JP2003329670A - Reagent for urinary sediment examination and method of urinary sediment examination - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To provide a reagent for urinary sediment examination that takes place of Steinheimer dyeing reagents and a new method of urinary sediment examination. <P>SOLUTION: An aqueous liquid containing Alcian green and pyronin dye is used as the reagent for urinary sediment examination. The reagent is added to urinary sediments at a ratio (volume ratio) of 1-50:1 (urinary sediments: reagent) and the urinary sediments are observed under a microscope. <P>COPYRIGHT: (C)2004,JPO

Description

Detailed Description of the Invention

[0001]

BACKGROUND OF THE INVENTION 1. Field of the Invention The present invention relates to a novel urine sediment inspection reagent and a urine sediment inspection method using the reagent.

[0002]

2. Description of the Related Art In medical examinations such as a physical checkup,
Urine sediment is examined as a primary screening for the detection of various diseases such as urinary tract diseases, nephritis, and cancer. More specifically, the precipitate (urine sediment) obtained by centrifuging fresh urine is stained with a staining reagent, and red blood cells, white blood cells, epithelial cells,
Observe the presence or absence of columnar crystal components. The staining method applied to this method was Sternheim, 1975.
Stenheimer Staining (A sup
ravital cytodiagnosticsta
in for urinary sediments,
J. A. M. A. , 231: 826-832, 197.
5) is the principle.

In this Stenheimer dyeing (hereinafter referred to as "S dyeing original method"), as a dye for dyeing the nucleus blue,
National First Blue, which is a phthalocyanine base dye, is used, and Pyronin B, which is a xanthene base dye, is used as a dye that stains the cytoplasm in reddish purple. However, in Japan, Alcian Blue, which is also a phthalocyanine base dye, has been used in place of National First Blue. This method unique to Japan (hereinafter referred to as "S staining modified method") is widely used as a method for staining urinary sediment in Japan. This modified S-staining method is superior to the original S-staining method in that it can also stain mucoprotein, which is the main component of the cast in urine.

However, last year, due to various reasons, Alcian Blue (Color Index No. 7424)
0, C.I. I. Ingrain Blue 1, as an example of a commercially available product, includes Alcian Blue 8GX manufactured by Attic Industries, Inc.) has been discontinued. Therefore, when the stock of Alcian blue has reached the bottom, more specifically, after a few years, the reagent for S-staining modified method containing Alcian blue will not be available.

[0005]

As described above, the modified S-staining method is an essential method for staining as a pretreatment for performing a microscopic examination of urinary sediment. Therefore, the search and provision of a staining reagent that uses a dye other than Alcian blue and can replace the reagent for the modified S staining method is eagerly desired in the field of clinical examination.

The present invention has been made in view of the above circumstances, and an object thereof is to provide a new urinary sediment inspection reagent used for dyeing urine sediment and a urine sediment inspection method using the reagent. To do.

[0007]

[Means for Solving the Problems] In order to solve the above problems,
The present inventor has conducted earnest research and, as a result, completed the present invention.

That is, the present invention provides a reagent for examining urinary sediment, which is an aqueous liquid containing Alcian Green and a pyronin dye.

As a pyronin dye, Color Index No. 45005 Pyronin and / or Color Index No. It is preferred to use 45010 of pyronin.

The above reagent preferably further contains an antiseptic and / or an antifungal agent.

As a solvent constituting the aqueous liquid, a buffer solution is preferable, a phosphate buffered saline and a Tris buffered saline are more preferable, and a phosphate buffered saline having a concentration of 10 times is particularly preferable.

In the present invention, the above-mentioned urinary sediment inspection reagent of the present invention is added to the urine sediment, and the urine sediment: reagent = 1 to 50: 1.
Provided is a method for examining urinary sediment, which comprises observing a part or all of the urinary sediment under a microscope in addition to the ratio (volume ratio).

It is preferable that the observation be performed within 1 hour after the reagent is added to the urine sediment.

It is preferable that the observation is performed at a magnification of 100 for the entire visual field.

It is preferable that the observation is carried out at a magnification of 400 times for 20 or more visual fields, if necessary.

[0016]

BEST MODE FOR CARRYING OUT THE INVENTION The present invention will be described in detail below.

The dyes used in the reagent of the present invention are Alcian Green and Pyronine dyes.

Alcian Green is a phthalocyanine dye. As an example of commercial products of Alcian Green, Imperial Chemical Indu
stories (ICI) "Alcian Green 3
BX (Color Index Ingrain Gr
een 1) "and"Industry's"Alcian Green 2GX (Color Inde
x IngrainGreen 2) ”.

Pyronine dye is a kind of red basic dye of xanthene (or xanthene) dye. Color
Index No. 45000 to 45020 dyes,
It is classified as a pyronin dye. In the present invention, Color
Index (color index) No. 45005
Pyronin and / or Color Index (Color Index) No. The use of 45010 of pyronin is preferred. C. I. No. As an example of a commercially available product of Pyronin of 450005, Pyronin G manufactured by Bayer Co. I. No. An example of a commercially available product of 45010 Pyronin is Bayer Pyronin B.

The reagent of the present invention is in the form of an aqueous liquid.
The solvent that constitutes this aqueous liquid is, for example, water (ion-exchanged water, distilled water) and various buffer solutions.

Examples of the buffer solution include phosphate buffered saline (PBS, around pH 7.4), PBS (-), 10x.
PBS (PBS having a reagent concentration of 10 times higher), Tris-hydrochloric acid buffer solution (pH about 7.2 to about 9.0), Tris-buffered saline (TBS, near pH 7.4), sodium phosphate buffer solution (about pH) 5.7 to about 8.0), citrate buffer (pH
About 3.0 to about 5.8), acetate buffer (pH about 3.6 to about 5.6), glycine-NaOH buffer (pH about 8.6 to
About 10.6), borax buffer (pH about 9.0 to about 10.
6, 40 ° C), boric acid / borax buffer (pH about 7.6-
About 9.2), carbonate / bicarbonate buffer (pH about 9.2 to about 1)
0.7), a buffer of Sdrensen and Palitrsch, and a buffer of Walpole (pH about 3.6 to about 5.
6), MeIlvaine buffer (pH about 2.2 to about 8.9), Amediol buffer (pH about 7.8 to about 9.9).
2) and barbital buffer (pH about 7.0 to about 8.
9) and the like.

The pH is 5.0 to 9.0, preferably 6.0.
It is preferable to use a buffer solution of about 8.0, more preferably about 7.0.

Among the buffer solutions, phosphate buffered saline (PBS) and Tris buffered saline (TBS) are preferable, and phosphate buffered saline (10 × PB) having a concentration 10 times that of phosphate buffered saline (10 × PB) is used.
Use of S) is particularly preferable because hemolysis of red blood cells is small.

The reagent of the present invention preferably further contains a preservative and / or a fungicide. By including these, it is possible to prevent the contamination of the reagent by bacteria or mold and the error of the test result derived therefrom.

Examples of preservatives used in the reagent of the present invention include sorbic acid, salicylic acid, dehydroacetic acid and N 2 thereof.
a salts, acids such as parahydroxybenzoic acid esters and undecylenic acid, phenol, cresol, p-chlorophenol, benzyl alcohol, p-chloro-m-xylenol, p-chloro-m-cresol, thymol,
Phenethyl alcohol, 0-phenylphenol, Irgasan (trademark), phenols such as hexachlorophen and chlorhexidine, alcohols such as ethanol and chlorobutanol, and benzalkonium chloride.
Examples thereof include cationic surfactants such as benzethonium chloride, cetylpyridinium chloride, cetyltrimethylammonium bromide, and myristyldimethylbenzylammonium chloride.

Further, examples of the fungicide used in the reagent of the present invention include benzalkonium chloride, benzethonium chloride,
Examples include cationic surfactants such as cetylpyridinium chloride, cetyltrimethylammonium bromide, myristyldimethylbenzylammonium chloride.

The reagent (aqueous liquid) of the present invention contains Alcian green, for example, 0.2 to 10% by weight, generally 0.3 to 5% by weight, preferably 0.5 to 2.5% by weight. ,
More preferably 1.0-1.6 wt%, even more preferably 1.1-1.5 wt%, most preferably 1.2-
Contains 1.4% by weight. The reagent of the present invention contains a pyronin dye, for example, 0.1 to 5% by weight, generally 0.15 to 3%.
% By weight, preferably 0.2 to 1.5% by weight, more preferably 0.2 to 0.8% by weight, even more preferably 0.
3 to 0.7% by weight, most preferably 0.4 to 0.6% by weight. The reagent of the present invention has a color index N.
o. 45005 Pyronin (Pyronin G) and Color Index No. 45010 Pyronine (Pyronin B)
When both are contained, the reagent of the present invention (aqueous liquid)
Each of these pyronins, for example, 0.05-2.
5% by weight, generally 0.075 to 1.5% by weight, preferably 0.1 to 0.75% by weight, more preferably 0.1.
.About.0.4 wt%, and even more preferably 0.15 to 0.
35% by weight, most preferably 0.2 to 0.3% by weight. Color index No. 45005 Pyronin (Pyronin G) and Color Index No. 450
When both of 10 pyronins (pyronin B) are used, the staining property of the cytoplasm is particularly excellent.

The content of the antiseptic agent and the antifungal agent depends on the kind of the compound (antiseptic agent, antifungal agent) used. The proper content of each antiseptic agent and antifungal agent is known in the technical field, but as an example, in the case of benzethonium chloride, it is generally 0.0
2 to 0.08% by weight, preferably 0.03 to 0.07% by weight, more preferably 0.04 to 0.06% by weight, and most preferably around 0.05% by weight. Further, in the case of sodium azide, the best value is around 0.1% by weight.

The reagent of the present invention comprises an aqueous liquid, dyes, and
It can be produced by dissolving an antiseptic and a fungicide, if necessary. However, prepare a stock solution of each dye (one kind of dye dissolved in a solvent), mix them at an appropriate ratio, and if necessary, dissolve the preservative and antifungal agent in the resulting mixture. It is preferable to manufacture by the method.

In the present invention, the above-mentioned urine sediment inspection reagent of the present invention is added to the urine sediment, and the urine sediment: reagent (volume ratio) = 1 to 50:
1, preferably 3 to 30: 1, more preferably 5 to 2
A method for examining urinary sediment, which comprises observing a part or all of the urinary sediment under a microscope by adding in a ratio of 0: 1, even more preferably 8 to 18: 1, most preferably 10 to 16: 1. provide.

Observation of the stained urine sediment by a microscope is as follows.
It is performed after the cytoplasm and nucleus of the components contained in the urine sediment are stained. Since the reagent of the present invention has good dyeability and is hard to be deeply dyed, the observation can be performed from immediately after adding the reagent to the urine sediment to about 2 hours later. In addition,
The observation is preferably performed within 1 hour after adding the reagent to the urine sediment.

The observation was carried out at a microscope magnification of 100-40.
It is preferable to set it to 0 times, 100 times or 400 times
It is particularly preferable to carry out at a double magnification. For example, when observing at a magnification of 100 times, it is preferable to observe the entire visual field, and when observing at a magnification of 400 times, for example, it is preferable to observe 20 or more visual fields. At a magnification of 100 times, it is usually determined whether or not the prepared urine sediment sample is not biased or does not contain a mass of sediment. At a magnification of 400 times, the presence or absence of various components is determined and the number is calculated.

Conventionally, unstained urine sediment specimens and specimens stained by the S-staining modified method depending on the situation of the specimens were observed under a microscope to determine the number of red blood cells in one visual field,
The number of white blood cells, the amount of epithelial cells, the number of atypical epithelial cells, the number of casts, the amount of crystal components, etc. were determined. In the inspection method of the present invention, the stained urine sediment sample is observed, but the observation of the unstained sample may or may not be performed depending on the condition of the sample.

[0034]

EXAMPLES Hereinafter, the present invention will be described more specifically by showing examples.

(Example 1) (1) 10 ml of urine whose urinary sediment was confirmed to contain solid components such as red blood cells, white blood cells, and casts was placed in a centrifuge tube at room temperature at 500 g for 5 minutes. It was centrifuged. Each of the urine sediments thus obtained was stained with the dyes shown in Table 1 and examined microscopically to compare the staining properties. The stain is
Blue-based Alcian Green 3BX, Victoria Blue B, Toluidine Blue N, Cresyl Violet, Anilin Blue Water-soluble, Water Blue. Water Blue) and Astra Blue (Astra
An aqueous solution of 1.33% by weight for each dye of blue) and 0.5% by weight for each of red (Orange (orange) G, Pyronine (pyronine) G and Pyronine (pyronine) B dyes was prepared. Urine sediment 20
15 μl of an aqueous dye solution was used per 0 μl, and 5 minutes after the aqueous dye solution was added to the urinary sediment, observation was performed under a microscope at a magnification of 400 times. The results are shown in Table 1.

[0036]

[Table 1]

From the above results, Alcian Green 3
BX, Astra Blue, Pyronin G and Pyronin B
However, it was considered that it can be used as a dye for a reagent for staining urinary sediment.

(2) Next, Alcian Green 3BX
The urinary sediment was stained with an aqueous solution of a mixture of aspirin and pyronin G and an aqueous solution of a mixture of astra blue and pyronin G. As is clear from Table 1, the results for Pyronin B were almost the same as those for Pyronin G, so no test was performed here. Staining solution is Alcian Green 3B
An aqueous solution containing 1.33% by weight of X or Astra Blue and 0.5% by weight of Pyronin G was prepared. Staining and observation
It carried out on the same conditions as what was described in (1). The results are shown in Table 2.

[0039]

[Table 2]

From the above results, Alcian Green 3BX, Pyronin G and Pyronin B were selected as candidates for the dye used in the reagent for staining urinary sediment.

(Example 2) In order to examine the optimum combination of dyes used in the reagent for staining urinary sediment, Alcian Green 3BX, Pyronin G, Alcian Green 3B.
X and Pyronin B and Alcian Green 3BX, Pyronin G and Pyronin B, and benzethonium chloride as a preservative (Wako Pure Chemical Industries, Ltd., Lot.DW
L-5057, powder, purity 98% or more) was also prepared. More specifically, first, a 2 wt% aqueous solution of Alcian Green 3BX, a 1.5 wt% aqueous solution of pyronin G, a 1.5 wt% aqueous solution of pyronin B and a 10 wt% aqueous solution of benzethonium chloride were prepared, and These aqueous solutions were mixed in the proportions described in 3.

[0042]

[Table 3]

Using these three types of aqueous dye solutions,
Urine sediment was stained. Staining and observation are done in Example 1
It carried out on the same conditions as what was described in (1). The results are shown in Table 4.

[0044]

[Table 4]

As shown in Table 4, any of aqueous dye solutions can be used as a reagent for staining urinary sediment.
The aqueous dye solution was excellent in terms of stainability of cells and casts and cleanliness of the background.

Next, a dye aqueous solution-b was prepared in the same manner as the dye aqueous solution except that 10 × PBS was used instead of water. The dye solution-b was used to similarly stain and observe the urine sediment. As a result, the dyeability of the urinary sediment component including the dyeability of the cytoplasm and the nucleus and the dyeability of the column and the cleanliness of the background were more excellent than those using the aqueous dye solution.

(Example 3) (1) Urine (specimens 1 to 3) was prepared in which the urine sediment was confirmed to contain solid components such as red blood cells, white blood cells and casts. The pH of the sample 1 was about 5.0. The pH of the sample 2 is about 7.0
Met. The pH of the sample 3 was about 9.0. Sample 1
When the specific gravity of each of 3 was measured using a densitometer (by a refraction method), all were in the range of 1.015 to 1.020.

Specimen 1 was placed in a centrifuge tube by 10 ml and centrifuged at 500 g for 5 minutes at room temperature.
Each of the urinary sediments (200 μl) derived from the specimen 1 thus obtained was treated with one of the staining solutions shown in Table 5.
5 μl was added.

[0049]

[Table 5]

Five minutes after the addition of the staining solution to the urine sediment, the number of remaining red blood cells that did not undergo hemolysis was counted under a microscope at a magnification of 200 times or 400 times. More specifically, Neub was used to calculate the number of red blood cells in 1 μl of urine sediment containing staining solution.
The number of red blood cells was counted for each of the 5 large sections using an Auer counting board, and the average value thereof was calculated.

With respect to the samples 2 and 3, the same processing and observation as in the sample 1 were performed.

The results are shown in FIGS. 1 and 2. FIG. 1 is a bar graph showing the number of remaining red blood cells that were not hemolyzed.
[Fig. 4] is a bar graph showing the results of expressing the residual red blood cell count of each sample as a percentage with respect to the residual red blood cell count in the case of no staining.

As is apparent from these figures, Invention Example a
Is used for acidic urine (pH about 5.0), which is equivalent to the case of using the commercially available reagent for modified Stenheimer staining, and for neutral urine (pH about 7.0), modified for commercially available Stenheimer staining. Less than with reagents, and alkaline urine (p
H about 9.0), hemolysis was observed to be slightly higher than that when a commercially available reagent for modified Stenheimer staining was used. Comprehensively judging from these results, it can be said that the invention example a shows the performance equal to or higher than that of the commercially available reagent for modified Stenheimer staining. Especially, when the urine is neutral, the number of red blood cells was equivalent to that of non-staining even when stained with the invention example a, so that it is possible to eliminate the need for observation without staining in a sample having a neutral pH. There is a nature.

(2) Urine (pH about 7.0) 3 in which the urine sediment was confirmed to contain solid components such as red blood cells, white blood cells and casts.
Species (specimens 4-6) were prepared. When the specific gravities of these samples were measured using a pycnometer (by refractometry), sample 4 was 1.005, sample 5 was 1.020, and sample 6 was 1.
It was 030.

Thereafter, the number of remaining red blood cells in the urine sediment was counted by the same method as described in (1).

The results are shown in FIGS. 3 and 4. FIG. 3 is a bar graph showing the number of remaining red blood cells that were not hemolyzed.
[Fig. 4] is a bar graph showing the results of expressing the residual red blood cell count of each sample as a percentage with respect to the residual red blood cell count in the case of no staining.

As is clear from these figures, Invention Example a
With low specific gravity urine (specific gravity 1.005), it is considerably less than when using the commercially available reagent for modified Stenheimer's stain. For medium specific gravity urine (specific gravity 1.020), modified for commercially available Stenheimer's stain. A little more than when using reagents,
In high specific gravity urine (specific gravity 1.030), hemolysis was observed, which was equal to or slightly less than the case of using a commercially available reagent for modified Stenheimer staining. Comprehensively judging from these results, it can be said that the invention example a shows the performance equal to or higher than that of the commercially available reagent for modified Stenheimer staining. Particularly, in the case of low specific gravity urine, the number of erythrocytes was the same as that in the case of staining without using the invention example a, so that observation without staining may be unnecessary.

Example 4 (1) An experiment similar to that of Example 3 (1) was conducted, except that the invention example b shown below was used instead of the invention example a.

Invention Example b: [Alcian Green 3BX
(2 wt%) 10 × PBS solution 2.0 ml] + [pyronin G (1.5 wt%) 10 × PBS solution 0.5 m
l] + [0.5 ml of 10 × PBS solution of pyronin B (1.5% by weight)] + [benzethonium chloride (10% by weight)
Aqueous solution of 30 μl]

The results are shown in FIG. FIG. 5 is a bar graph showing the results of expressing the number of remaining red blood cells of each sample as a percentage with respect to the number of red blood cells remaining without hemolysis in the case of no staining.

As is apparent from FIG. 5, when the invention example b is used, acidic urine (pH about 5.0) and neutral urine (pH about 7.
In 0), hemolysis was less than in the case of using the existing reagent for modified Stenheimer's staining and not in the case of non-staining. Also, in the case of alkaline urine (pH about 9.0), hemolysis was equivalent to that using the existing reagent for modified Stenheimer staining. Comprehensively judging from these results, it can be said that the invention example b is lower in hemolytic property and superior to the existing reagents for modified Stenheimer staining such as commercial products. Especially when the urine was acidic to neutral, the one stained with Inventive Example b had more red blood cells than the unstained sample. Therefore, the sample with acidic to neutral pH was observed without staining. May be unnecessary.

(2) The same experiment as in Example 4 (2) was carried out except that Invention Example b was used instead of Invention Example a.

The results are shown in FIG. FIG. 6 is a bar graph showing the results of expressing the number of residual red blood cells of each sample as a percentage with respect to the number of red blood cells remaining without hemolysis in the case of no staining.

As is apparent from FIG. 6, when the invention sample b is used, low specific gravity urine (specific gravity 1.005) is equivalent to or less than the case of using the existing reagent for modified Stenheimer staining. There is less specific gravity urine (specific gravity 1.020) than when using the existing reagent for modified Stenheimer staining.
In high specific gravity urine (specific gravity 1.030), hemolysis was observed, which was almost the same as when a commercially available reagent for modified Stenheimer staining was used. Therefore, it can be said that Invention Example b has lower hemolytic activity and is more excellent than existing reagents for modified Stenheimer staining, including commercially available products. In addition, when stained using Invention Example b, urine of any specific gravity was 70% of that in the case of no staining.
Since the number of remaining red blood cells is shown above, there is a possibility that observation without staining may be unnecessary.

[0065]

The alcian green used for the production of the reagent of the present invention is more soluble in an aqueous liquid than the alcian blue used in the modified S dyeing method.
Therefore, the reagent for staining the urinary sediment is easily manufactured, and the energy consumed for manufacturing the reagent is greatly saved.

When the reagent of the present invention is used, the subject is less likely to be deeply stained than in the case where the existing reagent for modified S-staining method is used. Therefore, it becomes easier to detect the dyed component. Further, for the same reason, the contrast between the stained portion and the non-stained portion is large and the state lasts longer, so that it is less necessary to observe both the unstained specimen and the stained specimen. That is, the examination may be only a microscopic examination of the stained specimen, and the examination efficiency is improved.

In the urinary sediment examination method of the present invention, the existing color atlas can be used as it is. Therefore, a person involved in the clinical examination does not need to learn a new technique for examining the urine sediment.

In addition to Alcian Green, Color Index No. 45005 Pyronine (Pyronin G)
And color index No. When the reagent of the present invention containing both 45010 of pyronin (pyronin B) is used, the dyeing property of the nucleus is particularly excellent, and thus the observation of the intranuclear structure becomes easier.

When the reagent of the present invention using a PBS solution having a concentration of about 10 times is used as a solvent, hemolysis of erythrocytes is less likely to occur, and dyeability is particularly excellent.

[Brief description of drawings]

FIG. 1 shows urine of three pH values (specific gravity: 1.01).
5 to 1.020) for each urinary sediment, a bar graph showing the number of remaining red blood cells after staining with the existing reagent for modified Stenheimer staining and the reagent of the present invention (aqueous medium: water) Is.

FIG. 2 is a bar graph showing the data of FIG. 1 with the residual red blood cell count in the case of no staining being 100%.

FIG. 3 shows the existing reagent for modified Stenheimer staining and the reagent of the present invention (aqueous medium: water) for each urine sediment obtained from urine of three types of specific gravity (pH about 7.0). 3 is a bar graph showing the number of remaining red blood cells after being stained with.

FIG. 4 is a bar graph showing the data in FIG. 3 with the residual red blood cell count in the case of no staining being 100%. It is a graph.

FIG. 5 shows urine with three pH values (specific gravity: 1.01).
5 to 1.020), the residual erythrocyte count after staining with the existing reagent for modified Stenheimer staining and the reagent of the present invention (aqueous medium: 10 × PBS) 2 is a bar graph showing the number of remaining red blood cells in the case of no staining as 100%.

[Fig. 6] Fig. 6 shows the existing reagent for modified Stenheimer staining and the reagent of the present invention (aqueous medium: 10x) for each urine sediment obtained from urine of three types of specific gravity (pH about 7.0). PB
It is a bar graph showing the number of residual red blood cells after staining with S), with the number of residual red blood cells in the case of no staining being 100%.

Claims (10)

[Claims] 1. A reagent for examining urinary sediment, which is an aqueous liquid containing Alcian Green and a pyronin dye. 2. The color index No. 1 is a pyronine dye. 45005 Pyronin and / or Color Index No. The reagent for urinary sediment examination according to claim 1, which is 45010 of pyronin. 3. A preservative and / or a fungicide are further included,
The reagent for urinary sediment examination according to claim 1 or 2. 4. The urinary sediment examination reagent according to claim 1, wherein the solvent forming the aqueous liquid is a buffer solution. 5. The reagent for urinary sediment examination according to claim 4, wherein the buffer solution is phosphate buffered saline or Tris buffered saline. 6. The reagent for urinary sediment examination according to claim 4, wherein the buffer solution is a 10-fold concentrated phosphate buffered saline. 7. The urine sediment according to any one of claims 1 to 6.
A method for inspecting urinary sediment, which comprises observing a part or all of the urinary sediment under a microscope by adding the reagent described in the item 1 in a ratio of urine sediment: reagent = 1 to 50: 1 (volume ratio). 8. The method for examining urinary sediment according to claim 7, wherein the observation is performed within 1 hour after adding a reagent to the urine sediment. 9. The method for examining urinary sediment according to claim 7, wherein the observation is performed at a magnification of 100 for the entire visual field. 10. The method for examining urinary sediment according to claim 7, wherein the observation is carried out at a magnification of 400 times for 20 or more visual fields as needed.