the user to control the amount of bacterial solution dispensed, and the accuracy of this process depends on the technique used, such as dispensing, and is also a cumbersome process.

[0010] Furthermore, the method of Patent Document 3 makes it possible to easily separate specific components from a blood sample in one step by rotating it like a top. However, it is difficult to obtain samples in different solid and liquid forms for use in a series of bacterial tests. Furthermore, because a filter made of a porous material is used to separate the blood sample, clogging is a problem, making it unsuitable for rapid processing of samples of several mL to 10 mL.

[0011] The methods disclosed in Patent Documents 1 and 2 disclose methods for extracting bacteria from blood samples, but neither method can simultaneously obtain a solid bacterial sample required for identification testing or a liquid high-concentration bacterial sample required for susceptibility testing. Furthermore, the method disclosed in Patent Document 3 allows for a simple pretreatment of blood samples, but poses a challenge in simultaneously obtaining samples in different solid and liquid forms from a large volume of sample.

[0012] The present invention has been made in view of the above circumstances, and provides a collection method, an inspection method, a container, a centrifuge, and an inspection system that allow samples of different morphologies to be obtained from a specimen at once in a simple and stable manner. [Means for solving the problem]

[0013] In order to solve the above problems, the collection method of the present invention includes introducing a sample into a first storage section of a container including a first storage section having a first flow path and a second flow path of different heights on a side surface thereof, a second storage section connected to the first storage section via the first flow path, and a third storage section connected to the first storage section via the second flow path; centrifuging the sample introduced into the first storage section with a first centrifugal force to separate the sample into a first solution containing a first component and a second solution containing a second component; The method includes centrifuging the sample stored at a second centrifugal force greater than the first centrifugal force, and transferring the first solution to a second storage unit via a first flow path; centrifuging the second solution stored in the first storage unit at a third centrifugal force, and separating the second solution into a third solution containing the first component and a fourth solution containing the second component; and centrifuging the second solution stored in the first storage unit at a fourth centrifugal force greater than the third centrifugal force, and transferring the third solution to a third storage unit via the second flow path. [Effects of the Invention]

[0014] According to the present invention, it is possible to obtain samples with different morphologies from a sample at once by a simple and stable method.

[0015] Further features related to the present invention will become apparent from the description of this specification and the accompanying drawings. Furthermore, problems, configurations, and effects other than those described above will become apparent from the following description of the embodiments. The description of the present invention is merely a typical example and does not limit the scope of the claims or application examples in any sense. [Brief explanation of the drawings]

[0016] [Figure 1] FIG. 2 is a cross-sectional view of the container of Example 1. [Figure 2] 1 is a diagram showing the transition of a blood sample in a container, illustrating a method for obtaining a sample of a different form from a blood sample introduced into the container in Example 1. FIG. [Figure 3] FIG. 10 is a diagram showing the relationship between centrifugal acceleration and time in a method for obtaining samples of different forms from a blood sample. [Figure 4]It is a cross-sectional view showing the height of the flow path formed in the container of Example 1. [Figure 5] It is a block diagram showing the configuration of the inspection system of Example 1. [Figure 6] It is a block diagram showing the configuration of the centrifuge of Example 1. [Figure 7] It is a flowchart showing a method for obtaining samples in different forms from a blood sample and an inspection using the samples. [Figure 8] It is a block diagram showing the configuration of the inspection system of Example 2.

Mode for Carrying Out the Invention

[0017] Hereinafter, embodiments of the present invention will be described with reference to the drawings.

[0018] The accompanying drawings show specific embodiments based on the principles of the present invention, but these are for the purpose of understanding the present invention and are not used for limiting the interpretation of the present invention. In the embodiments and all the drawings for explaining the embodiments, those having the same function are denoted by the same reference numerals, and the repeated description thereof is omitted.

[0019] In the following examples, a collection method for obtaining samples in different forms at once from a blood sample containing microorganisms including impurities such as blood cells, and an inspection method for inspecting the first component from the samples obtained by this collection method will be described. Further, in the following examples, a container used in the collection method, a centrifuge including the container, and an inspection system including the centrifuge will be described.

[0020] In the following examples, by continuously separating blood cells and microorganisms in a blood sample, destroying the blood cells, and collecting and extracting the microorganisms, samples in solid and liquid forms and with different amounts of bacteria are obtained at once.

[0021] The "microorganisms" to be captured refer to various types of microorganisms including bacteria, actinomycetes, fungi, etc. However, viruses are not included in microorganisms. Specifically, they are the microorganisms that are the detection targets by the sterility test method in the pharmacopoeia, and the microorganisms such as pathogenic bacteria and pathogenic fungi that are the detection targets in hospital laboratories, etc. For example, the genus Escherichia (specific example: Escherichia coli), the genus Staphylococcus (specific examples: Staphylococcus aureus, Staphylococcus epidermidis), the genus Propionibacterium (specific example: Propionibacterium acnes), the genus Micrococcus, the genus Streptococcus (specific examples: Streptococcus pyogenes, Streptococcus pneumoniae), the genus Enterococcus (specific examples: Enterococcus faecium, Enterococcus faecalis), the genus Neisseria (specific examples: Neisseria gonorrhoeae, Neisseria meningitidis), the genus Moraxella, the genus Shigella (specific example: Shigella dysenteriae), the genus Salmonella (specific examples: Salmonella typhi, Salmonella paratyphi A, Salmonella enteritidis), the genus Citrobacter, the genus Klebsiella (specific example: Klebsiella pneumoniae), the genus Enterobacter, the genus Serratia (specific example: Serratia marcescens), the genus Proteus, the genus Providencia, the genus Morganella, the genus Yersinia (specific example: Yersinia pestis), the genus Vibrio (specific examples: Vibrio cholerae, Vibrio parahaemolyticus, Vibrio vulnificus, Vibrio mimicus), the genus Aeromonas, the genus Pseudomonas (specific example: Pseudomonas aeruginosa), the genus Acinetobacter (specific example: Acinetobacter baumannii), the genus Alcaligenes, the genus Agrobacterium, the genus Flavobacterium,Bacteria and fungi such as Haemophilus (specific example: Haemophilus influenzae), Pasteurella, Francisella, Bordetella (specific example: Bordetella pertussis), Eikenella, Brucella, Streptobacillus, Actinobacillus, Legionella (specific example: Legionella pneumophila), Bacillus (specific examples: Bacillus subtilis, Bacillus anthracis, Bacillus cereus), Corynebacterium (specific example: Corynebacterium diphtheriae), Lactobacillus, Listeria, Erysipelothrix, Nocardia, Actinomyces, Clostridium (specific examples: Clostridium perfringens, Clostridium sporogenes), Bacteroides (specific example: Bacteroides fragilis), Fusobacterium, Mycobacterium (specific example: Mycobacterium tuberculosis), Campylobacter, Helicobacter (specific example: Helicobacter pylori), Spirillum, Treponema, Borrelia, Leptospira, Mycoplasma (specific example: Mycoplasma pneumoniae), Aspergillus (specific examples: Aspergillus niger, Aspergillus brasiliensis), yeast (specific example: Candida albicans) are included, but other microorganisms can also be the collection targets.,

[0022] The blood sample is not particularly limited as long as it is a sample containing blood cells. In particular, it includes biological samples derived from a living body and samples suspected of being contaminated by microorganisms. For example, it can be various samples such as blood, urine, bone marrow fluid, breast milk, amniotic fluid, biopsy tissue, cell culture solution, and cell culture supernatant. Also, the origin of the blood sample is not particularly limited and can be derived from any biological species. For example, a blood sample derived from at least one of various types of organisms such as animals, plants, and insects is used as the test sample. When the blood sample is a liquid sample, it can be used as it is, or diluted or concentrated with a solvent. When the blood sample is a solid sample, it may be suspended in a solvent, homogenized with a pulverizer or the like, or the supernatant obtained by stirring with a solvent may be used. The blood sample may be diluted with an appropriate medium or physiological saline, or may be subjected to pretreatment or the like.

[0023] "Collection" means separating microorganisms from a solution containing blood cells, concentrating the microorganisms contained in the solution, etc. The concentration of microorganisms that may be contained in the sample is not particularly limited.

[0024] <Container> Referring to FIG. 1, the configuration of the container 1 of Example 1 will be described. The container 1 includes a first accommodating portion 100 into which a blood sample can be introduced, a second accommodating portion 101 for collecting solid bacteria, and a third accommodating portion 102 for collecting liquid bacteria.

[0025] A blood sample containing blood components is introduced into the first accommodating portion 100. Specifically, a sample that has become positive in a blood culture test is introduced. Therefore, the constituent components of the introduced sample may include blood components such as red blood cells, white blood cells, and platelets, a medium for bacterial growth, and bacteria.

[0026] A first flow path 103 and a second flow path 104 are provided on the side of the first storage unit 100. The first storage unit 100 is connected to the second storage unit 101 via the first flow path 103. The first storage unit 100 is also connected to the third storage unit 102 via the second flow path 104. The first flow path 103 is located higher than the second flow path 104. This allows a blood sample located higher than the first flow path 103 to move to the second storage unit 101 via the first flow path 103. Also, a blood sample located higher than the second flow path 104 to move to the third storage unit 102 via the second flow path 104.

[0027] The second storage unit 101 is provided with a filtration filter 105. The second storage unit 101 has a portion where the filtration filter 105 is installed. The filtration filter 105 is a filtering member that separates the solution that has moved from the first storage unit 100 to the second storage unit 101 into microorganisms and liquid components. The second storage unit 101 also has a waste liquid reservoir 106 formed therein. The filtration filter 105 captures only solid bacterial components, and the liquid components of the solution stored in the second storage unit 101 move to the waste liquid reservoir 106.

[0028] To prevent contamination, the receptacles 100, 101, and 102 are preferably sealed. In the container 1 of Example 1, for example, the first receptacle 100 is covered with a lid 107. Because a blood sample obtained from a blood culture bottle is introduced into the first receptacle 100, the lid 107 is preferably a lid such as a rubber stopper or a seal stopper. Furthermore, the second receptacle 101 and the third receptacle 102 are covered with lids 108. The second receptacle 101 and the third receptacle 102 accumulate the collected bacteria. Because the collected bacteria are recovered using a needle or rod with a sharp tip and a pipette tip or syringe, the lid is preferably made of a material that can be easily peeled off, such as a seal-type lid.

[0029] <Microorganism collection method> With reference to FIG. 2, a method for capturing microorganisms (bacteria in Example 1) from a blood sample will be described.

[0030] As shown in FIG. 2(a), a blood sample 200 is introduced into the first receptacle 100 of the container 1. The blood sample 200 may be introduced into the first receptacle 100 by a user or a robot. The blood sample 200 is preferably introduced to a position sufficiently higher than the position of the first flow channel 103. Next, a reagent 201 for destroying red blood cells (e.g., a blood cell destruction reagent) is introduced into the third receptacle 102. The reagent 201 may be introduced into the third receptacle 102 by a user or a robot. The reagent 201 is, for example, a surfactant. The surfactant preferably has a composition and concentration that does not affect bacterial cell membranes but destroys red blood cell membranes. The surfactant may be introduced after the blood sample 200 is introduced, or may be introduced into the third receptacle 102 before the blood sample 200 is introduced.

[0031] The container 1 containing the blood sample 200 is placed in a centrifugal device. The container 1 is then centrifuged at various centrifugal accelerations in the order shown in Figures 2(b) to 2(e). The centrifuged container 1 is gradually tilted and is ultimately positioned so that centrifugal force is applied toward the bottom of the container 1. At this time, the directions of the first flow path 103 and the second flow path 104 of the container 1 are different from the direction in which the centrifugal force is applied. The centrifugal acceleration applied to the container 1 and the centrifugation time will be shown in detail in Figure 3, which will be described later.

[0032] First, the blood sample 200 contained in the container 1 is centrifuged at low acceleration. As a result, as shown in Fig. 2(b), the blood sample 200 is separated into a bacterial solution (first solution) 202 containing bacteria (first component) and a solution (second solution) 203 containing heavy particles such as red blood cells and white blood cells (second component). In addition to bacteria, the bacterial solution 202 contains plasma, culture medium, etc., and in some cases may contain red blood cells and white blood cells that have not been completely separated.

[0033] Next, the blood sample 200 is centrifuged at an acceleration slightly higher than the initial low-acceleration centrifugation. As a result, as shown in FIG. 2(c), the centrifugal force exceeds the liquid pressure in the first flow path 103, and the bacteria liquid 202 moves to the second container 101 via the first flow path 103. The bacteria liquid 202 then contacts the filter 105, and the solid components, bacteria (first sample) 204, are captured on the filter 105. The filter 105 may be of any type, but is preferably a membrane filter with a diameter of 0.1 μm or less, and more preferably a hydrophilic filter made of a material such as PVDF. The filter 105 may have a diameter of several μm or less, as long as it can capture the bacteria 204. The liquid component 205 that passes through the filter 105, including plasma, culture medium, and proteins that are significantly smaller than bacteria, moves to the waste liquid reservoir 106. In this way, bacteria 204 contained in bacteria liquid 202 stored in second storage section 101 are captured on filter 105.

[0034] Next, the solution 203 is centrifuged at the same low acceleration as the initial step, and as a result, as shown in Fig. 2(d), the solution 203 in the first container 100 is separated into a bacterial solution (third solution) 206 containing bacteria and a blood cell component (fourth solution) 207 containing heavy particles such as red blood cells and white blood cells.

[0035] Finally, the solution 203 is centrifuged at the highest acceleration. Then, as shown in FIG. 2(e), the centrifugal force exceeds the liquid pressure in the second flow path 104, and the bacteria liquid 206 moves to the third storage section 102 via the second flow path 104. The bacteria liquid 206 that has moved to the third storage section 102 is mixed with the reagent 201, and the trace amount of blood cell components that have not been separated by the centrifugation and remain are destroyed. Thereafter, by subsequent centrifugation, the bacteria liquid 206 that has moved to the third storage section 102 is separated into a liquid component 208 and bacteria (second sample) 209. The bacteria 209 accumulate on the bottom surface of the third storage section 102 and can be collected.

[0036] The following method can be considered as a method for improving the accuracy of the analysis. For example, the bacteria 204 collected in the second container 101 and the bacteria 209 collected in the third container 102 may be contaminated with impurities contained in the blood sample 200. In this case, for example, the liquid component 208 is first removed from the state shown in FIG. 2(e). Then, an appropriate amount of surfactant or physiological saline is dropped onto the bacteria 204 and the bacteria 209, and they are centrifuged again at high acceleration to wash them, thereby making it possible to obtain a bacterial sample with a higher purity without complicated operations.

[0037] Next, the relationship between the centrifugal acceleration applied to the container 1 and the centrifugal time will be described with reference to Fig. 3. The states of the container 1 at times (a) to (e) in Fig. 3 correspond to the states (a) to (e) in Fig. 2.

[0038] First, the blood sample 200 is centrifuged at a centrifugal acceleration (first centrifugal force) x1 for a period of time t1. At this time, the centrifugal acceleration x1 must be set so that the pressure in the first flow path 103 is greater than the centrifugal force, so that the blood sample 200 in the first container 100 does not move to the second container 101 via the first flow path 103. Furthermore, since the centrifugation must be performed until the interface position between the bacteria liquid 202 and the solution 203 is lower than the height of the first flow path 103, the centrifugal acceleration time t1 is determined based on the sedimentation rate of the blood cells and the height of the first flow path 103. Specifically, the centrifugal acceleration x1 is preferably 20 to 100 G, for example, although it is affected by the diameter of the first flow path 103, the material of the container 1, and the physical properties (e.g., viscosity) of the blood sample 200.

[0039] Next, the blood sample 200 is centrifuged at a centrifugal acceleration (second centrifugal force) x2 for a time t2. The centrifugal acceleration x2 needs to be greater than the centrifugal acceleration x1. The centrifugal acceleration x2 is set such that the centrifugal force is greater than the pressure in the first flow path 103. Then, in the first flow path 103, it becomes impossible to hold the liquid, and the bacterial solution 202 moves to the second storage portion 101. Note that the pressure in the first flow path 103 is determined by surface tension in addition to the diameter and length of the first flow path 103. In the state of Fig. 2(b), the liquids in contact with the first flow path 103 and the second flow path 104 are different. The first flow path 103 is in contact with the bacterial solution 202 that contains almost no blood cells, and the second flow path 104 is in contact with the solution 203 that contains blood cells. Thus, since the surface tensions of the liquid in contact with the first flow path 103 and the liquid in contact with the second flow path 104 are different, the solution 203 does not move from the second flow path 104 that is in contact with the liquid having a greater surface tension.

[0040] Next, the solution 203 is centrifuged at a centrifugal acceleration (third centrifugal force) x1 for a time t3. At this time, it is necessary to set the centrifugal acceleration x1 such that the pressure in the second flow path 104 is greater than the centrifugal force so that the liquid does not move from the first storage portion 100 through the second flow path 104 to the third storage portion 102. Also, since it is necessary to perform centrifugation until the interface position between the bacterial solution 206 and the blood cell component 207 becomes lower than the height of the second flow path 104, the centrifugation time t3 is determined based on the sedimentation rate of the blood cells and the height of the second flow path 104.

[0041] Finally, the solution 203 is centrifuged at a centrifugal acceleration (fourth centrifugal force) x3 for a time t4. In order to quickly move the bacterial solution 206 from the second flow path 104 to the third storage portion 102, the centrifugal acceleration x3 is preferably a sufficiently high value. However, considering that it may affect the growth of bacteria in the bacterial solution 206, 10 4It is preferable to set the centrifugal acceleration as high as possible within a range not exceeding G. When centrifugation is performed at a centrifugal acceleration of x3, bacteria 204 and bacteria 209 are collected in second storage section 101 and third storage section 102, respectively. Solid bacteria 204 are collected on filtration filter 105 provided in second storage section 101, and a high concentration of bacteria 209 is collected on the bottom surface of third storage section 102.

[0042] <Height of the flow path formed in the container 1> The heights of the first channel 103 and the second channel 104 formed in the container 1 will be described with reference to FIG. 4. The heights of the first channel 103 and the second channel 104 are different. The reason for this is to automatically collect different forms of bacteria used in different tests. Two tests are considered: an identification test using MALDI and a susceptibility test. For example, in the case of an identification test using MALDI, 5 ~10 6 The susceptibility test requires 10 CFU or more of bacteria. 5 ~10 6 A bacterial solution with a CFU / mL concentration is required. For example, a concentration of 1.5 x 10 8 A few hundred μL of bacterial solution adjusted to a concentration of CFU / mL (equivalent to 0.5 McFarland) is prepared, and then diluted 100 to several hundred times before use in the test.

[0043] To consider the heights of the first flow channel 103 and the second flow channel 104, a positive blood culture sample will be considered. The volume of the positive blood culture sample introduced into the first container 100 was set to 10 mL. The concentration of bacteria contained in the positive blood culture sample varies depending on the time until the blood culture became positive, the time elapsed since the positive blood culture, the type of bacteria, and the strain. 7 ~10 9 Therefore, the bacterial concentration that is expected to be the most difficult to pretreat is in the range of 10 CFU / mL. 7 The case of CFU / mL was considered.

[0044] As a premise, the heights of the first flow path 103 and the second flow path 104 are as shown in FIG. 4. y1 is the height from the bottom surface of the first storage section 100 to the second flow path 104. y2 is the height from the second flow path 104 to the first flow path 103. y3 is the height from the first flow path 103 to the scale line 400 indicating the specified value. y1 + y2 + y3 is the height of the blood sample 200 when the entire volume of the expected blood sample 200, i.e., 10 mL, is placed. The above-mentioned visible scale line 400 is provided on the container 1 at a height position of y1 + y2 + y3. It is necessary to introduce at least the specified value or more of the blood sample 200 into the first storage section 100.

[0045] First, the height y1 from the bottom of the first container 100 to the second flow path 104 is determined as follows. Because centrifuged red blood cells accumulate in the first container 100, the volume of the first container 100 from the bottom to the height y1 must be equal to or greater than the volume of red blood cells in the blood sample 200. Here, the hematocrit value, which indicates the percentage of red blood cells in blood, ranges from 30 to 55%, depending on gender and health status. Furthermore, the blood sample 200 introduced into the first container 100 contains, in addition to the patient's blood, approximately two-thirds of its total volume is made up of culture medium components introduced beforehand. Therefore, the percentage of red blood cells in a positive blood culture sample can be said to be approximately one-sixth at most. Therefore, for more efficient blood sample pretreatment, the height y1 is preferably 16.7% or more of the specified height of the blood sample 200 introduced, e.g., 20%.

[0046] Next, the height of y3 is determined as follows: 7 In the case of CFU / mL, if there is 1 mL of sample, the number is 10 7 This allows extraction of CFU of bacteria. This is more than 10 times the amount of bacteria required for MALDI, and it is believed that a sufficient amount of bacteria can be extracted. Therefore, the height of y3 is preferably about 10% of the specified height of the input volume of blood sample 200.

[0047] From the above calculation, the height of y2 is automatically determined. In other words, the height of y2 is about 70% of the specified height of the blood sample 200 introduced. In this case, the amount of bacteria extracted near the bottom surface of the third storage section 102 is 10 7 CFU / mL x 7mL = 7 x 10 7 In actual operation, such a small amount of highly concentrated bacterial solution is diluted to several hundred μL, for example, 200 μL, to adjust the concentration, and the result is 1.5 × 10 CFU. 8 CFU / mL (0.5 McFarland equivalent). This calculation shows 1.5 x 10 8 For a bacterial solution requiring CFU / mL, 3.5 x 10 8 Since bacterial solution can be extracted at CFU / mL, it is possible to extract a sufficient amount even taking into account the loss of bacteria during isolation.

[0048] An example of this calculation is the assumption that the bacterial concentration in a positive blood culture sample is low; in reality, the bacterial concentration in a positive blood culture sample is 10 8 The number is often higher than CFU / mL, allowing bacteria to be extracted with ease.

[0049] In addition to the heights of the first flow channel 103 and the second flow channel 104, the elements that characterize the container 1 include the following.

[0050] Container 1 can use a container into which a blood sample 200 can be introduced and subjected to centrifugation. For example, the volume of the first accommodating portion 100 can be 1 to 20 mL, preferably 8 to 15 mL. The sum of the volume of the second accommodating portion 101 and the volume of the third accommodating portion 102 only needs to be equal to or greater than the volume of the first accommodating portion 100. Also, since the amount of bacteria collected in the third accommodating portion 102 is larger than that in the second accommodating portion 101, it is preferable that the volume of the third accommodating portion 102 is larger. Further, the material of the container 1 is not particularly limited as long as it is a material suitable for operations such as centrifugation. For example, the container 1 is preferably made of a hydrophobic material. This is because when the affinity between the container 1 and the blood sample 200 is good, the blood sample 200 may be moved from the first flow path 103 or the second flow path 104 before centrifugation due to surface tension. For example, the container 1 is preferably made of a material such as an acrylic resin, an ABS resin, polypropylene, polystyrene, or polyethylene, and is made using a 3D printer or injection molding. Alternatively, the container 1 may be made by cutting using aluminum or stainless steel. Chemical treatment may be performed on the container 1 to make it hydrophobic and surface modification may be carried out. Note that the container 1 may be opaque, but when it is transparent, it is easier to visually recognize the internal state or perform optical measurement or imaging by a device. For easy detection of excess or deficiency of the liquid volume, clogging of the sample, and detection of foreign substances in the sample, the container 1 is preferably transparent. From the perspective of contamination, the container 1 is preferably disposable, but for example, there is no problem in repeatedly using it after washing and sterilization treatment.

[0051] The outer shape of the container 1 can be any shape as long as it can be centrifuged. The bottom surface portion where the filtration filter 105 of the second accommodating portion 101 is installed and the bottom surface portion that accommodates the reagent of the third accommodating portion 102 are preferably in a mortar shape in order to efficiently collect bacteria 204 and 209.

[0052] The first flow path 103 and the second flow path 104 can have any shape. The first flow path 103 and the second flow path 104 are, for example, columnar with a circular cross-sectional shape or rectangular parallelepiped-shaped with a rectangular cross-sectional shape. The diameters of the first flow path 103 and the second flow path 104 can be considered as follows. Blood contains red blood cells, white blood cells, and platelets, and the largest among them is the white blood cell with a diameter of 6 - 30 μm. If the diameters of the first flow path 103 and the second flow path 104 are smaller than this, instead of the effect of the capillary valve, blood cells may clog the first flow path 103 and the second flow path 104, and the liquid may not be released. Therefore, although it also depends on the surface affinity between the blood sample 200 and the container 1, in terms of continuously separating blood cells from the liquid component and moving them from the first flow path 103 and the second flow path 104, the range of the optimal value of the diameter of the realistic first flow path 103 and the second flow path 104 is 10 - 100 μm. Also, the diameter of the first flow path 103 and the diameter of the second flow path 104 may be different, but preferably they have the same diameter.

[0053] The reagent 201 introduced into the third accommodating portion 102 is not particularly limited as long as it can destroy blood cells without affecting the growth of microorganisms. For example, the reagent 201 preferably contains at least one surfactant. Examples of the surfactant include, but are not limited to, an anionic surfactant having a hydrophilic and a hydrophobic part and the hydrophobic part being a chain hydrocarbon, or a surfactant having a hydrophilic and a hydrophobic part and the hydrophobic part having a cyclic hydrocarbon, or a combination of both. Specifically, examples of the former include sodium dodecyl sulfate, lithium dodecyl sulfate, and sodium N-lauroyl sarcosinate, and examples of the latter include saponin, sodium cholate, sodium deoxycholate, 3-[(3-cholamidopropyl)dimethylammonio]-1-propanesulfonate, and 3-[(3-cholamidopropyl)dimethylammonio]-2-hydroxy-1-propanesulfonate.

[0054] <Inspection System> Referring to FIG. 5, an inspection system 500 for automatically performing a bacterial inspection will be described. The inspection system 500 includes a centrifuge 501, a sampling device 502, an application device 503, a concentration measurement device 504, an identification inspection device 505, a susceptibility inspection device 506, and a control PC 508. Each device is connected by a signal line 507, and the control PC 508 controls the operation of each device.

[0055] The centrifuge 501 can accommodate the above-described one or more containers 1, centrifuges the blood sample 200 introduced into the first accommodating portion 100 of the mounted container 1, and separates the bacteria 204 used in the identification inspection device 505 and the bacteria 209 used in the susceptibility inspection device 506. The centrifuge 501 centrifuges the blood sample 200 at at least three accelerations set for centrifugation of the blood sample 200, movement of the bacterial liquid 202 to the second accommodating portion 101, and movement of the bacterial liquid 206 to the third accommodating portion 102. The container 1 is transported to the sampling device 502 by a transport mechanism 509. The user may transport the container 1 from the centrifuge 501 to the sampling device 502.

[0056] The sampling device 502 acquires the bacteria 204 collected in the second accommodating portion 101 of the container 1 using, for example, a sharp specimen sampling rod 510 at the tip, and acquires the bacteria 209 collected in the third accommodating portion 102 of the container 1 using a syringe 511. The acquired bacteria 204 and bacteria 209 are sent to different devices respectively.

[0057] In the application device 503, solid bacteria are applied to the MALDI target plate used in mass spectrometry for identification inspection, and a matrix reagent is applied.

[0058] In the concentration measurement device 504, for example, the concentration of the bacterial liquid is measured by measuring the McFarland turbidity.

[0059] The pretreatment device 512 is a device that combines the above-mentioned centrifuge device 501, fraction collection device 502, coating device 503, and concentration measurement device 504. However, the coating device 503 does not necessarily have to be present as a necessary component. For example, the fraction collection device 502 may return the specimen collection rod 510 from which the bacteria 204 has been collected to the user, and the user may apply the bacteria 204 to the target plate by themselves.

[0060] The concentration value measured by the pretreatment device 512 may be displayed on a display attached to the control PC 508 to inform the user. If the concentration value is equal to or higher than the target value, the user may be notified, for example, and the user may adjust the concentration to the target value, thereby maintaining the reliability of the test. If the concentration is equal to or lower than the target value, an error flag may be displayed, and the pretreatment may be performed again, or the bacterial liquid may be cultured again before being used for testing.

[0061] As an additional function, the fraction collection device 502 may acquire external appearance information of the container 1, for example, by using concentration measurement or image measurement, to confirm whether the preceding process is being performed normally. For example, if the preceding process is being performed normally, the liquid level of the blood sample 200 in the first receptacle 100 will be approximately the same as the level of the second channel 104, as shown in FIG. 2(e). However, if the preceding process is stopped midway, the liquid level of the blood sample 200 in the first receptacle 100 will be sufficiently higher than the level of the second channel 104. Based on this information, the fraction collection device 502 can verify whether the preceding process has been completed normally.

[0062] The sample processed by the pretreatment device 512 is once returned to the user, and after additional processing steps or the like are performed as necessary, it is inspected by the identification inspection device 505 and the susceptibility inspection device 506. The identification inspection device 505 determines the type of bacteria using the sample obtained from the bacteria 204. In addition, the susceptibility inspection device 506 determines the degree of bacterial growth using the sample obtained from the bacteria 209. In the case of identification inspection, in some cases, after the user applies bacteria and matrix reagent to the target plate by themselves, the device identifies the bacterial species. In the case of susceptibility inspection, for example, the bacterial solution is adjusted to the target concentration and dispensed into a 96-well plate or the like, and the susceptibility inspection device 506 determines the growth and MIC. The progress and determination results of the bacterial species identification and susceptibility inspection are displayed on the display attached to the control PC 508, and the results are notified to the user. In addition, the control PC 508 preferably inputs parameters of each device such as the acceleration and time of centrifugation in the centrifuge device 501 and the target value of the bacterial concentration in the concentration measurement device 504 according to the blood sample 200, and can change the processing content according to the values.

[0063] <Centrifuge device> The centrifugal device 501 will be described in detail with reference to FIG. 6. As shown in FIG. 6, the centrifugal device 501 includes a container mounting unit 601, a drive unit 602, and a control board 603. One or more containers 1 are mounted on the container mounting unit 601. The drive unit 602 is an actuator that applies centrifugal force to the blood sample 200 in the container 1 mounted on the container mounting unit 601. The control board 603 is a control unit that controls the operation of the drive unit 602. The control board 603 includes a processor 610, a main memory device 611, an auxiliary memory device 612, an input / output I / F 613, and a bus 614 that communicatively connects the various components of the control board 603. The input / output I / F 613 is communicatively connected to a control PC 508 via a signal line 507 and receives instructions from the control PC 508. The processor 610 generates a drive signal for driving the drive unit 602 in accordance with the instructions from the control PC 508. The input / output I / F 613 is communicatively connected to the driving unit 602, and outputs a driving signal generated by the processor 610 to the driving unit 602. The driving unit 602 centrifuges the container 1 at various centrifugal accelerations in accordance with the driving signal.

[0064] <Bacteria testing method> The bacteria inspection method of Example 1 will be described with reference to Fig. 7. Each process from S71 to S74 in Fig. 7 is executed by the centrifugal device 501 in response to a command from the control board 603, which is the computer system of the centrifugal device 501.

[0065] First, in step S70, a blood sample 200 is introduced into the first storage section 100 of the container 1. The process of introducing the blood sample 200 into the container 1 may be performed by a user or a robot. Note that a reagent 201 such as a surfactant that disrupts blood cells may be introduced into the third storage section 102 in step S70, or may be introduced into the third storage section 102 before step S70.

[0066] 7, steps S71 to S74 are shown individually, but the series of processes of steps S71 to S74 are automatically and continuously performed by centrifugal device 501. In step S71, container 1 containing blood sample 200 is centrifuged at low acceleration to separate blood sample 200 into bacteria liquid 202 and solution 203. In step S72, container 1 containing blood sample 200 is centrifuged at medium acceleration to move bacteria liquid 202 from first storage section 100 to second storage section 101 via first flow path 103. Then, liquid component 205 is removed from bacteria liquid 202 moved to second storage section 101 by filter 105, and bacteria 204 are collected. In step S73, container 1 containing solution 203 is centrifuged at low acceleration to separate solution 203 into bacteria liquid 206 and blood cell component 207. Then, in step S74, the container 1 containing the solution 203 is centrifuged at high acceleration to move the bacteria liquid 206 from the first container 100 to the third container 102 via the second flow path 104. In the third container 102, the red blood cells remaining in the bacteria liquid 206 are destroyed by the reagent 201 and eluted into the liquid. As a result, only the bacterial components accumulate on the bottom surface of the third container 102.

[0067] In step S75, the identification test device 505 performs an identification test using the bacteria 204 collected in the second storage unit 101. For example, for an identification test using MALDI, a mass of solid bacteria 204 collected on the filtration filter 105 of the second storage unit 101 is obtained, applied to a target plate, and then an appropriate matrix reagent is dropped onto the target plate, dried, and then an identification test is performed. To improve the detection sensitivity of the identification test, there is no problem if a protein extraction process using ethanol, formic acid, acetonitrile, or the like is performed, as in the case of a typical MALDI method.

[0068] In step S76, the susceptibility testing device 506 acquires the high-concentration bacteria liquid accumulated on the bottom surface of the third container 102 and performs a susceptibility test. Specifically, the high-concentration bacteria 209 accumulated on the bottom surface is acquired using a pipette tip or the like, or the high-concentration bacteria 209 accumulated on the bottom surface is acquired after removing the supernatant. Then, the concentration of the bacteria liquid is adjusted by, for example, McFarland turbidimetry, and the susceptibility test is performed using the bacterial liquid.

[0069] In step S77, the results of the bacterial test are reported. Specifically, the identification test performed in S75 reports information on the bacterial species and the judgment score. In addition, the susceptibility test performed in S76 reports information such as the drug name, the drug's MIC (Minimum Inhibitory Concentration), and susceptibility / resistance to the laboratory technician and doctor.

[0070] <Effects of Example 1> In Example 1, a container 1 including a first storage unit 100 having a first flow path 103 and a second flow path 104 at different heights on its side, a second storage unit 101 connected to the first storage unit 100 via the first flow path 103, and a third storage unit 102 connected to the first storage unit 100 via the second flow path 104 is centrifuged at various centrifugal accelerations. This makes it possible to separate blood cells and bacteria from a blood sample 200 introduced into the first storage unit 100. Then, by centrifuging the blood sample 200 stored in the first storage unit 100, a bacteria liquid 202 can be moved to the second storage unit 101 via the first flow path 103, and a bacteria liquid 206 can be moved to the third storage unit 102 via the second flow path 104. This makes it possible to capture solid bacteria 204 and liquid bacteria 209 with different morphologies in the second storage section 101 and the third storage section 102. In this way, in Example 1, it is possible to simultaneously obtain bacterial samples with different morphologies to be used in identification tests and susceptibility tests using a simple and stable method.

[0071] As described above, after introducing a blood sample, a single series of centrifugation steps at a predetermined centrifugal acceleration extracts different bacterial samples. Solid bacterial samples with a small liquid component are suitable for MALDI mass spectrometry, while liquid bacterial samples are suitable for susceptibility testing. The amount of bacteria extracted can be controlled by adjusting the height of the first channel 103 and the second channel 104, allowing users to automatically obtain a bacterial sample of a specified amount or more suitable for each test without complex operations.

[0072] Sample preparation equivalent to the isolation of bacteria from colonies after isolation and culture and the preparation of a bacterial suspension, which normally takes about a day and night, can now be achieved in a single pretreatment process of about several tens of minutes. Conventional methods require repeated centrifugation due to multiple isolation, disruption, and washing steps, and it is not possible to obtain different samples for identification tests or susceptibility tests with a single pretreatment process. However, the present invention enables rapid testing by performing a series of steps continuously.

[0073] Furthermore, in the first embodiment, the solid bacteria 204 can be collected by the filtration filter 105 of the second storage section 101. This makes it possible to obtain the solid bacteria required for the identification test.

[0074] Furthermore, in Example 1, the reagent 201 introduced into the third container 102 does not affect the cell membranes of bacteria in the bacterial solution 206, but can destroy the cell membranes of red blood cells. This makes it possible to destroy the trace amounts of blood cell components remaining in the bacterial solution 206 that were not separated by centrifugation.

[0075] Furthermore, in Example 1, the heights of the first flow path 103 and the second flow path 104 are ⅙ or more of the height from the bottom surface of the first storage section 100 to a specified value, and the difference (y2) between the heights of the first flow path 103 and the second flow path 104 is ½ or more of the height from the bottom surface of the first storage section 100 to a specified value. It is possible to obtain the amount of bacterial sample required for identification tests and susceptibility tests from blood sample 200, which contains the patient's blood and about ⅔ of the total volume of culture medium components.

[0076] Furthermore, in Example 1, by centrifuging the solution 203 at a centrifugal acceleration x3 that is sufficiently greater than the centrifugal acceleration x2, it becomes possible to quickly move the bacteria liquid 206 from the second flow path 104 to the third storage section 102.

[0077] Furthermore, in the first embodiment, the type of bacteria can be determined using the bacteria 204, and the growth rate of the bacteria can be determined using the bacteria 209.

[0078] Furthermore, in Example 1, the bottom surfaces of the second storage section 101 and the third storage section 102 of the container 1 are each shaped like a mortar, so that the captured bacteria 204 and 209 can be efficiently obtained with a sharp needle or stick, a pipette tip, or a syringe.

[0079] Example 2 In Example 1, an example was described in which a user manually obtains the bacteria 204 collected in the second storage unit 101 and the bacteria 209 collected in the third storage unit 102 using a needle or a pipette tip. In Example 2, the bacteria 204 collected in the second storage unit 101 are automatically collected and applied, and the bacteria 209 collected in the third storage unit 102 are automatically collected, diluted, and the concentration is measured.

[0080] Similar to the first embodiment, the testing system 800 of the second embodiment includes a centrifugal device 501, a fraction collection device 502, a coating device 503, an identification testing device 505, a susceptibility testing device 506, and a control PC 508. The testing system 800 of the second embodiment also includes a concentration measurement and dilution device 801, a drive mechanism 802, a drive control mechanism 803, a gripping mechanism 804, and a transport device 809.

[0081] The drive mechanism 802 is controlled by a drive control mechanism 803. The drive mechanism 802 moves the gripping mechanism 804 from the centrifuge device 501 to the fraction collection device 502. The gripping mechanism 804 is configured to be able to grip a container 1, and grips the container 1 centrifuged by the centrifuge device 501. The drive mechanism 802 also moves the specimen collection rod 510 from the fraction collection device 502 to the application device 503. Furthermore, the drive mechanism 802 moves the syringe 511 from the fraction collection device 502 to the concentration measurement and dilution device 801. The sample prepared in the application device 503 is transported to the identification testing device 505 by the transport device 809. Furthermore, the sample whose concentration has been measured and diluted by the concentration measurement and dilution device 801 is transported to the susceptibility testing device 506 by the transport device 809.

[0082] <Effects of Example 2> The testing system 800 of the second embodiment can automatically perform the processes from centrifugation in the centrifugal device 501 to testing in the identification testing device 505 and testing in the susceptibility testing device 506 .

[0083] The present invention is not limited to the above-described embodiments and includes various modifications. The above-described embodiments have been described in detail to clearly explain the present invention, and the present invention is not necessarily limited to all of the configurations described. Furthermore, it is possible to replace part of the configuration of one embodiment with the configuration of another embodiment, or to add the configuration of another embodiment to the configuration of one embodiment. Furthermore, it is also possible to add, delete, or replace part of the configuration of each embodiment with other configurations. [Explanation of symbols]

[0084] 1...container, 100...first storage section, 101...second storage section, 102...third storage section, 103...first flow path, 104...second flow path, 105...filtration filter, 106...waste liquid reservoir, 107...lid, 108...lid, 200...blood sample, 201...reagent, 202...bacterial solution, 203...solution, 204...bacteria, 205...liquid component, 206...bacterial solution, 207...blood cell component, 208...liquid component, 209...bacteria, 400...scale mark, 500...testing system, 501...centrifuge, 502...fractionation device, 503...applicator, 504...concentration Measurement device, 505...identification testing device, 506...susceptibility testing device, 507...signal line, 508...control PC, 509...transport mechanism, 510...specimen collection rod, 511...syringe, 512...pretreatment device, 601...container mounting unit, 602...drive unit, 603...control board, 610...processor, 611...main memory device, 612...auxiliary memory device, 613...input / output I / F, 614...bus, 800...testing system, 801...concentration measurement and dilution device, 802...drive mechanism, 803...drive control mechanism, 804...gripping mechanism, 809...transport device

Claims

1. Introducing a sample into the first accommodating portion of a container comprising a first accommodating portion having a first channel and a second channel with different heights on a side surface, a second accommodating portion connected to the first accommodating portion via the first channel, and a third accommodating portion connected to the first accommodating portion via the second channel, Centrifuging the sample introduced into the first accommodating portion with a first centrifugal force to separate it into a first solution containing a first component and a second solution containing a second component, Centrifuging the sample accommodated in the first accommodating portion with a second centrifugal force greater than the first centrifugal force to move the first solution to the second accommodating portion via the first channel, Centrifuging the second solution accommodated in the first accommodating portion with a third centrifugal force to separate it into a third solution containing the first component and a fourth solution containing the second component, and Centrifuging the third solution and the fourth solution accommodated in the first accommodating portion with a fourth centrifugal force greater than the third centrifugal force to move the third solution to the third accommodating portion via the second channel, characterized in that the first channel is provided at a position higher than the second channel, introducing the sample into the first accommodating portion includes introducing the sample into the first accommodating portion to a prescribed value or more higher than the first channel, moving the first solution to the second accommodating portion includes moving an amount of the first solution determined based on the difference between the height of the sample introduced into the first accommodating portion and the height of the first channel to the second accommodating portion via the first channel, moving the third solution to the third accommodating portion includes moving an amount of the third solution determined based on the difference between the height of the first channel and the height of the second channel to the third accommodating portion via the second channel, the heights of the first channel and the second channel are 1 / 6 or more of the height from the bottom surface of the first accommodating portion to the prescribed value, the difference between the height of the first channel and the height of the second channel is 1 / 2 or more of the height from the bottom surface of the first accommodating portion to the prescribed value, A collection method characterized by the above.

2. Introducing a sample into the first accommodating portion of a container comprising a first accommodating portion having a first channel and a second channel with different heights on a side surface, a second accommodating portion connected to the first accommodating portion via the first channel, and a third accommodating portion connected to the first accommodating portion via the second channel, Centrifuging the sample introduced into the first accommodating portion with a first centrifugal force to separate it into a first solution containing a first component and a second solution containing a second component, Centrifuging the sample accommodated in the first accommodating portion with a second centrifugal force greater than the first centrifugal force to move the first solution to the second accommodating portion via the first channel, Centrifuging the second solution accommodated in the first accommodating portion with a third centrifugal force to separate it into a third solution containing the first component and a fourth solution containing the second component, and Centrifuging the third solution and the fourth solution accommodated in the first accommodating portion with a fourth centrifugal force greater than the third centrifugal force to move the third solution to the third accommodating portion via the second channel, having, the fourth centrifugal force being greater than the second centrifugal force, a collection method characterized by this.

3. In the collection method according to claim 1 or 2, separating the first solution into the first component and a liquid component by a filtering member provided in the second accommodating portion, further having this as a feature.

4. In the collection method according to claim 1 or 2, destroying the cell membrane of the second component contained in the third solution with a reagent accommodated in the third accommodating portion, further having this as a feature.

5. Obtaining a first sample from the first solution moved to the second accommodating portion by the collection method according to claim 1 or 2, Obtaining a second sample from the third solution moved to the third accommodating portion by the collection method according to claim 1 or 2, Determining the type of the first component using the first sample, and Determining the growth degree of the first component using the second sample, an inspection method characterized by having this.

6. A centrifuge device comprising a container, the container being capable of introducing a sample, and having a first accommodating portion having a first channel and a second channel with different heights on a side surface, a second accommodating portion connected to the first accommodating portion via the first channel, The first storage part and a third storage part connected via the second flow path, comprising: The centrifugal device further comprises: a drive part for centrifuging the sample introduced into the first storage part of the container; a control part for controlling the centrifugal force applied by the drive part to the sample, wherein the control part: controls the drive part to centrifuge the sample introduced into the first storage part with a first centrifugal force, and separates the sample into a first solution containing a first component and a second solution containing a second component; controls the drive part to centrifuge the sample contained in the first storage part with a second centrifugal force greater than the first centrifugal force, and moves the first solution to the second storage part via the first flow path; controls the drive part to centrifuge the second solution contained in the first storage part with a third centrifugal force, and separates it into a third solution containing the first component and a fourth solution containing the second component; controls the drive part to centrifuge the third solution and the fourth solution contained in the first storage part with a fourth centrifugal force greater than the third centrifugal force, and moves the third solution to the third storage part via the second flow path; wherein the control part: controls the drive part so that the fourth centrifugal force is greater than the second centrifugal force. A centrifugal device characterized by the above.

7. In the container, the second storage part has a part where a filtering member for separating the solution moved from the first storage part to the second storage part into a first component and a liquid component is installed. The centrifugal device according to claim 6, characterized by the above.

8. In the container, the third storage part has a part for storing a reagent that destroys the cell membrane of the second component contained in the solution moved from the first storage part to the third storage part. The centrifugal device according to claim 6, characterized by the above.

9. An inspection system comprising a centrifugal device equipped with a container, wherein the container: is capable of introducing a sample, and has a first storage part with a first flow path and a second flow path of different heights on the side surface; a second storage part connected to the first storage part via the first flow path; a third storage part connected to the first storage part via the second flow path; comprising: The centrifugal device further comprises: a drive part for centrifuging the sample introduced into the first storage part of the container; a control part for controlling the centrifugal force applied by the drive part to the sample, wherein the control part: Control the drive unit to centrifuge the sample introduced into the first storage unit at a first centrifugal force, and separate the sample into a first solution containing a first component and a second solution containing a second component. Control the drive unit to centrifuge the sample stored in the first storage unit at a second centrifugal force greater than the first centrifugal force, and move the first solution to the second storage unit through the first flow path. Control the drive unit to centrifuge the second solution stored in the first storage unit at a third centrifugal force, and separate it into a third solution containing the first component and a fourth solution containing the second component. Control the drive unit to centrifuge the third solution and the fourth solution stored in the first storage unit at a fourth centrifugal force greater than the third centrifugal force, and move the third solution to the third storage unit through the second flow path. The inspection system further includes A fractionation device that acquires a first sample from the second solution stored in the second storage unit of the container and acquires a second sample from the third solution stored in the third storage unit. An identification inspection device that determines the type of the first component using the first sample acquired by the fractionation device. A sensitivity inspection device that determines the growth degree of the first component using the second sample acquired by the fractionation device. An inspection system, characterized by comprising the above.

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