JPH09159671A - Method and equipment for measuring components derived from microorganism - Google Patents

Abstract

PROBLEM TO BE SOLVED: To measure microorganisms with high reliability over a wide range through easy measuring operation by providing means for holding cuvets containing a sample liquid to be inspected, a light emitting diode for irradiating the sample cuvet with a light beam, and means for detecting the quantity of transmitted light. SOLUTION: The inventive equipment comprises sample cuvets 1 for containing a sample liquid to be inspected, a diode light source 2, emitting light of blue and/or violet, and a photoelectric detector 3 for detecting the quantity of light emitted from the light source 2 and transmitted through the cuvet 1. The inventive equipment further comprises an incubator 4 for sustaining the cuvet 1 at a constant temperature in stationary state, and a multiplexer 5 for transmitting a signal while switching the quantity of transmitted light. A computer 8 is interlocked with the detection of cuvet 1 being set in a sample holding means to start measuring of time of one unoperating cuvet and a timer 10 measures the reaction time thus controlling the processing of data indicative of variation in the quantity of transmitted light and general operation of equipment.

Description

Detailed Description of the Invention

[0001]

BACKGROUND OF THE INVENTION 1. Field of the Invention The present invention relates to an apparatus and method for measuring a microorganism-derived component, and more specifically, to a wide range of microbial contamination such as Gram-negative bacteria, Gram-positive bacteria and fungi with high reliability and ease. The present invention relates to an apparatus and a method for measuring a microbial-derived component that can be measured by various measurement operations.

[0002]

2. Description of the Related Art Manufacturing and quality control of pharmaceuticals, manufacturing and quality control of medical equipment, manufacturing and quality control of food, quality control of dialysate, diagnosis of microbial infections, impurity control of washing water in semiconductor manufacturing process In areas where microbial contamination such as
Conventionally, the amount of microbial contamination has been measured to prevent its contamination. Here, the microbial contamination amount is more specifically the number of viable bacteria in a sample, or endotoxin, peptidoglycan, (1 → 3) β-D-glucan (hereinafter, referred to as β-glucan), etc. It is the amount of components derived from microorganisms. If the amount of microbial contamination is measured based on the amount of components derived from microorganisms, not only the amount of viable bacteria can be estimated, but also the total amount of microbial contamination including dead bacteria can be known.

Recently, the following method is becoming mainstream as a method for measuring the amount of components derived from microorganisms, because it is easy to operate because breeding of experimental animals and cell culture are unnecessary. Measurement of endotoxin derived from Gram-negative bacteria by the so-called Limulus test (nephelometric analysis) using Limulus test (blood cell component of horseshoe crab). Measurement of endotoxin derived from Gram-negative bacteria by Limulus test (synthetic substrate method). Β derived from fungus by Rimurus test (turbidimetric analysis)
-Measurement of glucan Component that reacts with β-glucan extracted from horseshoe crab hemocyte components and measurement of fungal β-glucan by a reagent containing a synthetic substrate (synthetic substrate method) SLP reagent (a reagent containing a silkworm body fluid component) Of peptidoglycan derived from Gram-positive bacteria or Gram-negative bacteria or β-glucan derived from fungi by activation time analysis method)

The turbidimetric time analysis method, the activation time analysis method, and the synthetic substrate method are the following methods, respectively. Turbidity time analysis method or activation time analysis method: The mixture of the sample and the reaction reagent (test sample solution) is irradiated with light, and after the predetermined time has elapsed after mixing the sample and the reaction reagent, the test sample solution Optical change [ratio between initial transmitted light amount Io and transmitted light amount (It) after a predetermined time (t) has elapsed. R = It / Io, logarithmic value of transmitted light amount ratio R, change in transmittance, change in absorbance, etc.] is measured to measure a time (time until the transmitted light amount changes at a certain ratio). , A method of performing the target measurement using the time. Synthetic substrate method: A method in which a synthetic substrate having the property of releasing a dye by the action of an enzyme activated in a test sample solution is used and the target measurement is performed from the amount of the released dye.

The measurement by the turbidimetric time analysis method and the synthetic substrate method in the Limulus test is properly used according to the influence of the target substance as a sample on the reaction. Peptidoglycan derived from Gram-positive bacteria, which was difficult to detect in the conventional Limulus test, etc., can be detected by the development of the above SLP reagent. The law is becoming more and more important.

[0006]

The conventional turbidimetric time analyzer is easy to operate, and by using this,
However, it has the advantage that it can perform highly reproducible and quantitative measurements with high reliability for multiple samples, but it cannot be used for measurements based on the synthetic substrate method such as the above. There wasn't.

In addition, the conventional microplate reader
It was possible to perform the above, etc., but, for example, the reaction start time and the measurement start time could not be linked, and the reaction elapsed time for each test sample solution could not be strictly measured, and the sample container In addition, it is difficult to completely remove microbial contamination of the 96-well microplate, which is a reaction vessel, and to uniformly control the temperature of the test sample solution in the peripheral portion and the central portion of the microplate. Therefore, the obtained measured value has a problem that it is not reliable.

In addition to the above-mentioned problems, in the microplate reader, since it is not possible to carry out the measurement while the microplate is stationary, gelation caused by using the Limulus test as described above is caused. When the analysis method using the reaction is carried out using a microplate reader, there is also a problem that the measurement accuracy deteriorates. Furthermore, the conventional microplate reader is used as a light source.
Since the tungsten lamp is used, a light distribution system is required and a mechanism for moving the microplate is also required, so that the device is complicated and expensive.

The inventions according to claims 1 and 11 are an apparatus and a method for measuring a microorganism-derived component, respectively, which can be applied to both the turbidimetric time analysis method and the synthetic substrate method. An object of the present invention is to provide a measuring device and a measuring method for a microorganism-derived component capable of measuring a wide range of microbial contamination such as Gram-positive bacteria and fungi with high reliability and easy measurement operation. Further, in addition to the above object, an object of the present invention is to provide a measuring device capable of automatically detecting the time when the sample cuvette is installed in the cuvette holding means. In addition to the object of claim 1, the inventions of claims 6 and 7 aim to provide a measuring device in which the stability of the measurement circuit is improved and the measurement accuracy is improved.

[0010]

The measuring apparatus according to claim 1 of the present invention which meets the above object, comprises mixing a sample and a reaction reagent, and measuring the change in the amount of transmitted light due to the start of the reaction after the mixing, thereby deriving from a microorganism. In an apparatus for measuring components, a means for holding a sample cuvette containing a mixture of the sample and a reaction reagent (test sample solution), and blue and / or purple light for irradiating the sample cuvette with light rays are provided. A light emitting diode for emitting light, means for detecting the amount of transmitted light of the irradiated light, and means for detecting the time from the start of measurement until the amount of transmitted light of the test sample changes by a predetermined ratio. To do.

Further, the measuring method according to the eleventh aspect of the present invention is such that the amount of transmitted light changes from a predetermined time after preparation of a mixture of a sample and a reaction reagent (test sample solution) to a predetermined ratio change. In the method for measuring a microorganism-derived component based on the above, the test sample solution is housed in a sample cuvette and held in a stationary state, and a blue and / or purple ray is emitted from the light emitting diode to the test sample cuvette. And detecting the amount of transmitted light of the irradiated light, and detecting the time from the passage of the predetermined time until the amount of transmitted light of the test sample liquid changes by a predetermined ratio,
It is characterized in that the object is measured based on the time.

In summary, the present invention uses a light emitting diode that emits blue and / or violet light to improve the conventional turbidimetric time analyzer (toxinometer), and to analyze the turbidimetric time analysis method and synthesis. The gist is that it can be applied to both the substrate method, but conventionally, there is no measuring device and method for the turbidimetric time analysis method applicable to both the turbidimetric time analysis method and the synthetic substrate method. unknown. The inventors of the present invention have conducted extensive studies to achieve the above-mentioned object, and as a result, in the conventional turbidimetric time analysis method using a light-emitting diode, p at a wavelength of 405 nm which has been conventionally used as a color-forming synthetic substrate reagent.
-The color development of nitroaniline (pNA) could not be detected, but by using a light emitting diode that emits blue and / or violet light, conventional microplates were used.
The present invention has been achieved by discovering that quantitative measurement equivalent to or more than the measurement by a dagger can be carried out.

[0013]

BEST MODE FOR CARRYING OUT THE INVENTION In the present invention, as a light source,
It is characterized by using a light emitting diode that emits blue and / or purple light, but conventionally, this light emitting diode has not been used at all for the measurement of components derived from microorganisms. Such an idea is not known at all. As a light emitting diode for emitting blue and / or violet light, at least one of the intersections of the half height of the maximum peak of light emission and the emission spectrum is in the range of 415 to 485 nm, particularly in the range of 415 to 485 nm. A light emitting diode having a light emitting peak is preferably used.

As the means for detecting the time from the start of measurement of the present invention until the amount of transmitted light of the sample to be tested changes at a constant rate, specifically, for example, the time of installation of the sample cuvette in the holding means is detected. Means, means for starting timing of the timer in synchronization with detection at the time of installation, and means for sampling and storing the amount of transmitted light of the test sample at an arbitrary timing in synchronization with the timer that has started timing. There is a means of doing.

In consideration of measurement accuracy, it is desirable that the means for holding the sample cuvette of the measuring apparatus of the present invention should be capable of holding the sample cuvette in a stationary state.
Further, in consideration of measurement efficiency, it is desirable that a plurality of sample cuvettes can be held. The means for detecting the amount of transmitted light of the irradiated light and the means for detecting the time when the sample cuvette is installed in the holding means are installed for each sample cuvette, a plurality of timers are installed, and a means for starting timing is provided for the sample cuvette. It is advisable to provide a mechanism for starting time measurement of one of the timers that are not operating among the plurality of timers in association with the detection at the time of installation. Further, there are a plurality of means for sampling and storing the transmitted light amount of the sample to be tested at an arbitrary timing in conjunction with the timer that has started the time measurement, and a means for detecting the time from the start of measurement until the transmitted light amount changes at a fixed rate. You may prepare for each.

Next, preferred embodiments of the apparatus of the present invention will be described with reference to the drawings. FIG. 1 is a block diagram of a measuring apparatus according to the present invention, in which a plurality of test sample cuvettes 1 containing a mixture of a sample and a reaction reagent (test sample solution) and a plurality of blue and / or purple lights are provided. A light emitting diode light source 2 which emits light, a photoelectric detector 3 which irradiates a plurality of corresponding sample cuvettes 1 with light rays to detect a plurality of transmitted light amounts separately, and a constant temperature of the sample cuvette 1 in a stationary state The incubator 4 which holds the state, the multiplexer 5 which sequentially switches the plurality of transmitted light amounts detected by the photoelectric detector 3 and transmits the same, and the amplifier circuit 6 which amplifies the transmitted signal.
And an A / D converter 7 for converting the signal amplified by the amplifier circuit 6 into a digital value, and one that is not operating in conjunction with the detection when the sample cuvette 1 is installed in the sample holding means. The timer 8 is started by the timer 8 and measures the reaction time, and the timer that has started the timer is interlocked with to display the time (that is, the measurement) state corresponding to the optical system in the transmitted light measurement state. Element 9 and a timer that starts timing of the digitized transmitted light amount, sampled at an arbitrary timing and stored in the memory 11, and control data processing of change in transmitted light amount and operation of the entire apparatus. An example of the computer 8 is shown.

A glass test tube is preferably used as the sample cuvette 1, and by using a glass test tube, microbial contamination can be completely removed by dry heat treatment. Examples of the light emitting diode light source 2 that emits blue and / or violet light include, for example, the light emitting diode NLPB500, 510, 520, commercially available from Nichia Corporation.
Or NLPB300, 310, 320, Toyoda Gosei Co., Ltd.
Commercially available light emitting devices such as E1L51, E1L53, and E1L55 can be used.

The light emitting diode light source 2 used in the above embodiment is the above NLPB520, which emits blue and violet light and has a light emitting peak at 450 nm, and has a height half that of the light emitting peak. The intersection with the spectrum is 415 nm and 48
It is a 5 nm diode. In the above-mentioned embodiment, a plurality of light sources 2 are provided, but this is not necessarily the case, and the irradiation light may be guided to each test sample cuvette 1 from a single light source by an optical fiber. As the photoelectric detector 3, it is possible to use a light receiving element such as a photo diode or a photoelectric cell that generates an electric signal corresponding to the amount of transmitted light. Similarly to the light source, the photoelectric detector may guide the transmitted light of each test sample cuvette 1 to a single photoelectric detector by an optical fiber.

The incubator 4 for holding the sample cuvette 1 in a constant temperature in a stationary state is formed by forming the means for holding the cuvette 1 by an aluminum block and connecting a known temperature control circuit and elements to it. Good to do. By forming in this way, the incubator 4 having a uniform temperature distribution without vibration can be obtained. If the light source 2 and the photoelectric detector 3 are held together in a constant temperature state in the present incubator 4, the stability of the measuring circuit is further improved. In this device, the sample cuvette 1 can be kept in a stationary state during the measurement. This is an essential condition for carrying out the turbidimetric time analysis method using the Limulus test with high accuracy.

The detection of the time when the sample cuvette 1 is installed in the cuvette holding means can be realized by installing a microswitch or an optical sensor for detecting each cuvette in the cuvette holding means. Is preferably used to detect a rapid change in the amount of transmitted light accompanying the installation of the sample cuvette. By doing this, it is possible to automatically detect the installation time without using extra detection parts, strictly control the reaction elapsed time for each independent sample cuvette, and operate the sample cuvette. Is simply inserted into the holding holder,
It will be extremely easy.

The plurality of timers 10 for starting timed counting each time a sample cuvette is set can be composed of a known sequential circuit and counter, but one basic timer and a computer.
It can be replaced by a program operation by the data 8 and the memory 11. An LED can be used for the element 9 that indicates which optical system is currently in the measuring state, and it is possible to know from the display state which optical system is to enter the measuring state next.

The computer 8 detects the time when the cuvette is installed, determines and controls which of the timers 10 is started, and controls the timing while observing the count of the timer 10 during the time counting operation. The transmitted light amount of the test sample is sampled and stored in the memory 11 as data. It also controls the stop of the timer after a predetermined time.
Since the reaction time is strictly controlled in association with the detection at the time of setting the sample cuvette 1 in this way, the time from the start of measurement to the change in the transmitted light amount of the test sample by a certain ratio is calculated by the computer. 8 and the memory 11 can be easily detected by a program operation. In addition, as a method of confirming that "the amount of transmitted light has changed by a certain ratio", for example, changes in the ratio R of transmitted light, changes in the logarithmic value of the ratio R of transmitted light, changes in transmittance, changes in absorbance, etc. There is a method of checking based on the above. Further, by using the data stored in the memory 11, it is possible to strictly measure the reaction by the end point method or the dynamic reaction by the rate method.

In order to fully exhibit the function of the measuring apparatus of the present invention having the above-mentioned structure, in the above embodiment, the data display device 12 (LED, CRT display, liquid crystal display, etc.) and data display control are provided. A switch 13, a print printer 14 for data, and a communication device 15 for communicating with an external computer are provided. When a CRT display or a liquid crystal display is used for the display device 12, the measurement state display element 9 can be replaced with these displays. Also, an auxiliary storage device 17 such as a floppy disk drive can be installed.

FIG. 2 is a perspective view of a concretely constructed measuring apparatus of the present invention. 12 is a display device for displaying measured sample data, and 13 is a plurality of display data. , A control switch 16 for switching the sample cuvette holder with a temperature controller for holding the sample cuvette to be tested, 9
Is an element that indicates which optical system is currently in the measurement state. Here, as an example, an example is shown in which eight test sample cuvettes are simultaneously held. When the installation of the sample cuvette, which is a mixture of the sample and the reaction reagent, in the holder 16 is detected, one of the timers that are not operating is activated in conjunction with this. The corresponding display element 9 lights up in synchronization with the activation of this timer, and continues to light up until the end of measurement.
If the display element is left unlit, the measurement and data processing are automatically completed, so the measurement operation is extremely easy. In FIG. 2, 14 is a data printing printer and 17 is an auxiliary storage device.

In order to measure the concentration of the microorganism-derived component by the method of the present invention, after the reaction reagent and the test solution are mixed, the time until the amount of the product of the reaction reagent reaches a certain value is permeated. It is detected as the time until the amount of light changes by a certain ratio, and using this, it is quantified by a conventional method (for example, a method using a calibration curve showing the relationship between this time and the amount of microorganism-derived components). Further, in the present invention, the time until the amount of transmitted light changes by a certain ratio is specifically, for example, the change of optical measurement values such as the ratio R of transmitted light, the logarithmic value of R, the transmittance and the absorbance. It is the time until the amount reaches a predetermined value.
The predetermined value is not particularly limited as long as it is a value at which the desired measurement can be carried out. For example, if the transmitted light amount ratio R is used as an optical measurement value, the predetermined value is usually -3 to It is appropriately set within the range of -25%, preferably -5 to -20%. Also, when the measurement of the present invention is carried out by utilizing the transmittance, the absorbance, the logarithmic value of R and the like as the optical measurement value, the predetermined value may be appropriately set according to the case of R.

As the reaction reagent, a known reaction reagent that specifically reacts with a microorganism-derived component may be used. For example, for the measurement of Gram-negative bacteria, known reaction reagents that react with endotoxin, known reaction reagents that react with peptidoglycan, etc. are used, and for Gram-positive bacteria, known reaction reagents that react with peptidoglycan are used, Known reaction reagents that react with β-glucan are used for fungi.
The main components contained in these reaction reagents are contained in the body fluid of insects such as horseshoe crab blood cells and silkworms.

Since the horseshoe crab hemocyte component contains a component that reacts with endotoxin and a component that reacts with β-glucan, it can be used as it is or depending on the purpose.
Only components that react only with endotoxin or β-glucan are used. Since the silkworm body fluid component (SLP) contains a component that reacts with β-glucan and a component that reacts with peptidoglycan, the silkworm body fluid component is used as it is or after being appropriately separated according to the purpose. In the turbidimetric time analysis method, the above-mentioned reaction reagent is used, but in the synthetic substrate method, a reagent in which a color-developing synthetic substrate is further added to the above-mentioned reaction reagent is used. These reaction reagents may be self-made based on known methods or may be prepared by appropriately using commercially available products.

Next, an example of measuring the concentration of a microorganism-derived component using the apparatus of the present invention will be described. Example 1: Turbidity time analysis method In a glass test tube for measuring endotoxin, 100 microliters of an endotoxin aqueous solution (0 to 100 EU / milliliter) and a LAL (hemoglobin blood cell component) solution (Limulus ES-II, Wako Pure Chemical Industries, Ltd.) 100 μl) was added and mixed with stirring, and the change in the amount of transmitted light was measured using the apparatus of the present invention. In FIG. 3, [It (amount of transmitted light from the sample solution at time t) / Io (amount of transmitted light from the sample solution before the decrease started due to the progress of the reaction)] × 1
The calibration relationship between the time (gelation time) required for 00 (%) to reach 95.0% and the endotoxin concentration is shown. As is clear from FIG. 3, a good calibration relationship was obtained in a very wide range.

Into a glass test tube for measuring endotoxin, an endotoxin aqueous solution (0.05 EU / ml) 1
00 microliters and LAL (Liquid blood clot component)
100 microliters of the solution (Limulus ES-J Test Wako, manufactured by Wako Pure Chemical Industries, Ltd.) was added and mixed with stirring, and the reproducibility was measured using the apparatus of the present invention. Table 1
2 shows the reproducibility of the time (gelation time) required until [It / Io] × 100 (%) becomes 95.0% when the endotoxin concentration is 0.05 EU / ml.
For comparison, when a conventional red LED was used as a light source, the same measurement was performed, and the results are also shown in Table 1.

[0030]

[Table 1]

As is clear from Table 1, the reproducibility obtained by the device of the present invention is comparable to that of the conventional device, and
The apparatus of the present invention has the advantage that the gelation time is shorter than that of the conventional apparatus, and the time required for detection can be shortened.

Example 2: Synthetic Substrate Method In a glass test tube for measuring endotoxin, 100 microliters of an endotoxin aqueous solution (0 to 100 EU / milliliter) and a synthetic substrate method reagent solution (Toxicara ^TM System LS-200, Seikagaku Corporation). 100 μl) was added and mixed with stirring, and the change in the amount of transmitted light was measured using the apparatus of the present invention. In FIG. 4, [It / Io] × 1
The calibration relationship between the time (activation time) required for 00 (%) to reach 95.0% and the endotoxin concentration is shown. As is clear from FIG. 4, a good calibration relationship was obtained in a very wide range.

Example 3 β-Glucan Measurement (Synthetic Substrate Method) A cardan (β
-Glucan) 100 microliters of an aqueous solution (0 to 1000 pg / milliliter) and 100 microliters of Bj-Star-A kit color reagent (manufactured by Maruha Co., Ltd.) were added and mixed with stirring, using the apparatus of the present invention. The change in the amount of transmitted light was measured. In FIG. 5, [It / Io] × 100
The calibration relationship between the time required for (%) to reach 90.0% (activation time) and the cardran concentration is shown. As is clear from FIG. 5, a good calibration relationship was obtained in a very wide range.

Example 4: Measurement of SLP reagent In a glass test tube for measuring endotoxin, 100 microliters of a cardan aqueous solution (0 to 1000 pg / ml) and 100 SLP reagent (manufactured by Wako Pure Chemical Industries, Ltd.) were used.
Microliter was added and mixed with stirring, and the change in the amount of transmitted light was measured using the apparatus of the present invention. In FIG. 6, [It /
The relationship between the time required for Io] × 100 (%) to reach 95.0% (activation time) and the cardran concentration is shown. As is clear from FIG. 6, a good calibration relationship was obtained in a very wide range.

[0035]

EFFECTS OF THE INVENTION Conventionally, in order to determine the presence or absence of microbial contamination, a microplate reader has been used to measure the components derived from microorganisms, with insufficient reaction time management and uneven reaction temperature control. However, according to the invention described in claims 1 and 11 of the present invention, the turbidimetric time analyzer is improved. However, the synthetic substrate method, which could not be measured by the conventional turbidimetric time analyzer, can also be measured, so the measurement for determining the presence or absence of microbial contamination of Gram-negative bacteria, fungi, Gram-positive bacteria, etc. can be performed easily. In addition, it has an epoch-making effect that can be performed with high reliability and efficiency, which is not found at all in the conventional apparatus and method for measuring a microorganism-derived component of this kind.

In addition to the effect of claim 1, the invention according to claim 5 can automatically detect the installation time of the sample cuvette without using an extra detection component, and can provide an independent sample. The reaction time for each cuvette can be strictly controlled,
Moreover, there is an advantage that the operation is easy. In addition to the effects of the first aspect, the inventions of the sixth and seventh aspects improve the stability of the measurement circuit and significantly improve the measurement accuracy.

[0037]

[Brief description of the drawings]

FIG. 1 is a block diagram of a measuring apparatus of the present invention.

FIG. 2 is a perspective view of the measuring device of the present invention.

FIG. 3 is a calibration relationship diagram between gelling time and endotoxin concentration by turbidimetric time analysis.

FIG. 4 is a calibration relationship diagram of activation time and endotoxin concentration by the synthetic substrate method.

FIG. 5 is a calibration relationship diagram between activation time for β-glucan measurement and cardran (β-glucan) concentration.

FIG. 6 is a calibration relationship diagram between activation time and cardan concentration using an SLP reagent.

[Explanation of symbols]

1 sample cuvette 2 light emitting diode emitting blue and / or purple light 3 photoelectric detector 4 incubator 5 multiplexer 7 A / D converter 8 computer 9 measurement status display element 10 timer 11 memory 16 sample cuvette Holder

 ─────────────────────────────────────────────────── ─── Continuation of the front page (72) Inventor Ryuzo Tsujino 1-2-28 Kitsurenhigashi, Hirano-ku, Osaka City, Osaka Prefecture

Claims (12)

[Claims] 1. An apparatus for measuring a microorganism-derived component by mixing a sample and a reaction reagent and measuring a change in transmitted light amount due to the start of reaction after mixing, wherein a mixture of the sample and the reaction reagent (test sample) is used. (Sample liquid) means for holding a sample cuvette, a light emitting diode for emitting blue and / or violet light for irradiating the sample cuvette with a light beam, means for detecting the amount of transmitted light of the irradiated light beam, and measurement A device for measuring a microorganism-derived component, comprising: means for detecting a time from the start until the amount of transmitted light of a test sample changes by a predetermined ratio. 2. The means for detecting the time from the start of the measurement until the amount of transmitted light of the sample to be tested changes by a predetermined ratio, the means for detecting the time when the sample cuvette is installed in the holding means, and the time when the installation is performed. 2. The method according to claim 1, further comprising: a means for starting the timekeeping of the timer in association with the detection of the time, and a means for interlocking with the timer that has started the timekeeping to sample and store the amount of transmitted light of the test sample at an arbitrary timing. The measuring device described. 3. The measuring device according to claim 1, wherein the means for holding the sample cuvette is capable of holding a plurality of sample cuvettes in a stationary state. 4. The emission diode for emitting blue and / or violet light has a wavelength of 415 nm to 4 at least at one of the intersections of the half height of the emission peak and the emission spectrum.
The measuring device according to claim 1, which is in a range of 85 nm. 5. The method according to claim 2, wherein the detecting means at the time of setting uses the light emitting diode and the transmitted light amount detecting means to detect a rapid change in the transmitted light amount due to the setting of the sample cuvette. measuring device. 6. The measuring device according to claim 1, further comprising means for holding the sample cuvette at a constant temperature. 7. The measuring device according to claim 1, further comprising means for holding the temperature of the light emitting diode and / or the transmitted light amount detecting means at a constant temperature. 8. The measuring device according to claim 1, wherein the reaction reagent contains a horseshoe crab blood cell component. 9. The measuring device according to claim 1, wherein the reaction reagent contains a silkworm body fluid component. 10. The measuring device according to claim 1, wherein the reaction reagents are a horseshoe crab blood cell component or a silkworm body fluid component and a chromogenic synthetic substrate. 11. A method for measuring a microorganism-derived component based on the time until the amount of transmitted light changes by a predetermined ratio after a predetermined time has passed after the preparation of a mixture of a sample and a reaction reagent (test sample solution). , The test sample solution is stored in a sample cuvette and held in a stationary state, the test sample cuvette is irradiated with blue and / or purple light rays from a light emitting diode, and the amount of transmitted light of the irradiated light rays is detected. Then, a method for measuring a microorganism-derived component, which comprises detecting a time from the passage of the predetermined time until the amount of transmitted light of the test sample liquid changes by a predetermined ratio, and measuring the target substance based on the time. 12. The measuring method according to claim 11, wherein the change in the amount of transmitted light occurs in response to a gelling reaction, a melanin producing reaction or a dye releasing reaction occurring in the test sample solution.