Nucleic acid analysis method and nucleic acid analysis device

Abstract

To provide a nucleic acid analysis method that is capable of reducing a reagent cost and improving through-put while performing accuracy assurance of nucleic acid analysis.SOLUTION: In a nucleic acid analysis method being an example of an embodiment, a first sample set including a test sample prepared from a first subject specimen and reagent, and at least one control sample prepared from at least one of a positive control and a negative control and reagent is fabricated, and a second sample set which includes a test sample prepared from a second subject specimen and reagent, and does not include at least one of the control samples included in the first sample set is fabricated. Nucleic acid amplification in the first sample set is measured by the first unit, nucleic acid amplification in the second sample set is measured by the second unit, and, a measurement result of each test sample included in the first and second sample sets is analyzed on the basis of a measurement result of the control sample included in at least the first sample set.SELECTED DRAWING: Figure 1

Description

[Technical Field]

[0001] The present invention relates to a nucleic acid analysis method and a nucleic acid analysis device. [Background technology]

[0002] The global COVID-19 pandemic has led to a sharp increase in demand for PCR tests for infectious viruses.

[0003] In a typical PCR measurement device, amplified nucleic acids are detected by repeatedly heating and cooling a plate with many wells that contains samples. Patent Document 1 discloses a method for amplifying nucleic acids of many samples in batches by storing multiple samples in a plate with multiple wells and repeatedly heating and cooling the plate. [Prior art documents] [Patent documents]

[0004] [Patent Document 1] Japanese Patent Application Laid-Open No. 2008-22732 Summary of the Invention [Problem to be solved by the invention]

[0005] As disclosed in Patent Document 1, in batch processing using well plates, a standard nucleic acid sample (also called a positive control) and a negative control are conventionally included in each batch to ensure the accuracy of nucleic acid analysis. If a batch always contains a fixed number of positive controls and negative controls, reagent costs for measuring the controls are incurable for each batch. Furthermore, the number of subject samples that can be measured in one batch is reduced by the number of controls included in one batch.

[0006] An object of the present invention is to provide a nucleic acid analysis method that enables reduction in reagent costs and improvement in throughput while ensuring the accuracy of nucleic acid analysis. [Means for solving the problem]

[0007] The nucleic acid analysis method of the present invention is characterized by preparing a first sample set including a test sample prepared from a first subject sample and a reagent, at least one of a positive control and a negative control, and a control sample prepared from the reagent; preparing a second sample set including a test sample prepared from a second subject sample and the reagent, but not including at least one of the control samples included in the first sample set; performing nucleic acid amplification on the first sample set and measuring the amplified nucleic acid; performing nucleic acid amplification on the second sample set and measuring the amplified nucleic acid; and analyzing the measurement results of each of the test samples included in the first and second sample sets based on the measurement results of at least the control sample included in the first sample set.

[0008] The nucleic acid analysis device of the present invention comprises a sample preparation device that prepares multiple sample sets, each including a test sample prepared from a subject's sample and a reagent; at least one unit that performs nucleic acid amplification on each of the multiple sample sets and measures the amplified nucleic acid; and a control unit.The sample preparation device, under the control of the control unit, prepares a first sample set that includes the test sample, at least one positive control and negative control, and a control sample prepared from the reagent, and a second sample set that includes the test sample but does not include at least one of the control samples included in the first sample set.The control unit analyzes the measurement results of each test sample included in the first and second sample sets based on the measurement results of at least the control sample included in the first sample set. [Effects of the Invention]

[0009] According to the present invention, it is possible to ensure the accuracy of nucleic acid analysis while reducing reagent costs and improving throughput. [Brief explanation of the drawings]

[0010] [Figure 1]1 is a schematic diagram of a nucleic acid analyzer according to an embodiment. [Figure 2] FIG. 1 is a perspective view of a PCR unit. [Figure 3] FIG. 1 is a perspective view of an eight-tube assembly. [Figure 4] 1 is a block diagram showing a configuration of a nucleic acid analyzer according to an embodiment; [Figure 5] 10 is a flowchart showing an example of the operation of the nucleic acid analyzer. [Figure 6] 10 is a flowchart showing an example of the operation of the nucleic acid analyzer. [Figure 7] 10 is a flowchart showing an example of the operation of the nucleic acid analyzer. [Figure 8] 10 is a flowchart showing an example of the operation of the nucleic acid analyzer. [Figure 9] 10 is a flowchart showing an example of a procedure for analyzing measurement results (measurement data). [Figure 10] FIG. 10 is a diagram illustrating an example of an analysis result. [Figure 11] FIG. 1 shows an example of sample arrangement in a PCR unit group. [Figure 12] 1 is a flowchart illustrating an example of a sample set creation process. [Figure 13] FIG. 10 is a diagram showing another example of sample arrangement in a PCR unit group. [Figure 14] 10 is a flowchart illustrating another example of a process for preparing a sample set. [Figure 15] FIG. 10 is a diagram showing another example of sample arrangement in a PCR unit group. [Figure 16] FIG. 10 is a diagram showing another example of sample arrangement in a PCR unit group. [Figure 17] FIG. 10 is a diagram showing another example of sample arrangement in a PCR unit group. [Figure 18] FIG. 10 shows a modified example of the arrangement of control samples in a PCR unit group. [Figure 19] FIG. 10 shows another modified example of the arrangement of control samples in a PCR unit group. [Figure 20]FIG. 10 shows another modified example of the arrangement of control samples in a PCR unit group. [Figure 21] FIG. 1 is a diagram showing a sample arrangement based on a conventional method. [Figure 22] FIG. 10 is a diagram showing a modified example of a nucleic acid analyzer. [Figure 23] FIG. 1 is a diagram illustrating an example of a detection device. DETAILED DESCRIPTION OF THE INVENTION

[0011] Hereinafter, an example of an embodiment of a nucleic acid analysis method and a nucleic acid analysis device according to the present invention will be described in detail with reference to the drawings. The embodiment described below is merely an example, and the present invention is not limited to the following embodiment. Furthermore, selective combinations of the components of the multiple embodiments and variations described below are within the scope of this disclosure.

[0012] As used herein, the term "sample" refers to a mixture prepared by mixing a specimen with a reagent. Furthermore, as used herein, the term "test sample" refers to a sample prepared using a subject specimen as the specimen, and is used to distinguish it from a "control sample" prepared using a positive control or negative control as the specimen. A "positive control sample" refers to a sample prepared using a positive control as the specimen. A "negative control sample" refers to a sample prepared using a negative control as the specimen. A "positive control sample" and a "negative control sample" are collectively referred to as "control samples."

[0013] As used herein, the term "sample set" refers to a unit consisting of a set of multiple samples, and one unit is defined as a number of samples that can be nucleic acid amplified by a single batch process using one unit or fewer.

[0014] As used herein, the term "QC group" refers to a collection of multiple samples prepared using a single common reagent. A "common reagent" may be a reagent obtained by a single preparation, such as a mixed reagent obtained by mixing a primer reagent and an enzyme reagent. In this case, the mixed reagent contains an amount corresponding to the number of tests, and multiple samples prepared using the mixed reagent form one QC group. A "common reagent" may also be a reagent contained in one vial. In this case, multiple samples prepared using the reagent contained in the common vial form one QC group. A "common reagent" may also be a reagent from the same manufacturing lot. In this case, multiple reagents contained in multiple different vials and assigned the same manufacturing lot form a common reagent, and multiple samples prepared using the reagent from the same manufacturing lot form one QC group.

[0015] Fig. 1 is a schematic diagram of a nucleic acid analyzer 1 according to an embodiment. As shown in Fig. 1, the nucleic acid analyzer 1 mainly comprises a sample preparation robot 11 that prepares a plurality of sample sets, each including a test sample prepared from a subject's specimen and a reagent; a PCR unit group 20 that measures the amplified nucleic acid of each sample in the plurality of sample sets; a measurement robot 22 that distributes the sample sets prepared by the sample preparation robot 11 to PCR units 21 in the PCR unit group 20; a robot controller 32 that controls each robot; a PCR controller 33 that controls the PCR units 21; and a system controller 31 that controls the overall operation of the nucleic acid analyzer 1. The PCR unit group 20 is composed of a plurality of PCR units 21.

[0016] The sample preparation robot 11 prepares a first sample set including a test sample and at least one control sample under the control of the robot control unit 32. The sample preparation robot 11 also prepares a second sample set including the test sample but excluding at least one of the control samples included in the first sample set. The control sample included in the first sample set is at least one of a positive control sample prepared from a positive control and a reagent, and a negative control sample prepared from a negative control and a reagent.

[0017] The PCR control unit 33 analyzes the measurement results of each test sample contained in the first and second sample sets based on the measurement results of the control sample contained in at least the first sample set. By utilizing the measurement results of the control sample contained in the first sample set not only for the analysis of the test samples in the first sample set but also for the analysis of the test samples in the second sample set, it is possible to reduce reagent costs and improve throughput.

[0018] The positive control sample is a sample containing a known concentration of nucleic acid to be tested (also referred to as target nucleic acid), and is a sample used to confirm the accuracy of nucleic acid amplification by the PCR unit 21. The positive control sample is generally called a standard nucleic acid sample, positive control, quality control sample, or QC specimen sample. The negative control sample is a sample that does not contain at least the nucleic acid to be tested, and is, for example, a sample used to confirm the presence or absence of contamination in each process of the nucleic acid analyzer 1. The negative control sample is generally called a negative control.

[0019] In one example, the nucleic acid analyzer 1 is used for virus testing based on a real-time PCR method. The principle of nucleic acid amplification is not limited to real-time PCR as long as it amplifies and analyzes nucleic acids, and may be, for example, an isothermal amplification method such as the Invader method, the LAMP method, the SmartAmp method, or the TMA method.

[0020] An example of a virus to be tested is SARS-CoV-2. The nucleic acid analyzer 1 may be installed, for example, at an airport and may constitute at least a part of a system that performs fully automated virus testing on samples collected from subjects who are scheduled to board an airplane.

[0021] In one example, the subject sample collected from the subject is saliva. The subject sample may be other types of samples derived from the respiratory system, such as throat swabs, nasal swabs, nasal discharge, sputum, or gargle. The subject sample may also be whole blood, serum, plasma, cerebrospinal fluid (CSF), pleural fluid, pericardial fluid, synovial fluid, urine, stool, or tissue sections.

[0022] In this embodiment, a nucleic acid extract obtained by extracting nucleic acids contained in saliva collected from a subject is contained in a well plate 3 as a subject sample and supplied to the nucleic acid analyzer 1 by a belt conveyor 2. One or more well plates 3 are sequentially supplied to the belt conveyor 2 in response to test requests. The well plate 3 has multiple wells, each of which contains an individual subject sample, which is a nucleic acid extract. A single well plate 3 contains multiple subject samples. The nucleic acid analyzer 1 collects the subject samples from the supplied well plates 3, prepares test samples from the subject samples and reagents, and performs nucleic acid amplification measurements of each test sample in parallel using multiple PCR units 21.

[0023] The sample preparation robot 11, under the control of the robot control unit 32, prepares test samples and control samples to prepare a sample set including multiple samples. The sample preparation robot 11 dispenses a predetermined amount of subject specimen from a well plate 3 supplied by a belt conveyor 2 and mixes the subject specimen with a reagent to prepare a test sample. The sample preparation robot 11 can also prepare positive and negative control samples. In this embodiment, each sample is prepared using an eight-tube array 40, which has eight connected containers 41, to prepare a sample set. In other words, the multiple samples contained in the eight-tube array 40 constitute one sample set.

[0024] The nucleic acid analyzer 1 includes a sample preparation robot 11, a belt conveyor 2, a reagent storage unit 12, a QC sample storage unit 13, a nozzle tip storage unit 14, and an 8-tube storage area 15, all of which are arranged around the robot.

[0025] The belt conveyor 2 is disposed in front of the sample preparation robot 11. The belt conveyor 2 transports well plates 3 containing multiple nucleic acid extracts as subject samples to a position accessible to the sample preparation robot 11. The well plates 3 have, for example, 96 wells. The belt conveyor 2 is equipped with a sensor 2a that detects newly inserted well plates 3. The sensor 2a is, for example, a reflective optical sensor, and is disposed so as to irradiate light above the belt conveyor 2 and detect the well plates 3 by receiving reflected light. The sensor 2a does not have to be optical, and may be contact type. Furthermore, if the sensor 2a is optical, it is not limited to being reflective, but may also be transmissive.

[0026] The reagent storage unit 12 includes a freezer for storing enzyme reagents and a refrigerator for storing primer reagents. The reagent storage unit 12 stores an enzyme reagent container 121 containing an enzyme reagent and a primer reagent container 122 containing a primer reagent. The enzyme reagent contains DNA polymerase, dNTP, and reverse transcriptase as main components. As will be described later, the sample preparation robot 11 prepares the reagent used in preparing a sample set by taking a predetermined amount of reagent from each of the enzyme reagent container 121 and the primer reagent container 122 and dispensing it into a mixed reagent container 16. Hereinafter, a reagent prepared by mixing an enzyme reagent and a primer reagent will be referred to as a "mixed reagent." The enzyme reagent is preferably stored below freezing (for example, -18°C), and the primer reagent is preferably stored at a low temperature of 1°C to 5°C.

[0027] The QC sample storage unit 13 includes, for example, a refrigerator for storing positive and negative controls at low temperatures of 1 to 5°C. The positive control is a specimen containing a known concentration of target nucleic acid or a buffer solution containing a known concentration of target nucleic acid. For example, if the test subject is the SARS-CoV-2 virus, the target nucleic acid is the nucleic acid (RNA or artificially synthesized DNA) or nucleic acid fragment of the SARS-CoV-2 virus. The negative control is a specimen that does not contain the target nucleic acid, such as pure water or a buffer solution.

[0028] The nozzle tip storage unit 14 stores a plurality of nozzle tips that can be attached and detached to the tip of the robot arm of the sample preparation robot 11. The nozzle tips are made of disposable material and are discarded after use by the sample preparation robot.

[0029] The 8-tube storage area 15 is disposed between the sample preparation robot 11 and the measurement robot 22 and holds a plurality of 8-tube storage areas 40 .

[0030] The specimen preparation robot 11 is, for example, a vertical articulated robot. The specimen preparation robot 11 has an end effector 111 at the tip of its arm. The end effector 111 has a syringe pump as a tool, and a nozzle tip stored in a nozzle tip storage unit 14 can be detachably attached to the tip.

[0031] When preparing a mixed reagent, the sample preparation robot 11 uses a nozzle tip attached to the end effector 111 to aspirate a predetermined amount of the enzyme reagent from the enzyme reagent container 121 stored in the reagent storage section 12 and the primer reagent from the primer reagent container 122, and dispenses them into the mixed reagent container 16.

[0032] When dispensing the prepared mixed reagent, the sample preparation robot 11 uses a nozzle tip attached to the end effector 111 to aspirate a predetermined amount of the mixed reagent contained in the mixed reagent container 16 and dispenses it into the container 41 of the 8-tube 40.

[0033] When dispensing a subject sample, the sample preparation robot 11 uses a nozzle tip attached to the end effector 111 to aspirate the subject specimen contained in the well of the well plate 3 transported by the belt conveyor 2 and dispenses it into the container 41 of the 8-tube 40.

[0034] When dispensing a positive control or a negative control, the sample preparation robot 11 uses a nozzle tip attached to the end effector 111 to aspirate a predetermined amount of the positive control and negative control from the QC sample storage section 13 and dispense them into the container 41 of the 8-tube tube 40.

[0035] The sample preparation robot 11 prepares test samples by dispensing and mixing a subject sample and a mixed reagent into the container 41 of the 8-tube array 40. Similarly, the sample preparation robot 11 prepares control samples by dispensing and mixing a positive control or negative control and a mixed reagent into the container 41 of the 8-tube array 40. Because one 8-tube array 40 has eight containers 41, a sample set containing up to eight samples can be prepared. If a sample set includes two types of control samples, the number of test samples will be up to six. As will be described in detail later, in this embodiment, a predetermined amount (enough for 48 tests) of mixed reagent is prepared from a predetermined amount of enzyme reagent aspirated from one enzyme reagent container 121 and a predetermined amount of primer reagent aspirated from one primer reagent container 122. One positive control sample and one negative control sample are prepared each time a mixed reagent is prepared. For example, when a mixed reagent for 48 tests is prepared, the sample set initially prepared includes one positive control sample, one negative control sample, and six test samples.

[0036] The PCR unit group 20 is composed of a plurality of PCR units 21 that can amplify nucleic acids and measure the amplified nucleic acids independently and in parallel. The PCR units 21 are measurement units that amplify nucleic acids contained in a sample set and measure the amplified nucleic acids under the control of a PCR control unit 33. In this embodiment, the measurement robot 22 distributes each sample set (8-tube array 40) to the plurality of PCR units 21 in order according to a predetermined rule. One 8-tube array 40 can be set in one PCR unit 21. One PCR unit 21 can simultaneously perform nucleic acid amplification processing on up to eight samples.

[0037] In this embodiment, the PCR unit group 20 is made up of 16 PCR units 21 (hereinafter sometimes referred to as "PCR units U1 to U16"). Each PCR unit 21 is a measurement unit of the same type, and performs the same processing and measurement.

[0038] The measuring robot 22 is a dual-arm robot and has two robot arms. End effectors 221 and 222 are provided at the tips of the two robot arms, respectively. Robot hands are attached to the end effectors 221 and 222 as tools capable of grasping and moving the octuplet tube 40. Under the control of the robot control unit 32, the measuring robot 22 sets the octuplet tube 40 containing the sample set in the PCR unit 21. After measurement, the measuring robot 22 removes the octuplet tube 40 from the PCR unit 21 and discards it.

[0039] FIG. 2 is a perspective view of the PCR unit 21. The PCR unit 21 has a thermal cycler 23, a tube holder 24 in which an 8-tube strip 40 is set, and an openable cover 25 equipped with an optical unit 26. The PCR unit 21 performs reverse transcription and nucleic acid amplification on a test sample and a control sample, and measures the nucleic acid amplification of each sample. The thermal cycler 23 is a device that heats and cools the 8-tube strip 40 set in the tube holder 24. The reverse transcription process in the PCR unit 21 is performed by heating the sample to a predetermined temperature (e.g., 45°C) for a certain period of time. By heating the sample, the reverse transcriptase contained in the enzyme sample causes a reverse transcription reaction of the nucleic acids in the sample to proceed.

[0040] After the reverse transcription process is completed, the PCR unit 21 starts the nucleic acid amplification process. The nucleic acid amplification process is a process in which a nucleic acid amplification cycle is repeated, in which the sample is heated and cooled at a predetermined interval. For example, the nucleic acid amplification process is performed 45 times, in which heating to 95°C and cooling to 60°C are repeated at 75-second intervals. The thermal cycler 23 is equipped with a Peltier element, and the Peltier element heats and cools the 8-tube strip 40 set in the tube holder 24, thereby performing the nucleic acid amplification process for each sample. The nucleic acids amplified by this process are detected by an optical unit 26 mounted on the cover 25.

[0041] The optical unit 26 includes a light source 261 and a fluorescence detector 262 (see FIG. 4). The light source 261 is disposed facing the 8-tube array 40 set in the tube holder 24 when the cover 25 is closed, and irradiates the samples contained in each container 41 with excitation light. The fluorescence detector 262 detects fluorescence from the sample generated by light irradiation from the light source 261 when the cover 25 is closed. An example of the light source 261 is an LED, and an example of the fluorescence detector 262 is a photodiode. A plurality of light sources 261 and fluorescence detectors 262 are installed on the cover 25, for example, along the direction in which the containers 41 of the 8-tube array 40 are arranged. The method for introducing a fluorescent substance used in PCR measurement into a sample is not particularly limited, and may be, for example, an intercalator method, a probe method, a cycling probe method, or a calcein method. Alternatively, a method for measuring turbidity or absorbance without using a fluorescent substance may be used.

[0042] In PCR measurements, as nucleic acids are amplified through repeated nucleic acid amplification cycles, the fluorescence intensity detected by the fluorescence detector 262 increases. The cycle number at which the fluorescence intensity exceeds a predetermined threshold increases if the sample contains a large amount of the target nucleic acid being tested, and decreases if the target nucleic acid is absent or present in a small amount. Alternatively, if the sample does not contain the target nucleic acid, the fluorescence intensity does not change even as the cycle number increases. Therefore, this cycle number can be used to confirm the presence or absence of the target nucleic acid in the sample. The PCR control unit 33, for example, acquires the fluorescence intensity detected by the fluorescence detector 262 and determines the cycle number at which the fluorescence intensity exceeds the predetermined threshold. The cycle number at which the fluorescence intensity exceeds the predetermined threshold is called the Ct value.

[0043] The PCR control unit 33, for example, creates an amplification curve of fluorescence intensity or an amplification curve of nucleic acid based on fluorescence intensity, compares the amplification curve with a predetermined threshold value set for fluorescence intensity, and calculates the number of cycles of nucleic acid amplification when the fluorescence intensity exceeds the threshold value as the Ct value. The calculation of the Ct value is not limited to a method of comparing fluorescence intensity with a threshold value; instead, the second derivative of the amplification curve may be calculated and the point at which the value is maximum may be used as the Ct value. If the Ct value is less than a predetermined number of cycles, the sample is likely to be positive, and if the Ct value is equal to or greater than the predetermined number of cycles, the sample is likely to be negative. The PCR control unit 33 stores the Ct value in association with information identifying each sample. The PCR control unit 33 may also store the amplification curve together with the Ct value. The identification information for the test sample is, for example, a specimen number assigned to each subject.

[0044] FIG. 3 is a perspective view of an eight-tube tube 40. As shown in FIG. 3, the eight-tube tube 40 is a group of eight containers 41 arranged in a row, with adjacent containers 41 connected to each other to form an integrated group. Because the eight-tube tube 40 has a structure in which eight independent containers 41 are integrated, eight containers 41, each containing eight samples, can be transported simultaneously, contributing to improved throughput. Each container 41 has a cylindrical container body 411 with a bottom that contains a sample, and a lid 412 that closes the opening of the container body 411. The container body 411 and the lid 412 are made of, for example, a translucent resin.

[0045] The sample sets contained in one 8-tube array 40 are prepared using the same mixed reagent dispensed from the same mixed reagent container 16. In this embodiment, multiple sample sets (e.g., six sample sets) contained in multiple 8-tube arrays 40 are prepared using the same mixed reagent. As will be described in detail below, a single mixed reagent container 16 contains an amount of mixed reagent sufficient to prepare 48 test samples that make up the six sample sets, and one QC group made up of the 48 test samples includes at least one positive control sample and one negative control sample.

[0046] 4 is a block diagram showing the configuration of the nucleic acid analyzer 1. As shown in FIG. 4, the belt conveyor 2, the sample preparation robot 11, and the measurement robot 22 are communicatively connected to a robot control unit 32, and their operations are controlled by the robot control unit 32. Each PCR unit 21 constituting the PCR unit group 20 is communicatively connected to a PCR control unit 33, and their operations are controlled by the PCR control unit 33. The PCR control unit 33 acquires the fluorescence intensity detected by the fluorescence detector 262 from each PCR unit 21, creates an amplification curve of the fluorescence intensity, and determines the Ct value. The robot control unit 32 and the PCR control unit 33 are communicatively connected to a system control unit 31.

[0047] The system control unit 31 is a device that controls the nucleic acid analyzer 1 overall and includes a CPU 311, a memory unit 312, a communication interface (IF) 313, a display unit 314, and an input unit 315. The system control unit 31 may be configured as a personal computer. The system control unit 31 executes software stored in the memory unit 312 to comprehensively control the operation of the nucleic acid analyzer 1, and outputs control commands to the robot control unit 32 and the PCR control unit 33 to perform processes related to the preparation of sample sets, distribution of sample sets, PCR measurement, and analysis of measurement results (measurement data). The system control unit 31 is communicably connected to a host computer 50 and transmits analysis results analyzed by the PCR control unit 33 to the host computer 50. The host computer 50 is used, for example, by a doctor to check the analysis results and determine whether each test sample is positive or negative.

[0048] The storage unit 312 is composed of RAM, ROM, a hard disk, etc. The storage unit 312 stores a computer program for executing the above-mentioned processes. This computer program is executed by the CPU 311. The display unit 314 is composed of a display and displays the operating status and analysis results of the nucleic acid analyzer 1. The input unit 315 is composed of a keyboard and mouse and is used to input, for example, identification information of the specimens contained in the well plate 3 and a request for specimen testing. The communication interface 313 includes, for example, an Ethernet-compatible communication module.

[0049] The robot control unit 32 is a device that controls the belt conveyor 2, the sample preparation robot 11, and the measuring robot 22, and includes a CPU 321, a storage unit 322, and a communication interface 323. The robot control unit 32 controls the operations of the belt conveyor 2, the sample preparation robot 11, and the measuring robot 22 based on control commands from the system control unit 31.

[0050] Like the robot control unit 32, the PCR control unit 33 has a CPU 331, a storage unit 332, and a communication interface 333, and controls the operation of each PCR unit 21 that constitutes the PCR unit group 20 based on control commands from the system control unit 31. The PCR control unit 33 may be configured as a personal computer.

[0051] The PCR control unit 33 acquires information about each sample to be distributed to each PCR unit 21 from the system control unit 31 and stores it in the memory unit 332. The information about each sample is identification information for each sample, and is associated with position information for the container 41 of the 8-tube array 40 in which the sample is stored. The identification information for each sample is, for example, a specimen number if the sample is a test sample, or information indicating the type of sample or the lot number of the control sample if the sample is a control sample. As described above, the PCR control unit 33 acquires measurement data (fluorescence intensity detected by the fluorescence detector 262) from each PCR unit 21, creates an amplification curve, and calculates the Ct value. The PCR control unit 33 outputs analysis results that associate the identification information for each sample with the amplification curve and Ct value to the system control unit 31.

[0052] The PCR unit 21 has a thermal cycler 23 including a Peltier element, a light source 261 and a fluorescence detector 262 mounted on a cover 25, a CPU 270, a storage unit 271, and a communication interface 271. The CPU 270 controls the operations of the thermal cycler 23, the light source 261, etc. by executing a program stored in the storage unit 271 in response to a control command transmitted from the PCR control unit 33.

[0053] The thermal cycler 23 heats and cools the 8-tube strip 40 set in the tube holder 24 under the control of the PCR control unit 33 and the CPU 270, thereby performing reverse transcription and nucleic acid amplification. The light source 261 irradiates each container 41 of the 8-tube strip 40 with excitation light under the control of the PCR control unit 33 and the control unit 27. The fluorescence detector 262, under the control of the PCR control unit 33 and the control unit 27, detects the intensity of fluorescence emitted from the sample in each cycle of nucleic acid amplification, and transmits measurement data consisting of time-series data of the fluorescence intensity to the PCR control unit 33.

[0054] An example of an analysis process performed by the nucleic acid analyzer 1 will be described in detail below with reference to Figures 5 to 12. Figures 5 to 8 are flowcharts showing the operation of the nucleic acid analyzer 1, illustrating the cooperation of four control entities that make up the nucleic acid analyzer 1 (a system control unit 31, a robot control unit 32, a PCR control unit 33, and a PCR unit 21).

[0055] 5 is a flowchart showing the operation of the system control unit 31. In step S1, the system control unit 31 determines whether a new well plate 3 has been placed on the belt conveyor 2. The determination in step S1 is YES when the sensor 2a detects the new well plate 3 placed on the belt conveyor 2 and a detection signal from the sensor 2a is input to the system control unit 31 via the robot control unit 32. If the determination in step S1 is YES, the process proceeds to step S2. Note that the determination in step S1 may be YES when the user inputs an inspection request via the input unit 315, instead of detection by the sensor 2a.

[0056] In step S2, the system control unit 31 transmits a start command to the robot control unit 32 to start a series of analysis processes including preparation of a sample set, PCR measurement, and analysis of the measurement data. As will be described in detail later, upon receiving the start command, the robot control unit 32 controls the sample preparation robot 11 to prepare a sample set from the subject specimens in the well plate 3, and sets the 8-tube array 40 containing the sample set in one of the PCR units 21 determined according to a predetermined rule.

[0057] In step S3, the system control unit 31 determines whether or not a setting completion notification has been received from the robot control unit 32. As will be described later, the setting completion notification is a notification that notifies that an 8-tube conduit 40 has been set in the PCR unit 21, and includes identification information U1 to U16 of one PCR unit 21 in which the 8-tube conduit 40 has been set. If a setting completion notification has been received, the system control unit 31 sends a measurement command to the PCR control unit 33 (step S4). The measurement command is a control command for starting PCR measurement in the PCR unit 21 corresponding to the setting completion notification. As will be described in detail later, upon receiving the measurement command, the PCR control unit 33 sends a control command to the specified PCR unit 21 to start PCR measurement, and when the measurement is completed, sends a measurement completion notification to the system control unit 31.

[0058] In step S5, the system control unit 31 determines whether or not a measurement completion notification has been received from the PCR control unit 33. If a measurement completion notification has been received, the system control unit 31 transmits a removal command to the robot control unit 32 to remove the octal tube 40 from the PCR unit 21 (step S6).

[0059] In step S7, the system control unit 31 determines whether or not the analysis results have been received from the PCR control unit 33. If the analysis results have been received, the system control unit 31 outputs the analysis results to the host computer 50 in step S8. The order of steps S6, S7, and S8 may be reversed. If there are multiple host computers 50, the system control unit 31 may select the host computer 50 to which the analysis results are to be sent based on the sample identification information. Furthermore, the analysis results received from the PCR control unit 33 may be combined with additional information and output to a specific host computer 50. Once the system control unit 31 has output the analysis results, it returns to step S1.

[0060] FIG. 6 is a flowchart showing the operation of the robot control unit 32. In step S10, the robot control unit 32 determines whether a start command has been received from the system control unit 31. If the start command has been received, the robot control unit 32 checks whether the mixed reagent necessary for preparing the sample set is present (step S11). If the mixed reagent is present, the process proceeds to step S13. If the mixed reagent is not present, that is, if the number of remaining tests for the prepared mixed reagent is zero, the robot control unit 32 controls the sample preparation robot 11 to prepare the mixed reagent (step S12). Under the control of the robot control unit 32, the sample preparation robot 11 aspirates a predetermined amount of reagent from the enzyme reagent container 121 and the primer reagent container 122 in the reagent storage unit 12, and dispenses the reagent into the mixed reagent container 16 to prepare the mixed reagent. In this embodiment, a mixed reagent for 48 tests is prepared in one mixed reagent preparation.

[0061] In step S13, the robot control unit 32 controls the belt conveyor 2 to move the well plate 3 to a position accessible by the sample preparation robot 11. In step S14, the robot control unit 32 controls the sample preparation robot 11 to prepare test samples from the subject samples contained in each well of the well plate 3 and the mixed reagent contained in the mixed reagent container 16, thereby preparing a sample set. Under the control of the robot control unit 32, the sample preparation robot 11 prepares, for example, a sample set including only the test sample, and a sample set including the test sample and a control sample. In step S14, the sample preparation robot 11 prepares samples by dispensing the subject samples, mixed reagent, etc. into each container 41 of the 8-tube tube 40 in the 8-tube storage area 15, thereby preparing a sample set including up to eight samples. The processing of step S14 will be described later.

[0062] In step S15, the robot control unit 32 determines, according to a predetermined rule, in which of the PCR units 21 U1 to U16 the octopus tube 40 should be set. The robot control unit 32 controls the measurement robot 22 to open the cover 25 of the PCR unit 21 determined as the destination, set the octopus tube 40 in the tube holder 24, and close the cover 25. The predetermined rule is, for example, the order U1, U2, U3, ... U16. When the octopus tube 40 is set in the tube holder 24, for example, a sensor in the PCR unit 21 detects the octopus tube 40, and a detection signal from the sensor is transmitted to the robot control unit 32 and the PCR control unit 33. In step S16, the robot control unit 32 transmits a setting completion notification to the system control unit 31. The robot control unit 32 transmits the setting completion notification, for example, upon receiving the detection signal. As described above, the setting completion notification includes the identification information of the PCR unit in which the octopus tube 40 is set, for example, one of U1 to U16.

[0063] When the 8-tube array 40 is set in the PCR unit 21, the PCR unit 21 starts PCR measurement under the control of the system control unit 31 and the PCR control unit 33.

[0064] The robot control unit 32 determines whether PCR measurement has been completed for each sample in the 8-tube array 40 and whether a removal command has been received from the system control unit 31 (step S17). If a removal command has been received, the robot control unit 32 controls the measurement robot 22 to remove the 8-tube array 40 that has been measured from the PCR unit 21 (step S18). Specifically, under the control of the robot control unit 32, the measurement robot 22 removes the 8-tube array 40 that corresponds to the removal command from the PCR unit 21 and moves it to a predetermined location. The predetermined location may be, for example, a disposal box that stores the 8-tube array 40 that has been measured.

[0065] 7 is a flowchart showing the operation of the PCR control unit 33. In step S20, the PCR control unit 33 determines whether or not a measurement command has been received from the system control unit 31. If a measurement command has been received, the PCR control unit 33 transmits a nucleic acid amplification / detection command to the designated PCR unit 21 corresponding to the measurement command (step S21). As will be described in detail later, upon receiving the nucleic acid amplification / detection command, the PCR unit 21 performs PCR measurement on each sample in the 8-tube array 40 set in the tube holder 24 and transmits the measurement data to the PCR control unit 33. Each sample in the 8-tube array 40 simultaneously undergoes nucleic acid amplification processing and is irradiated with excitation light, and fluorescence generated from each sample is detected.

[0066] The measurement command includes identification information for the target PCR unit and identification information for each sample in the 8-tube strip 40 (sample set) set in the tube holder 24. The identification information for each sample is associated with the number of the container 41 in the 8-tube strip 40, and is stored in the memory unit 332 for use in analyzing the measurement data (see FIG. 10, described below). That is, based on the measurement command, the PCR control unit 33 has information about what sample is contained in which container 41 of the 8-tube strip 40 set in each PCR unit 21. This allows the PCR control unit 33 to output analysis results in which the amplification curves and Ct values obtained from the measurement results for each sample in each PCR unit 21 are associated with the identification information for each sample.

[0067] In step S22, the PCR control unit 33 determines whether or not measurement data has been received from the PCR unit 21. If measurement data has been received, the PCR control unit 33 sends a measurement completion notification to the system control unit 31 (step S23). As described above, upon receiving the measurement completion notification, the system control unit 31 sends a take-out command to the robot control unit 32 (step S6 in FIG. 5). The PCR control unit 33 analyzes the received measurement data in step S24 to create analysis results, and sends the analysis results to the system control unit 31 in step S25. The measurement data is analyzed according to the procedure shown in FIG. 9, which will be described later. The analysis results are sent to the host computer 50 via the system control unit 31 (step S8 in FIG. 5) and are used by a doctor to determine whether the result is positive or negative.

[0068] FIG. 8 is a flowchart showing the operation of the PCR unit 21. In step S30, the PCR unit 21 determines whether or not a nucleic acid amplification / detection command has been received from the PCR control unit 33. If a nucleic acid amplification / detection command has been received, the PCR unit 21 performs nucleic acid amplification processing and fluorescence detection in step S31. Specifically, the PCR unit 21 heats the octuplet tube 40 using the thermal cycler 23 to perform a reverse transcription process, and then performs nucleic acid amplification processing by repeating a nucleic acid amplification cycle of heating and cooling the octuplet tube 40 at a predetermined period. In this embodiment, a nucleic acid amplification cycle of heating to 95°C and cooling to 60°C is performed 45 times. Furthermore, in each nucleic acid amplification cycle, the PCR unit 21 irradiates each container 41 of the octuplet tube 40 with excitation light using the light source 261, and detects fluorescence emitted from the sample using the fluorescence detector 262.

[0069] Fluorescence detection is performed at a specific timing in the nucleic acid amplification cycle. For example, when the sample is cooled to 60°C, excitation light is irradiated and the intensity of fluorescence emitted from the sample is measured. The PCR unit 21 measures the fluorescence intensity in each nucleic acid amplification cycle for all samples contained in the 8-tube array 40. In step S32, the PCR unit 21 transmits this measurement data to the PCR control unit 33. The measurement data may be transmitted each time a nucleic acid amplification cycle is completed, or may be transmitted collectively after all nucleic acid amplification cycles are completed and 45 measurements have been performed.

[0070] FIG. 9 is a flowchart showing the procedure for analyzing measurement data in the PCR control unit 33.

[0071] In step S240, the PCR control unit 33 calculates the Ct value of the test sample based on the measurement data of the test sample from the measurement data corresponding to one sample set received in step S22. Specifically, the PCR control unit 33 creates an amplification curve of fluorescence intensity based on the measurement data, and calculates the Ct value based on the cycle number at which the fluorescence intensity exceeds a threshold. The amplification curve and Ct value are stored in the memory unit 332 of the PCR control unit 33. The threshold is set, for example, to a signal level at which a significant increase is observed in the measured fluorescence intensity relative to the baseline signal, and is stored in advance in the memory unit 332.

[0072] In step S241, the PCR control unit 33 determines whether the received measurement data includes measurement data of a control sample. If measurement data of a control sample is not included, the processing of FIG. 9 ends and the process returns to the main routine. If measurement data of a control sample is included, the PCR control unit 33 calculates the Ct value of the control sample based on the measurement data of the control sample, in the same manner as described in step S240 (step S242).

[0073] In step S243, the PCR control unit 33 performs quality control determination based on the Ct value of the control sample calculated in step S242.

[0074] The quality control determination for the positive control sample is performed by checking whether the Ct value of the positive control sample is equal to or less than a first cycle number (e.g., 32 cycles) set as a threshold. If the Ct value is equal to or less than the first cycle number, the sample is determined to be "normal." If the Ct value exceeds the first cycle number, the sample is determined to be "abnormal." The first cycle number is the minimum cycle number that the positive control sample should exhibit if there are no abnormalities in the reagent quality and the series of processes including the sample preparation step and nucleic acid amplification step, and is pre-stored in the memory unit 332 of the PCR controller 33. If the Ct value of the positive control sample exceeds the first cycle number, this indicates that the rise in fluorescence intensity is slower than normal, and an abnormality such as an inability to prepare a reagent of the desired quality or a malfunction of the nucleic acid amplification cycle may be suspected.

[0075] The negative control sample is also subjected to quality control assessment in the same manner as the positive control sample. Quality control assessment for the negative control sample is performed by checking whether the Ct value of the negative control sample is equal to or greater than a second cycle number (e.g., 40 cycles) set as a threshold. If the Ct value is equal to or greater than the second cycle number, the sample is determined to be "normal." If the Ct value is smaller than the second cycle number, the sample is determined to be "abnormal." The second cycle number is a cycle number that the negative control sample would not exhibit if there were no abnormalities in the series of processes, and is pre-stored in the memory unit 332 of the PCR control unit 33. If the Ct value of the negative control sample is smaller than the second cycle number, this means that the rise in fluorescence intensity is faster than the standard, and there is a possibility that the negative control sample contains target nucleic acid that should not be present. In this case, contamination is suspected.

[0076] If the quality control judgment for both the positive control sample and the negative control sample is "normal," the PCR control unit 33 sets an "OK" flag for the analysis results of the same QC group prepared using the same mixed reagent as the control sample (step S244).On the other hand, if the quality control judgment for either the positive control sample or the negative control sample is "abnormal," the PCR control unit 33 sets an "NG" flag for the analysis results of the same QC group prepared using the same mixed reagent as the control sample (step S245).

[0077] 10 is a schematic diagram of an analysis result database 1000 (hereinafter simply referred to as DB) stored in the memory unit 332 of the PCR control unit 33. DB 1000 is created in the memory unit 332 of the PCR control unit 33. DB 1000 includes a column C1 in which the specimen number is stored, a column C2 in which the specimen type is stored, a column C3 in which the identification information of the PCR unit used for nucleic acid analysis is stored, a column C4 in which the well number in which the sample is contained is stored, a column C5 in which the Ct value is stored, a column C6 in which the quality control determination results are stored, and a column C7 in which the identification information of the reagents used is stored.

[0078] The information in columns C1 to C4 is included in the measurement command that the PCR control unit 33 received from the system control unit 31 in step S20. For the Ct value in column C5, the Ct value of the test sample obtained in step S240 is stored for the test sample record. For the control sample record, the Ct value of the control sample obtained in step S242 is stored. Column C6 stores either an "OK" or "NG" flag as the result of the judgment made by the PCR control unit 33 based on the Ct value of the control sample in step S243.

[0079] In the example of Figure 10, sample set A contains positive control sample P1001, negative control sample N1001, and six test samples S1001-S1006. Sample set B contains eight test samples S1039-S1046. Sample set A and sample set B constitute QC group X, which were prepared using the same mixed reagent R1. In the example of Figure 10, the quality control judgment results for the positive control sample P1001 and negative control sample N1001 in sample set A are both OK. In this case, "OK" is stored as the quality control judgment result for the six test samples in sample set A, which is the same as positive control sample P1001 and negative control sample N1001. Furthermore, "OK" is also stored as the quality control judgment result for test samples S1039-S1046 in sample set B, which is also included in the same QC group X. Although not shown in the figure, "OK" is also stored as the quality control judgment result for test samples in other sample sets besides sample sets A and B in the same QC group X.

[0080] If the quality control judgment for both or either of the positive control sample P1001 and the negative control sample N1001 is "abnormal," "NG" is stored as the quality control judgment result for all test samples S1001 to S1046 included in the same QC group X, including the test samples in sample sets A and B (step S245). If the quality control judgment result is NG, information indicating that retesting is required is added to the analysis result when it is output from the PCR control unit 33 to the system control unit 31. As such, in this embodiment, based on the measurement results of the control sample included in one sample set (sample set A in FIG. 10), quality control judgment is performed for the test samples (S1001 to S1006) included in one sample set and the test samples (S1039 to S1046) included in another sample set (sample set B in FIG. 10).

[0081] According to this embodiment, the number of control samples measured can be significantly reduced compared to quality control performed based on conventional concepts (see Figure 21, described below), thereby reducing reagent costs and improving throughput. Furthermore, even if the number of control samples measured is reduced, at least one positive control sample and one negative control sample are measured in the same QC group using a common mixed reagent, so reagent accuracy can be guaranteed. More specifically, if the measurement results of the positive control sample are valid, it can be confirmed that the mixed reagent was properly prepared and that the enzymes and primers contained in the mixed reagent are properly amplifying nucleic acids. Furthermore, if the measurement results of the negative control sample are valid, it can be confirmed that at least the mixed reagent is free of contamination. In other words, the nucleic acid analysis method of this embodiment reduces reagent costs and improves throughput for measurement of control samples, while ensuring the quality of the mixed reagent—specifically, the appropriateness of its preparation and the presence or absence of contamination—and enabling nucleic acid analysis of test samples.

[0082] The analytical accuracy of a testing system equipped with a nucleic acid analyzer 1 is significantly affected by the mixed reagent used to prepare samples. In other words, if there is a problem with the preparation of the mixed reagent or with the enzyme and primer reagents used as raw materials, and the mixed reagent is not of the desired quality, accurate measurement data will not be obtained. If an abnormality is detected in the quality control assessment, it is highly likely that the desired mixed reagent was not prepared. Therefore, in this embodiment, all samples in the same QC group are considered to have an abnormality in analytical accuracy. While the measurement accuracy of the PCR unit 21 is guaranteed by various sensors installed in the unit, it is also preferable to periodically measure control samples in each unit, as shown in the modified example described below.

[0083] In addition, when a quality control determination is made that there is an abnormality, if measurement of a test sample of the same QC group has not been completed in another PCR unit U2 to U6, the PCR control unit 33 may cancel that measurement. In this case, unnecessary measurements can be eliminated, leading to improved throughput. In the example shown in Figure 9, if a quality control determination is made that there is an abnormality and the word "NG" is displayed in the analysis result column for a test sample of the same sample group, data analysis of that test sample is not performed, but data analysis may also be performed on the test sample displayed as NG, and the Ct value may be calculated.

[0084] In the example of Figure 10, DB1000 includes only the Ct value of the test sample in the analysis result record, but it may also store a positive / negative judgment result for each sample as a result of comparing the Ct value of the test sample with a predetermined threshold. A Ct value below the threshold can be judged as positive, and a Ct value above the threshold can be judged as negative. Note that the quality control judgment results stored in column C6 of Figure 10 are not limited to OK / NG, as long as they can distinguish whether or not there is a quality control abnormality. For example, they may be "Good / Bad" or "Pass / Fail," or the absence of a quality control abnormality may be indicated by "0" and the presence of an abnormality by "1."

[0085] Fig. 11 is a diagram showing an example of sample arrangement (hereinafter referred to as "Example 1") in the PCR unit group 20. In Fig. 11, "P" represents a positive control sample, "N" represents a negative control sample, and a blank circle represents a test sample.

[0086] In Example 1, a positive control sample and a negative control sample are measured for each mixed reagent prepared at one time, and the measurement results of the control sample are applied to the measurement results of multiple sample sets measured in the PCR unit 21 that measures the control sample and another PCR unit 21. As described above, the sample preparation unit mixes the enzyme reagent and the primer reagent to prepare multiple mixed reagents (e.g., enough for 48 tests) in the mixed reagent container 16. At least one sample set including a control sample is prepared each time a new mixed reagent is prepared. In Example 1, 48 test samples are prepared using the same mixed reagent. In other words, six sample sets, each containing eight samples, are prepared.

[0087] The positive control sample and the negative control sample are included in the first sample set, which is the first of six sample sets included in the same QC group (the hatched area in Figure 11) prepared using the same mixed reagent. The first sample set is the first of the six sample sets to be transported to PCR unit 21 and measurement to begin. In this case, quality control judgment based on the measurement data of the control sample is performed first, so the analysis results of the test sample (subject sample) can be returned immediately after measurement of the first sample set is completed, thereby shortening the turnaround time (TAT) from test request to result report.

[0088] In Example 1, a set of six samples from the first QC group prepared from the same mixed reagent is measured in PCR units U1 to U6, and a set of six samples from the second QC group is measured in PCR units U7 to U12. A set of six samples from the third QC group is measured in PCR units U13 to U16, and U1 and U2. By the time the measurement of the sample set from the third QC group begins, the measurement of the sample set from the first QC group in PCR units U1 and U2 has been completed and the octet tube 40 has been removed, so the sample set from the third QC group can be transported to PCR units U1 and U2.

[0089] By using multiple PCR units 21, each measuring up to eight samples in parallel, the waiting time for samples to be collected is shortened compared to conventional analytical devices that batch process large quantities of samples using well plates, significantly reducing the turnaround time for collected samples. Each sample set is distributed to the multiple PCR units 21 in sequence according to a predetermined rule. In Example 1, the sample sets are distributed in the order of their creation: PCR units U1, U2, U3, U16, U1, U2, etc. Regularly distributing each sample set to the multiple PCR units U1 to U16 enables efficient processing and measurement. In Example 1, the operation of the device is controlled so that the measurement of the first sample set in the first QC group is completed and the removal of the 8-tube strip 40 is completed by the time the fifth sample set in the third QC group is transported to PCR unit U1.

[0090] In Example 1, as described above, one positive control sample and one negative control sample are prepared for each QC group. Furthermore, the positive and negative control samples are included only in the first sample set prepared first in each QC group, and are not included in the other sample sets prepared second through sixth. In Example 1, the first and second sample sets are prepared at a ratio of 1:5. In this case, 46 of the 48 samples that make up one QC group are test samples.

[0091] FIG. 21 is a diagram showing sample arrangement based on the conventional concept. Based on the conventional concept, as shown in FIG. 21, a sample set including one positive control sample and one negative control sample is measured in each PCR unit. In this case, 1 / 4 of the total measurements are measurements of control samples. Therefore, if the conventional concept is applied to the nucleic acid analyzer 1, throughput will decrease and reagent costs will increase. In contrast, according to the method of Example 1, 46 out of 48 measurements are measurements of subject samples, and measurements of control samples are reduced to 1 / 24 of the total measurements. Therefore, according to the method of Example 1, it is possible to achieve a significant reduction in reagent costs and an improvement in throughput compared to methods based on the conventional concept.

[0092] FIG. 12 is a flowchart showing the sample set preparation process corresponding to Example 1. In Example 1, as described above, six sample sets are prepared as one QC group prepared from the same mixed reagent. The six sample sets are prepared by the sample preparation robot 11 under the control of the robot control unit 32. In step S140, the robot control unit 32 determines whether the sample set to be prepared is the first sample set prepared in the QC group. Hereinafter, the sample set prepared first in the QC group will be referred to as the head sample set. If the control sample set is the head sample set (YES in step S140), the sample preparation robot 11 dispenses the positive control sample and the negative control sample into the containers 41 of the 8-tube array 40, respectively, under the control of the robot control unit 32.

[0093] Next, the sample preparation robot 11 dispenses the subject samples from the wells of the well plate 3 into the remaining six containers 41 of the eight-tube array 40 (step S142). Next, the sample preparation robot 11 dispenses the mixed reagent into each container 41 of the eight-tube array 40 to prepare control samples and test samples (step S143). Through the above process, a sample set including two control samples and six test samples is prepared. Note that in this embodiment, the subject samples and the mixed reagent are dispensed into the eight-tube array in this order, but the order may be reversed.

[0094] If it is determined in step S140 that the sample set to be prepared is not the first sample set, i.e., if the sample set to be prepared is the second or subsequent sample set in the QC group, the control sample preparation process in step S141 is skipped and the process proceeds to step S142. That is, the sample preparation robot 11 dispenses the subject's sample into all of the first to eighth containers 41 of the 8-tube array 40 to prepare a sample set including eight test samples.

[0095] 13 is a diagram showing another example (hereinafter referred to as "Example 2") of sample arrangement in the PCR unit group 20. Differences from Example 1 will be described in detail below.

[0096] Example 2 is similar to Example 1 in that six sample sets in the same QC group each contain one positive control sample and one negative control sample, and the measurement results of the control sample are applied to the measurement results of multiple sample sets measured in a PCR unit 21 that measures the control sample and another PCR unit 21. However, Example 2 differs from Example 1 in that the positive control sample and the negative control sample are included in different sample sets. In Example 2, of the six sample sets that make up a QC group, the first sample set prepared first contains the negative control sample, and the last sample set prepared contains the positive control sample.

[0097] In Example 2, the first sample set includes a negative control sample, the second to fifth sample sets include only test samples, and the final sample set includes a positive control sample. These samples are then transported to PCR units U1 to U6 in this order for measurement. This reduces the risk of contamination of the negative control sample or test sample with the positive control. The negative control sample is preferably prepared before the test sample in the first sample set, and is contained in the first container 41 of the eight-tube array 40. The positive control sample is preferably prepared after the test sample in the final sample set, and is contained in the eighth container 41 of the eight-tube array 40.

[0098] In Example 2, sample sets containing control samples (first and last sample sets) and sample sets containing only test samples are prepared in a ratio of 1:2. However, since the first and last sample sets each contain one control sample, in this case too, 46 of the 48 test samples that make up one QC group are test samples.

[0099] FIG. 14 is a flowchart showing the sample set preparation process of Example 2. In step S144, the robot control unit 32 determines whether the sample set to be prepared is the head sample set of six sample sets constituting the same QC group (step S144). If it is the head sample set (YES in step S144), the robot control unit 32 controls the sample preparation robot 11 to dispense the negative control sample and the subject sample into each container 41 of the 8-tube strip 40 (step S145). In step S145, under the control of the robot control unit 32, the sample preparation robot 11 dispenses the negative control sample into the first container 41 of the 8-tube strip 40, and then dispenses the subject sample from the wells of the well plate 3 into the second to eighth containers 41 of the 8-tube strip 40. Next, the sample preparation robot 11 dispenses the mixed reagent from the mixed reagent container 16 into each container 41 of the 8-tube strip 40 to prepare the negative control sample and the test sample (step S146).

[0100] If the answer is NO in step S144, the robot control unit 32 then determines whether the sample set to be prepared is the last of the six sample sets constituting the same QC group (step S147). If the robot control unit 32 determines that it is the last sample set (YES in step S147), the robot control unit 32 controls the sample preparation robot 11 to dispense the positive control sample and the subject sample into each container 41 of the 8-tube array 40 (step S148). In step S148, after the subject sample is dispensed into the first to seventh containers 41 of the 8-tube array 40, the positive sample is dispensed into the eighth container 41.

[0101] If the sample set to be prepared is neither the first nor the last sample set, i.e., if the second to fifth sample sets in the QC group are to be prepared, the subject's sample is dispensed into each container 41 of the 8-tube array 40 (step S149). After steps S148 and S149 are completed, the process proceeds to step S146, where the mixed reagent is dispensed.

[0102] FIG. 15 is a diagram showing another example of sample arrangement in the PCR unit group 20 (hereinafter referred to as "Example 3").

[0103] Example 3 is similar to Example 2 in that multiple sample sets constituting the same QC group contain one positive control sample and one negative control sample, and the positive control sample and the negative control sample are contained in separate sample sets. In Example 2, the positive control sample was contained in the last sample set prepared among the sample sets of the same QC group. In other words, the positive control sample is contained in the last sample set that can be prepared with one mixed reagent. On the other hand, Example 3 takes into consideration the case where a test request is interrupted and it takes time for the next subject sample to be supplied. In Example 3, even if there is remaining mixed reagent sufficient to prepare more samples than the maximum number (8) of samples included in the sample set, a sample set containing a positive control sample is prepared at the time the test request is interrupted.

[0104] In Figure 15, arrows A and B indicate the time when test requests are interrupted, i.e., the time when the supply of well plates 3 containing subject samples is temporarily stopped. At the time of arrow A, the sample preparation unit is preparing the fourth sample set constituting the same QC group, and although there is more than eight test amounts of mixed reagent remaining, the fourth sample set contains a positive control sample. Similarly, at the time of arrow B, there is more than eight test amounts of mixed reagent remaining, but the fifth sample set contains a positive control sample. When well plates 3 containing subject samples are continuously supplied to the nucleic acid analyzer 1 without interruption, a positive control sample is prepared in the last sample set of the same QC group, as in Example 2.

[0105] In this way, if the supply of subject samples is interrupted, the sample set being prepared will include a positive control sample even if it is not the last sample set of the same QC group.If the supply of subject samples is interrupted and it will take some time to prepare the last sample set, the positive control sample can be measured first, so that the measurement results can be quickly applied to the analysis of test samples that have already been measured.According to Example 3, when the supply of subject samples to the nucleic acid analyzer 1 is intermittent, the TAT can be significantly reduced compared to Example 2.

[0106] In Example 3, when the supply of subject samples resumes, a sample set is prepared using the remaining mixed reagent. If a sample set is prepared with empty containers 41 in the 8-tube array 40 when the supply of subject samples is interrupted, seven or more sample sets are prepared for one QC group. Note that since the positive control sample has already been prepared, there is no need to prepare a positive control sample for the last sample set of the same QC group. If the measurement of the positive control sample has been completed, the analysis results of the test samples can be returned as soon as the measurement of the remaining sample sets is completed.

[0107] In Example 3, the following four types of sample sets can be prepared as sample sets of the same QC group: In Example 3, the following sample set (2) is prepared only if the supply of subject samples is interrupted during the preparation of the sample set. (1) A sample set including a negative control sample and a test sample (2) A sample set including a negative control sample, a positive control sample, and a test sample. (3) A sample set including a positive control sample and a test sample. (4) A sample set containing only test samples

[0108] Among sample sets of the same QC group, the sample set (1) above is the first sample set to be prepared when the number of supplied subject samples is greater than the number of containers 41 of the 8-tube strip 40 that contains the sample set. The sample set (2) above is the first sample set to be prepared when one or more containers 41 of the 8-tube strip 40 remain after dispensing the negative control and all supplied subject samples. The sample set (3) above is the second or subsequent sample set to be prepared when the positive control has not been dispensed and the remaining amount of mixed reagent is equivalent to one sample, or when the positive control has not been dispensed and one or more containers 41 of the 8-tube strip 40 remain after dispensing all supplied subject samples. The sample set (4) above is prepared when the sample set preparation conditions (1) to (3) are not met.

[0109] According to Examples 1 to 3, one positive control sample and one negative control sample are prepared for each QC group, and quality control of test samples belonging to the same QC group is performed based on the measurement results of the positive control sample and the negative control sample. If there are no problems with the measurement results of the positive control sample and the negative control sample, it can be confirmed that there are no abnormalities due to improper preparation of the mixed reagent or contamination in test samples of the same QC group prepared using the same mixed reagent. Therefore, according to Example 1, nucleic acid analysis of test samples is possible while checking for the presence or absence of abnormalities due to improper preparation of the mixed reagent or contamination based on the positive control sample and negative control sample contained in one QC group.

[0110] FIG. 16 is a diagram showing another example of sample arrangement in the PCR unit group 20 (hereinafter referred to as "Example 4").

[0111] Example 4 is similar to Example 1 in that the first sample set prepared from six sample sets in the same QC group contains one positive control sample and one negative control sample. However, Example 4 differs from Example 1 in that the other sample sets besides the first sample set also contain negative control samples. In the first sample set of Example 4, the first container 41 of the eight-tube strip 40 contains a positive control sample, and the second container 41 contains a negative control sample, but as in Example 1, the order of the control samples may be reversed.

[0112] In Example 4, in quality control based on positive control samples, the measurement results of the positive control sample included in the first sample set are applied to the quality control judgment of the test samples in the same QC group. Meanwhile, for negative control samples, the quality control judgment of the test samples included in each sample set is performed based on the measurement results of the negative control samples included in each sample set. For example, in the example of FIG. 16, sample set A measured by Run 1 of PCR unit 1 includes a positive control sample and a negative control sample, and sample set B measured by Run 1 of PCR unit 2 includes a negative control sample. In this case, if the quality control judgment results of the positive control sample and the negative control sample included in sample set A are both OK, the quality control judgment results of the six test samples included in sample set A will also be OK. For the test samples included in sample set B, if the quality control judgment of the positive control sample included in sample set A is OK and the quality control judgment of the negative control sample included in sample set B is OK, the quality control judgment result will be OK.

[0113] According to Example 4, each QC group contains one positive control sample and one negative control sample, so that nucleic acid analysis of test samples can be performed while confirming that there are no abnormalities due to improper preparation of the mixed reagent or contamination, as in Examples 1 to 3. In Example 4, each sample set also contains a negative control sample, so that contamination during the preparation of each sample set can also be detected.

[0114] FIG. 17 is a diagram showing another example of sample arrangement in the PCR unit group 20 (hereinafter referred to as "Example 5").

[0115] Example 5 is similar to Examples 1 and 4 in that the first sample set prepared from six sample sets in the same QC group contains one positive control sample and one negative control sample. However, Example 5 differs from Examples 1 and 4 in that the other sample sets besides the first sample set also contain positive control samples. In the first sample set of Example 5, the positive control sample is contained in the first container 41 of the 8-tube array 40, and the negative control sample is contained in the second container 41. However, as in Example 1, the order of the control samples may be reversed. The concentration of the positive control in the positive control sample of the second sample set may be different from the concentration of the positive control sample in the first sample set. For example, it may be the concentration of the detection sensitivity guaranteed by the PCR unit 21. Alternatively, it may be a concentration within ±1 to 10% of the detection sensitivity concentration.

[0116] According to Example 5, each QC group contains one positive control sample and one negative control sample, so that nucleic acid analysis of test samples can be performed while confirming that there are no abnormalities due to improper preparation of the mixed reagent or contamination, as in Examples 1 to 3. In Example 5, each sample set also contains a positive control sample, so that it can be confirmed that each PCR unit 21 is operating normally and that nucleic acid amplification and detection of amplified nucleic acid are being performed normally.

[0117] In Examples 4 and 5, in the second sample set, the control sample is contained in the first container 41 of the 8-tube array 40, but it may be contained in another container 41. The ratio of the first and second sample sets is 1:5, but it may be 1:n (n≧2) (the same applies to other Examples). Also, although all of the second sample sets contain control samples, some of the second sample sets may contain only test samples.

[0118] 18 to 20 are diagrams showing modified examples of the arrangement of control samples in the PCR unit group 20. 18 to 20 show only the arrangement of control samples in PCR units U1 to U16. "NP" stands for both negative and positive control samples.

[0119] In the example shown in FIG. 18, control samples are periodically measured in specific PCR units U1 to U16 from the first measurement (run 1) to the ninth measurement (run 9). The arrangement of these control samples is the same as in Example 1. Specifically, in PCR units U1, U3, U5, U7, U9, U11, U13, and U15, control samples are measured once every four measurements. On the other hand, in PCR units U2, U4, U6, U8, U10, U12, U14, and U16, no control sample measurements were performed until the ninth measurement. Therefore, if any of the PCR units U1 to U16 has not measured a control sample within a predetermined time or period, it is preferable to automatically measure a control sample in that unit and periodically check the detection accuracy of that unit.

[0120] 18, the control sample is measured in the tenth measurement of PCR units U2, U4, U6, U8, U10, U12, U14, and U16. In this case, control sample measurement is always performed at least once per predetermined period (e.g., once every 10 measurement cycles) in all PCR units U1 to U16. For example, the robot control unit 32 counts the number of measurement cycles that do not include control sample measurement for each PCR unit U1 to U16, and if there is a unit for which this number of measurement cycles exceeds a predetermined threshold, it controls each robot 11, 22 to transport a sample set including the control sample to that unit, where the control sample is measured.

[0121] In the example shown in Figure 19, similar to the example shown in Figure 18, control sample measurements are performed in all PCR units U1 to U16, for example, once every 10 measurement cycles. In PCR units U2, U6, and U10, no control sample measurements are performed until the ninth measurement, but if the above-mentioned predetermined threshold is set to nine, control sample measurements are performed in PCR units U2, U6, and U10 in the tenth measurement. In the example shown in Figure 19, a sample set containing only positive control samples and a sample set containing only negative control samples are transported to the PCR units as control samples, and a sample set containing both positive and negative control samples is also transported to the PCR units.

[0122] By the ninth measurement, a sample set including a negative control sample has been transported to PCR units U12 and U14, and the negative control sample has been measured in PCR units U12 and U14, but no positive control sample has been measured. In this case, a sample set including a positive control sample can be transported to PCR units U12 and U14 in the tenth measurement, and the positive control sample can be measured. For example, robot controller 32 counts the number of measurement cycles that do not include measurements of positive and negative control samples for each PCR unit U1-U16 and for each type of control sample. If there is a unit where the number of measurement cycles that do not include measurements of at least one control sample exceeds a predetermined threshold, robot controller 32 controls each robot 11, 22 to transport a sample set including an unmeasured control sample to that unit, and then measure that control sample.

[0123] In the example shown in FIG. 20, when the frequency of measurement of the same type of control sample increases in a particular PCR unit 21, the PCR unit 21 that measures that control sample is changed. That is, the order of the PCR units 21 (the PCR units 21 to which the sample set is distributed) is changed to distribute the destinations of the control samples. For example, in PCR unit U3, a negative control sample is measured in the third measurement cycle, and measurement of the negative control sample is scheduled to continue in the fourth measurement cycle. Similarly, in PCR unit U4, a positive control sample is measured in the first measurement cycle, and measurement of the positive control sample is scheduled again in the fourth measurement cycle, resulting in a high frequency of measurement of the positive control sample. In such a case, the PCR unit 21 that measures the control sample is changed from the PCR unit 21 that measures the control sample more frequently to the PCR unit 21 that measures the control sample less frequently.

[0124] In the example shown in Figure 20, a sample set including a negative control sample that was scheduled to be distributed to PCR unit U3 in the third measurement cycle is distributed to PCR unit U4. Furthermore, a sample set including a positive control sample that was scheduled to be distributed to PCR unit U4 in the fourth measurement cycle is distributed to PCR unit U7. Because the positive control sample has just been measured in PCR units U5 and U6, this sample set is distributed to PCR unit U7. When a sample set including two control samples is scheduled to be distributed, as in the third measurement cycle of PCR unit U11, a sample set including a positive control sample and a sample set including a negative control sample may be prepared instead of this sample set. Because the positive control sample was measured in PCR unit U11 in the first measurement cycle, the sample set including the positive control sample of these two sample sets is distributed to a unit other than PCR unit U11.

[0125] The modified examples shown in Figures 18 to 20 prevent the measurement frequency of control samples from increasing in a particular PCR unit 21, making it easier to periodically check the detection accuracy of each PCR unit 21. For example, the robot controller 32 counts the measurement frequencies of positive and negative control samples for each PCR unit U1 to U16 and for each type of control sample. If a unit has a control sample measurement frequency exceeding a predetermined threshold, the robot controller 32 controls each robot 11, 22 to transport the sample set containing the frequently measured control sample to another unit. The other unit is a unit whose control sample measurement frequency is equal to or less than a predetermined threshold, and is determined, for example, based on a predetermined measurement order. Alternatively, a unit with a low control sample measurement frequency may be preferentially selected as the other unit, as long as it does not interfere with overall measurements.

[0126] As described above, the nucleic acid analysis method of the above-described embodiment and modified example is a completely new and innovative method in which the measurement results of a control sample measured in one PCR unit 21 are also used in the analysis of a test sample measured in another PCR unit 21. The measurement results of the control sample are applied to the analysis of a test sample prepared from the same reagents as the control sample, which is measured in another PCR unit 21, for example. In this case, the number of control sample measurements can be significantly reduced. This reduces the amount of reagent used, significantly reducing reagent costs, and also increases the proportion of test sample measurements in the total measurements, significantly improving throughput. By appropriately setting the scope of application of the measurement results of the control sample, accurate quality control can be performed even when the number of control sample measurements is reduced.

[0127] The present invention is not limited to the above-described embodiments and modifications, and can be appropriately modified without impairing the object of the present invention. For example, although the nucleic acid analyzer 1 is configured to dispense positive and negative controls to prepare a control reagent, the positive and negative controls may be supplied from the upstream side of the testing system, similar to the case of subject samples. The positive and negative controls may be supplied to the nucleic acid analyzer 1 by the belt conveyor 2 while contained in wells of the well plate 3.

[0128] In the above-described embodiment, a method was exemplified in which at least one positive control sample and one negative control sample were measured each time a new mixed reagent prepared by mixing an enzyme reagent and a primer reagent was used. However, the positive control sample and the negative control sample may also be measured each time a new manufacturing lot of a reagent (e.g., an enzyme reagent or at least one primer reagent) is used. For example, at least one sample set including a positive control sample and a negative control sample is prepared each time a new manufacturing lot of a reagent is used. In other words, even if a new mixed reagent is used, as long as the manufacturing lot of the reagents from which the mixed reagent is made remains the same, each sample prepared from reagents from the same manufacturing lot may be treated as one QC group, and one positive control sample and one negative control sample may be measured per group.

[0129] In the above-described embodiment, eight samples contained in the 8-tube array 40 constitute one sample set. However, the form is not limited as long as multiple samples contained in multiple connected containers constitute one sample set. For example, a maximum of 96 samples contained in a 96-well plate may constitute one sample set. In this case, in a preferred embodiment, the PCR unit may be configured to accept the 96-well plate and perform at least nucleic acid amplification on the 96 samples in a batch process. In addition to nucleic acid amplification, the PCR unit may also perform measurement of the amplified nucleic acid.

[0130] Furthermore, the reagent used to prepare the sample may be of one type. For example, when a predetermined container is used to contain the amount of reagent used to prepare multiple test samples, at least one sample set including a positive control sample and a negative control sample may be prepared each time the container is changed. As in Example 2, when the positive control sample and the negative control sample are contained in different sample sets, at least one sample set including a positive control sample and one sample set including a negative control sample are prepared each time the container is changed or each time the manufacturing lot of the reagent is changed.

[0131] In the above-described embodiment, an example was shown in which multiple PCR units capable of independently amplifying nucleic acids and detecting amplified nucleic acids were used to perform nucleic acid analysis in parallel. Each PCR unit was equipped with a thermal cycler, a light source, and a fluorescence detector. In another example of the present invention, nucleic acid amplification and detection of amplified nucleic acids may be performed by separate devices. For example, multiple thermal cycler modules capable of independently amplifying nucleic acids may be used. In this case, the thermal cycler unit does not need to be equipped with a light source and a fluorescence detector for detecting nucleic acids.

[0132] 22 and 23 are schematic diagrams showing another embodiment, in which an 8-tube tube 40 is set in a thermal cycler module 200 (hereinafter simply referred to as module 200).

[0133] Figure 22 is a schematic diagram showing the overall configuration of a nucleic acid analyzer 1' according to a modified example. In Figure 22, the same components as those in Figure 1 are given the same reference numerals, and detailed description thereof will be omitted. In the modified example of Figure 22, a measurement robot 220 includes an end effector 221 consisting of a hand for setting an eight-tube array 40 containing a sample into a module 200, and an end effector 224 consisting of a hand for loading the module 200, in which the eight-tube array 40 has been set, into a detection device 80.

[0134] As shown in FIG. 23 , the detection device 80 is installed on, for example, a table 70. The detection device 80 has a disk-shaped turntable 90 that can rotate around a central column 100, and a plurality of, for example, eight, module mounting portions 91 arranged on the turntable 90. Each module mounting portion 91 extends radially from the center of the turntable 90. A module 200 can be mounted on each module mounting portion 91 with its longitudinal direction facing the radial direction. The module mounting portions 91 are arranged at equal intervals in the circumferential direction R of a circle centered at the center of the turntable 90. That is, the eight module mounting portions 91 are arranged at 45-degree intervals in the circumferential direction R.

[0135] The detection device 80 includes an optical detector 120. The optical detector 120 has a shape that extends radially outward from the center of the turntable 90, and corresponds to one module mounting section 91 (a module 10 of the module mounting section 91). When the module mounting section 91 is positioned downward, the optical detector 120 can detect fluorescence associated with nucleic acid amplification of multiple samples contained in multiple containers of a series of tubes 40 in the modules 200 of the module mounting section 91. In other words, the optical detector 120 can detect fluorescence associated with nucleic acid amplification of multiple samples on a module-by-module basis, and is shared by the eight modules 200.

[0136] The optical detector 120 has a light source unit and a photodetector unit. The optical detector 120 can irradiate each container of the 8-tube array 40 with light from the light source unit, and detect the fluorescence of the nucleic acid in the sample generated by the light using the photodetector unit. The light source unit includes a plurality of light-emitting elements, such as LEDs, arranged in a row along the radial direction of the turntable 90. The photodetector unit, like the light source unit, includes a plurality of light-receiving elements, such as photodiodes, arranged in a row along the radial direction of the turntable 90. One light-emitting element and one light-receiving element are arranged in pairs, and the fluorescence generated by the light irradiated by the light-emitting element is detected by the paired photodetector.

[0137] The module 200 includes a lid 201 provided with a plurality of holes corresponding to each container of the 8-tube array 40, a main body 202 in which the 8-tube array 40 is placed, a thermal cycler 203, and a CPU 204 provided inside the main body 202. The module 200 can be placed on the module mounting section 91.

[0138] The measurement robot 223 grasps the module 200 with the arm 224 and sets it on the module mounting section 91. When the module 200 is set on the module mounting section 91, the turntable 70 rotates. With the module 200 set on the module mounting section 91, the CPU 204 controls the thermal cycler 203 to heat and cool the 8-tube array 40 to amplify nucleic acids. The CPU is synchronized with the nucleic acid analyzer 1, and one cycle of the module 200 rotating around the turntable 90 is synchronized with one cycle of nucleic acid amplification consisting of heating and cooling. The nucleic acid analyzer 1 controls the turntable 90 so that the module 200 is positioned directly below the fluorescence detector 120 at the timing of fluorescence detection in each cycle of nucleic acid amplification. This allows nucleic acids amplified by multiple modules 200 to be detected by a single fluorescence detector 120.

[0139] In this modification, nucleic acid amplification can be performed by individual modules 200, and amplified nucleic acids can be detected using a common fluorescence detector 120. In this modification as well, a first sample set including at least one control sample and a second sample set not including at least one of the control samples included in the first sample set can be prepared by the sample preparation robot 11. The prepared sample sets can be subjected to nucleic acid amplification and amplified nucleic acids can be detected by the modules 200 and the detection device 80. [Explanation of symbols]

[0140] 1 Nucleic acid analyzer 2 conveyor belts 3-well plate 11. Sample preparation robot 12 Reagent Storage Section 13 QC Sample Storage Department 14 Nozzle tip storage area 15 8-tube storage area 16 Mixing reagent container 20 PCR units 21, U1~U16 PCR units 22 Measuring Robot 23 Thermal Cycler 24 Tube holding part 25 Cover 26 Optics Department 261 Light source 262 Fluorescence Detector 27 Control Unit 31 System control section 311 CPU 312 Storage section 313 Communication Interface 314 Display section 315 Input section 32 Robot control unit 321 CPU 322 Storage section 323 Communication Interface 33 PCR control section 331 CPU 332 Storage section 333 Communication Interface 40 8-tube 41 Container 411 Container body 412 Lid 50 Host Computer

Claims

1. preparing a first sample set including a test sample prepared from a first subject specimen and a reagent, a positive control and a positive control sample prepared from the reagent, and a negative control and a negative control sample prepared from the reagent; creating a second sample set comprising a test sample prepared from a specimen from a second subject and the reagent, and comprising one or both of the positive control sample and the negative control sample included in the first sample set; performing nucleic acid amplification on the first sample set and measuring the amplified nucleic acids; performing nucleic acid amplification on the second sample set and measuring the amplified nucleic acids; analyzing the measurement results of each of the test samples contained in the first and second sample sets based on the measurement results of the positive control sample and the negative control sample contained in at least the first sample set; The nucleic acid amplification of the first sample set is performed by a first unit that receives a sample set and performs nucleic acid amplification; A nucleic acid analysis method, wherein nucleic acid amplification for the second sample set is performed by a second unit that receives one sample set and performs nucleic acid amplification.

2. The method according to claim 1 , wherein nucleic acid amplification by the first unit and nucleic acid amplification by the second unit are performed in parallel.

3. the first and second units each include a detector for detecting the amplified nucleic acid; measuring the amplified nucleic acids of the first sample set by the detector of the first unit; The method of claim 1 or 2, wherein the measurement of the amplified nucleic acids of the second sample set is performed by the detector of the second unit.

4. The method according to claim 3 , wherein the measurement of the amplified nucleic acid by the first unit and the measurement of the amplified nucleic acid by the second unit are carried out in parallel.

5. The method according to any one of claims 1 to 4, wherein the first and second sample sets are a plurality of samples contained in a plurality of linked containers.

6. The method of any one of claims 1 to 5, wherein the first sample set comprises at least one positive control sample, one negative control sample, and a plurality of test samples.

7. The method according to any one of claims 1 to 6, wherein the test samples included in the first sample set and the test samples included in the second sample set are each a plurality of samples prepared from a plurality of subject specimens.

8. The method of any one of claims 1 to 7, wherein the second sample set includes one of the positive control sample and the negative control sample included in the first sample set.

9. The method of any one of claims 1 to 7, wherein the second sample set does not include both the positive control sample and the negative control sample included in the first sample set.

10. The method according to any one of claims 1 to 9, wherein the first and second sample sets are prepared using a common reagent at a ratio of 1:n (n is an integer of 2 or more).

11. The method according to any one of claims 1 to 10, wherein the first sample set is prepared as at least one of a plurality of sample sets prepared using the common reagent.

12. The method further includes preparing a mixed reagent as the reagent by mixing a first reagent and a second reagent; The method according to any one of claims 1 to 11, wherein at least one first sample set is prepared each time the mixed reagent is switched.

13. The method according to any one of claims 1 to 12, wherein at least one first sample set is prepared each time a manufacturing lot of the reagent used is changed.

14. preparing a first sample set including a test sample prepared from a first subject specimen and a reagent, a positive control and a positive control sample prepared from the reagent; creating a second sample set comprising a second subject specimen and a test sample prepared from the reagent; preparing a third sample set including a third subject specimen and a test sample prepared from the reagent, a negative control and a negative control sample prepared from the reagent; the second sample set does not include the positive control sample and the negative control sample; performing nucleic acid amplification on the first sample set and measuring the amplified nucleic acids; performing nucleic acid amplification on the second sample set and measuring the amplified nucleic acids; performing nucleic acid amplification on the third sample set and measuring the amplified nucleic acids; A nucleic acid analysis method for analyzing the measurement results of each test sample contained in the first to third sample sets based on the measurement results of the positive control sample contained in the first sample set and the measurement results of the negative control sample contained in the third sample set.

15. preparing a first sample set including a test sample prepared from a first subject specimen and a reagent, a positive control and a positive control sample prepared from the reagent; preparing a second sample set including a second subject specimen and a test sample prepared from the reagent, a negative control and a negative control sample prepared from the reagent; performing nucleic acid amplification on the first sample set and measuring the amplified nucleic acids; performing nucleic acid amplification on the second sample set and measuring the amplified nucleic acids; A nucleic acid analysis method further comprising analyzing the measurement results of the test samples contained in the first and second sample sets based on the measurement results of the positive control sample contained in the first sample set and the measurement results of the negative control sample contained in the second sample set.

16. preparing a first sample set including a test sample prepared from a first subject specimen and a reagent, a negative control sample prepared from the reagent, and a negative control sample; preparing a second sample set including a second subject specimen and a test sample prepared from the reagent, a positive control and a positive control sample prepared from the reagent; performing nucleic acid amplification on the first sample set and measuring the amplified nucleic acids; performing nucleic acid amplification on the second sample set and measuring the amplified nucleic acids; A nucleic acid analysis method further comprising analyzing the measurement results of the test samples contained in the first and second sample sets based on the measurement results of the negative control sample contained in the first sample set and the measurement results of the positive control sample contained in the second sample set.

17. Distributing the first and second sample sets into a plurality of units capable of amplifying nucleic acids individually for each sample set; performing nucleic acid amplification on the sample set distributed in each of the plurality of units; and The nucleic acid analysis method according to any one of claims 1 to 16, further comprising measuring the amplified nucleic acid.

18. 18. The method of claim 17, wherein the first and second sample sets are distributed sequentially among the units according to a predetermined rule.

19. 18. The method of claim 17, wherein the first sample set is distributed to units of the plurality of units that meet a predetermined condition.

20. 20. The method of claim 19, wherein the predetermined condition is that the control sample has not been measured within a predetermined time or period.

21. a sample preparation device for preparing a plurality of sample sets, each of which includes a test sample prepared from a subject specimen and a reagent; a unit for amplifying nucleic acids for each of the plurality of sample sets and measuring the amplified nucleic acids; A control unit; Equipped with The sample preparation device, under the control of the control unit, prepares a first sample set including the test sample, a positive control and a positive control sample prepared from the reagent, and a negative control and a negative control sample prepared from the reagent, and a second sample set including the test sample and including either the positive control sample or the negative control sample included in the first sample set, or excluding both; The unit includes a first unit that performs nucleic acid amplification on the first sample set and a second unit that performs nucleic acid amplification on the second sample set; The control unit analyzes the measurement results of each test sample contained in the first and second sample sets based on the measurement results of at least the positive control sample and the negative control sample contained in the first sample set.

22. a sample preparation device for preparing a plurality of sample sets, each of which includes a test sample prepared from a subject specimen and a reagent; a unit for amplifying nucleic acids for each of the plurality of sample sets and measuring the amplified nucleic acids; A control unit; Equipped with The sample preparation device, under the control of the control unit, prepares a first sample set including the test sample, a positive control, and a positive control sample prepared from the reagent, a second sample set including the test sample, and a third sample set including the test sample, a negative control, and a negative control sample prepared from the reagent, wherein the second sample set does not include the positive and negative control samples; the units include a first unit that performs nucleic acid amplification on the first sample set, a second unit that performs nucleic acid amplification on the second sample set, and a third unit that performs nucleic acid amplification on the third sample set; The control unit analyzes the measurement results of each test sample contained in the first to third sample sets based on the measurement results of the positive control sample contained in the first sample set and the negative control sample contained in the third sample set.

23. a sample preparation device for preparing a plurality of sample sets, each of which includes a test sample prepared from a subject specimen and a reagent; a unit for amplifying nucleic acids for each of the plurality of sample sets and measuring the amplified nucleic acids; A control unit; Equipped with The sample preparation device prepares, under the control of the control unit, a first sample set including the test sample, a positive control, and a positive control sample prepared from the reagent, and a second sample set including the test sample, a negative control, and a negative control sample prepared from the reagent; The unit includes a first unit that performs nucleic acid amplification on the first sample set and a second unit that performs nucleic acid amplification on the second sample set; The control unit analyzes the measurement results of each test sample contained in the first and second sample sets based on the measurement results of the positive control sample contained in the first sample set and the negative control sample contained in the second sample set.

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