KR101997193B1 - A method for smear treatment of specimen - Google Patents

Abstract

A sample smear processing method is provided. The smear treatment method comprises preparing a specimen having a bacterium whose infectivity has been removed in the state that the cell wall is not destroyed, and a step of applying the specimen to a glass.

Description

A METHOD FOR SMEAR TREATMENT OF SPECIMEN [0002]

More particularly, the present invention relates to a method for smearing a specimen for detection of Mycobacterium tuberculosis .

Tuberculosis is an infectious disease characterized by latent, progressive, and active infection with Mycobacterium , the causative organism. Specifically, Mycobacterium tuberculosis , Mycobacterium bovis , Mycobacterium avium , and other non-human mammals and birds may be infected with M. tuberculosis. However, human infection is mainly caused by Mycobacterium tuberculosis Mycobacterium tuberculosis is the major causative organism. Mycobacterium tuberculosis has a rod shape with a thickness of 0.2 to 0.5 μm and a length of 1 to 4 μm. Mycobacterium tuberculosis is an antioxidant bacterium that can survive for a long time under dry conditions and is resistant to strong acids and alkalis because it is surrounded by cell walls with many fat components.

Tuberculosis spreads mainly from person to person through air. When a contagious tuberculosis patient speaks, coughs or sneezes, it is dispersed in the air in droplets containing Mycobacterium tuberculosis. At this time, the scattered tubercle bacillus invades into the lungs of the surrounding people and propagates and the infection is made.

Tests that are commonly used to diagnose tuberculosis include skin reaction test, staining and culturing test, and chest x-ray test through smear test.

Korean Patent No. 10-1129806

Among the tests used for the diagnosis of tuberculosis, the method is a method in which specimens such as human sputum are permeated through a filter to remove impurities and then thinly coated on slides to selectively stain only mycobacteria and to observe them with a microscope. Collecting hammer test is a simple staining method for bacteriological diagnosis. Since it is relatively simple and quick to detect the infection as compared with other methods, it is possible to quickly detect and treat patients with active TB disease.

However, since the collecting hammer test is carried out by collecting a sample from a liquid specimen suspected of having a tuberculosis bacterium, an examiner, such as a clinician or a pathologist, is always exposed to the tuberculosis bacterium. For example, in the process of moving a sample provided from a patient, filtering a sample to collect a sample, smearing a sample, or observing a microscope of a slide on which a sample is smear, the tubercle bacilli are spread into the air, There is a risk of infection.

Therefore, it is required to develop a new test method which is comparatively simple and quick, and to prevent the infected tuberculosis infection of the examinee.

Accordingly, a problem to be solved by the present invention is to provide a new smear treatment method capable of inhibiting the infection of M. tuberculosis by an inspector.

The present invention has been made in view of the above problems, and it is an object of the present invention to provide a method of manufacturing the same.

According to an aspect of the present invention, there is provided a method for smear treatment of a specimen, comprising the steps of: preparing a specimen having infectivity-free microorganisms in the absence of destruction of a cell wall; .

The step of preparing the specimen having the infectious microorganism bacterium comprises preparing a specimen container containing a specimen having Mycobacterium mixed with a bactericidal reagent and containing the specimen having the infectious bacteria removed therein, The bactericidal reagent may include sodium hydroxide, sodium hypochlorite, sodium citrate, and a solvent.

The step of preparing a sample container containing the specimen mixed with the specimen and the bactericide reagent may include a step of receiving the specimen having the first internal space sealed with the bacteria killing reagent accommodated therein and the bacteria having no infectivity removed, Preparing a sample container including a first inner space and a second inner space configured to be openable and closable, separated from the first inner space, and supplying the germicidal reagent accommodated in the first inner space of the sample container to the second inner space Contacting the specimen with the bactericidal reagent to the specimen having the infectious bacteria not removed from the infectious agent, and mixing the specimen with the anticancer reagent in the second internal space of the specimen container to remove the infectious agent And forming a specimen having bacteria.

The sample container may further include a cap portion having the first inner space and a sample container portion coupled to the cap portion to form the second inner space, wherein the cap portion includes a coupling portion A pressing portion disposed on an upper side of an inner circumferential surface of the annular fastening body; a side wall portion protruding downward from an edge of the pressing portion; a protrusion protruding downward from a center of the pressing portion to be inserted and disposed in the side wall portion; And a sealing sheet disposed at the lower end of the side wall portion and forming the first inner space together with the pressing portion and the side wall portion.

Wherein the step of contacting the specimen with the germicidal reagent comprises the step of applying a downward pressure to the pressing portion so that the cut portion protruding downward from the pressing portion partially cuts the sealing sheet, And injecting a reagent into the second internal space of the sample container.

The step of preparing a sample container containing a first inner space containing the bactericidal reagent and a second inner space containing the sample may include disposing a specimen having the infectivity- And a second internal space sealed by the coupling of the cap portion and the specimen accommodating portion may be formed.

In addition, the step of contacting the specimen with the bactericide reagent and the step of mixing the specimen with the bactericide reagent to form a specimen having the infectious microorganism, wherein the cap portion of the specimen container and the specimen accommodating portion And the specimen having the infectious bacterium and the specimen having the infectious bacterium can maintain the state of being shielded from the outside air.

A step of mixing the specimen with the anticancer reagent to form a specimen having the infectious bacteria and a step of grinding the specimen to the glass, The method comprising the steps of:

The step of applying the specimen to the glass may include a step of laminating the specimen having the infectious agent removed to the glass using an automatic smashing apparatus.

In addition, the step of applying the specimen having the infectious bacteria to the glass using the automatic smearing apparatus may include a smear processing container body having an internal space, a filter membrane inserted and disposed in the smear processing container body, The method comprising the steps of: preparing a smear processing container including a smear membrane inserted in a treatment vessel body and disposed below the filter membrane; a sample having infectious bacteria contained in the second inner space of the sample container, Injecting the sample into the internal space of the container, inserting the smear processing container containing the specimen having the infectious bacteria removed therein into the automatic smearing apparatus, sucking the specimen into the smear membrane side, and using the filter membrane Filtering said specimen having said infectious organisms removed, Inserting the glass between the membrane and the smear membrane, the method comprising the steps of: inserting a glass having a first face opposing the filter membrane and a second face opposing the smear membrane; And smearing the specimen on the second side of the glass by contacting the smear membrane.

The details of other embodiments are included in the detailed description.

According to the method for smear treatment of a specimen according to an embodiment of the present invention, it is possible to prevent the infection of Mycobacterium tuberculosis of an examiner who performs smear test by smearing the specimen after selectively removing the infectivity of the bacteria.

The effects according to the embodiments of the present invention are not limited by the contents exemplified above, and more various effects are included in the specification.

1 is a flowchart showing a smear processing method according to an embodiment of the present invention.
2 is a perspective view of a sample container used in a step of pretreating the sample of FIG.
3 is an exploded perspective view of the sample container of Fig.
4 is a cross-sectional perspective view of the sample container of Fig.
Fig. 5 is a sectional view of the sample container of Fig. 2; Fig.
6 is a perspective view of the slit portion of Fig.
Fig. 7 is a view showing the step of receiving the sample of Fig. 1;
Fig. 8 is a view showing a step of sealing the sample container of Fig. 1;
9 is a view showing the step of contacting the specimen with the bactericidal reagent of FIG.
FIG. 10 is a view showing a step of mixing the bactericidal reagent of FIG. 1 and a specimen.
11 is a view showing a step of opening the sample container of Fig.
12 is a perspective view of the smear processing container used in the step of smearing the pretreated sample of Fig. 1 into a glass.
13 is an exploded perspective view of the smear processing container of Fig.
14 is a cross-sectional view of the smear processing container of Fig.
15 is a view showing a step of injecting a specimen into the smear processing container of Fig.
16 is a view showing the step of inserting the smear processing vessel of Fig. 1 into the automatic smelting apparatus.
Fig. 17 is a view showing a step of filtering the specimen of Fig. 1; Fig.
Fig. 18 is a view showing a step of inserting the glass of Fig. 1;
19 is a view showing a step of applying a specimen to the glass of Fig. 1;
20 is a view showing a glass in a state in which bacteria are smoothed.

BRIEF DESCRIPTION OF THE DRAWINGS The advantages and features of the present invention and the manner of achieving them will become apparent with reference to the embodiments described in detail below with reference to the accompanying drawings. The present invention will now be described more fully hereinafter with reference to the accompanying drawings, in which preferred embodiments of the invention are shown. However, it will be understood by those of ordinary skill in the art that various changes in form and details may be made therein without departing from the spirit and scope of the invention. To fully disclose the scope of the invention to a person skilled in the art, and the invention is only defined by the scope of the claims.

The term " tuberculosis " as used herein means an infectious disease caused by infection with a Mycobacterium complex including tubercle bacillus, bovine tubercle bacillus, and moldy tubercle bacillus.

As used herein, the term " to " means a numerical range including a lower limit and an upper limit of the numerical value described before and the numerical value described later.

Hereinafter, the smear processing method of a specimen according to the present invention will be described in detail.

1 is a flowchart showing a sample smear processing method according to an embodiment of the present invention.

Referring to FIG. 1, the smear processing method according to the present embodiment includes a step (S100) of pretreating a specimen and a step (S300) of applying a pretreated specimen to a glass.

The step of pre-treating the specimen (S100) may be a step of preparing specimens having infectious bacteria. For example, the bacterium may be in a state in which only the infectious agent is selectively removed in the state where the cell wall is not destroyed.

In an exemplary embodiment, the step (S100) of pre-treating a specimen comprises preparing a specimen container containing a specimen having a bactericidal reagent and a specimen having no infectivity removed (S110), adding a bactericide reagent and a non- (S120), and a step (S130) of forming a specimen having infectious bacteria by mixing the specimen with the bactericide reagent and the specimen having no infectivity removed (S130).

First, a step (S110) of preparing a specimen container containing a specimen having a bacteria killing reagent and a specimen having no infectivity is described in detail.

(S110) of preparing a specimen container containing a specimen having a bactericidal reagent and a specimen having no infectivity removed, the specimen having the first internal space in which the anticancer reagent is contained and sealed, And a second inner space accommodated and separated from the first inner space and configured to be openable and closable. The step (S110) of preparing a sample container containing a specimen having a bactericidal reagent and a bacterium not having infectivity removed includes a step (S111) of preparing a specimen container including a first inner space and a second inner space.

2 is a perspective view showing the sample container 100 used in the smear processing method according to the present embodiment. 3 is an exploded perspective view of the sample container 100 of FIG. 4 is a cross-sectional perspective view of the sample container 100 of FIG. 5 is a sectional view of the sample container 100 of FIG.

1 to 5, the sample container 100 according to the present embodiment includes a cylindrical sample receiving portion 130 coupled with the cap portion 110 and the cap portion 110, (S1) and a second internal space (S2) configured to be openable and closable. The sample container 100 may be a container used for storing the sample and for destroying the bacteria contained in the sample. For example, the fungus may be homogeneous belonging to the Mycobacterium group.

The cap portion 110 may have a first internal space S1 in which a bactericidal reagent CP can be received. Specifically, the cap part 110 includes an annular fastening body 111, a pressing part 112 disposed on the upper side of the inner circumferential surface of the annular fastening body 111, a side wall part 112 protruding downward from the edge of the pressing part 112, A cutout portion 114 protruding downward from the center of the pressing portion 112 and a sealing sheet 115 disposed at a lower end of the side wall portion 113. [ The cap part 110 may be at least partly placed in the sample accommodating part 130 in a state where the cap part 110 and the sample receiving part 130 are fastened.

The fastening body 111 may be a portion that abuts and is coupled to the sample accommodating portion 130. For example, the annular fastening body 111 may have a reversible engaging means capable of engaging with the sample receiving portion 130. As used herein, the term " reversible engagement means " means engagement means capable of repeatedly engaging and disengaging.

In the exemplary embodiment, the annular fastening body 111 may have a first thread 111a formed on the lower side of the inner circumferential surface and may be screwed with the second thread 130a of the upper outer circumferential surface of the sample accommodating portion 130 . However, the present invention is not limited thereto, and in another embodiment, the fastening body 111 and the sample receiving part 130 may be clip-coupled or slide-coupled.

The pressing portion 112 may be formed in a circular shape on the upper side of the inner peripheral surface of the fastening body 111 to form a cover constituting the upper end of the cap portion 110. Further, the pressing portion 112 can be configured to deform the shape when an external force is applied. In an exemplary embodiment, the pusher 112 may include an elastic dome that is approximately centered in a plane. The elastic dome may have an upward convex shape so that its center is higher than the edge. When the downward pressure is applied to the elastic dome, the pushing part 112 can be moved downward as a whole. In some embodiments, when the external force is removed, the resilient dome can be restored to its original shape by elastic force. In this case, the pressing portion 112 may further include a restoring rib to increase the restoring force of the elastic dome.

The side wall portion 113 protrudes downward from the edge of the pressing portion 112 to form a side wall of the cap portion 110. The side wall portion 113 can be inserted and disposed in the sample accommodating portion 130. The upper end of the side wall portion 113 may be connected to the pressing portion 112. The cross section of the side wall section 113 may have an annular shape and the side wall section 113 may have a substantially cylindrical shape with a lower end opened. In this case, the outer diameter of the side wall portion 113 may be smaller than the inner diameter of the fastening body 111. That is, the outer wall of the side wall portion 113 may be spaced inward from the inner peripheral surface of the annular fastening body 111.

The cut-out portion 114 may protrude downward from the center of the pressing portion 112 and may be inserted and disposed in the side wall portion 113. The upper end of the cutout 114 may be connected to the pushing part 112 and the lower end of the cutout 114 may be located above the sealing sheet 115. Although the lower end of the cut-out portion 114 and the upper surface of the sealing sheet 115 are illustrated as being separated from each other in the drawing, the lower end of the cut-out portion 114 may be in contact with the upper surface of the sealing sheet 115 have. When the pressing part 112 moves downward due to external force, the cut part 114 connected to the pressing part 112 can move downward together with the pressing part 112. [ Whereby the cut-out portion 114 can cut out the approximately central portion of the sealing sheet 115. [

The sealing sheet 115 may be disposed at the lower end of the side wall part 113 to form a cover which forms the lower end of the cap part 110. The sealing sheet 115 may be made of a material which does not transmit the liquid composition. That is, the sealing sheet 115 may be made of a material that can prevent permeation of the bactericidal reagent CP in the first internal space S1. For example, the sealing sheet 115 may be formed of a silver foil or a synthetic resin thin film material. The sealing sheet 115 can form the first internal space S1 of the cap portion 110 together with the pressing portion 112 and the side wall portion 113. [

In the first internal space S1, a bactericidal reagent CP can be received and sealed. For example, the bactericidal reagent (CP) may be a composition that selectively kills bacteria belonging to the Mycobacterium group.

The bactericidal agent (CP) according to this embodiment includes sodium hydroxide (NaOH), sodium hypochlorite (NaOCl), sodium citrate, and a solvent.

The sodium hydroxide acts as an emulsifier and a decontamination agent so that when the specimen contains a mucus substance such as sputum, it can decompose the mucus material and inhibit the growth of bacteria. In addition, the viscosity of a specimen can be lowered by decomposing a mucus substance (for example, mucin) to facilitate smearing of a specimen on a slide, thereby obtaining a clearer image. Further, as the viscosity of the specimen is lowered, the filtering characteristics of the specimen can be improved. On the other hand, if the content of sodium hydroxide is too small, the ability to decompose the mucus material may be markedly deteriorated.

The sodium hypochlorite may have a selective killing or bactericidal action against Mycobacterium. For example, hypochlorous acid (HClO), which is generated when sodium hypochlorite is diluted in a solvent (e.g., water), permeates the cell membrane of Mycobacterium to remove enzymatic action, and when sodium hypochlorite is diluted in a solvent Oxygen generated can oxidize the cell membrane to maximize the induction of death.

In an exemplary embodiment, the sodium hypochlorite content may be greater than the sodium hydroxide content. For example, the upper limit of the sodium hypochlorite content may be about 180 parts by weight, about 170 parts by weight, or about 150 parts by weight based on 100 parts by weight of sodium hydroxide. The lower limit of the content of sodium hypochlorite may be about 110 parts by weight based on 100 parts by weight of sodium hydroxide. If the sodium hypochlorite is contained in an amount of less than 110 parts by weight, the killing characteristic for the Mycobacterium may be insufficient. In addition, dyeing and smear inspection of a sample may affect dyeing characteristics. If the sodium hypochlorite is contained in an amount of more than 180 parts by weight, precipitates may be formed during long-term storage of the composition, which may hinder storage stability. In addition, the microorganism other than Mycobacterium in the specimen or the background cells may be destroyed and the dyeing characteristics of the smear sample may be deteriorated.

Said citrate can chelate heavy metal ions in a specimen such as a sputum, thereby promoting the function of the sputum lytic enzyme and contributing to stabilization.

In an exemplary embodiment, the content of sodium citrate may be less than the content of sodium hydroxide. For example, the upper limit of the content of sodium citrate may be about 90 parts by weight, or about 80 parts by weight, based on 100 parts by weight of sodium hydroxide. The lower limit of the content of sodium citrate may be about 60 parts by weight or about 65 parts by weight based on 100 parts by weight of sodium hydroxide. If the amount of sodium citrate is less than 60 parts by weight, complete chelation between heavy metal ions in the sample may be difficult. In addition, when the amount of the citric acid is more than 90 parts by weight, the sodium hypochlorite death property can be lowered.

The solvent is not particularly limited as long as it can dissolve sodium hydroxide, sodium hypochlorite and sodium citrate, but may be, for example, distilled water or deionized water.

As a non-limiting example, the bactericidal reagent CP comprises about 3.5% to 4.0% by weight of sodium hydroxide, about 4.5% to 6.0% by weight of sodium hypochlorite, 2.4% to 3.0% And a remaining amount of the solvent. For example, the solvent may be distilled water and the composition may comprise only sodium hydroxide, sodium hypochlorite, sodium citrate, and 87.0 wt% to 89.6 wt% of the distilled water.

The bactericidal reagent (CP) according to the present embodiment can selectively kill only microorganisms belonging to the Mycobacterium group. For example, when contacted with microbacterium in a human sputum, the microbicide (CP) selectively kills microbacterium and does not kill other microorganisms, but the present invention is not limited thereto. As used herein, the term 'death' refers to the elimination of the infectivity of the subject organism, while keeping the peculiar shape of the organism to be killed in a visually recognizable manner.

In addition, when the specimen has a viscosity including a mucus substance such as human sputum, there is a problem that it is difficult to filter the membrane or to apply the sputum thinly to the slide in a filtering step (S340) to be described later. In addition, since the sputum has a viscosity, there is a problem that the smear surface is uneven and rugged. However, the bactericidal reagent (CP) according to the present embodiment is mixed with sputum to decompose the mucin in the sputum, thereby lowering the viscosity, thereby improving the workability. When the sputum is applied to the slide, There is an effect that can be.

In addition, the bacteriostatic agent (CP), which may be in contact with the human body, can be kept sealed in the first internal space S1. Therefore, the bactericidal reagent (CP) can not only be free from external contamination but also can prevent harmful effects to the patient or the inspector.

The sample storage part 130 may be a cylindrical container with an open upper part. The sample receiver 130 can provide a space for storing specimens such as human sputum. The sample receiving portion 130 may have a reversible coupling means capable of being coupled with the cap portion 110, specifically, the coupling body 111 of the cap portion 110. In the exemplary embodiment, the specimen accommodating portion 130 can be screwed to the first screw thread 111a formed on the lower side of the inner circumferential surface of the fastening body 111 with the second screw thread 130a formed on the upper outer circumferential surface. In another embodiment, the sample receiving portion 130 and the fastening body 111 may be clip-coupled or slide-coupled.

The sample receiving portion 130 may have a latching protrusion 132 disposed below the inner circumferential surface. The stopping protrusion 132 protrudes from the inner wall of the sample accommodating portion 130 to form a step. At least a part of the side wall part 113 and the cut-out part 114 and the sealing sheet 115 may be positioned in the sample accommodating part 130 in a state where the cap part 110 and the sample accommodating part 130 are engaged. The lower surface of the inner wall of the sample accommodating portion 130 and the lower surface of the pressing portion 112 as well as the outer wall of the side wall portion 113 and the lower surface of the sealing sheet 115 define the second inner space S2 of the sample container 100 . The second inner space S2 can form a completely isolated space from the outside in a state where the cap portion 110 and the sample receiving portion 130 are fastened together. A specimen (not shown) may be stored in the first inner space S1 and the second inner space S2 separated from the first inner space S1. The second internal space S2 can be configured to be opened and closed by fastening and releasing between the sample accommodating portion 130 and the fastening body 111. [

In some embodiments, the sample container 100 may further include a slit portion 150 coupled with a lower portion of the cap portion 110 and positioned within the sample receiving portion 130. The slit part 150 may be located on a path along which the bacteria decay reagent CP accommodated in the first inner space S1 moves toward the second inner space S2. The engaging protrusion 132 abuts on the slit portion 150 to stably support the slit portion 150 in a state where the cap portion 110 and the sample receiving portion 130 are engaged.

Hereinafter, the slit portion 150 will be described in detail with reference to FIG. 6 is a perspective view of the slit portion of Fig.

1 to 6, the slit part 150 includes an annular slit body 151 having an inner diameter larger than the outer diameter of the side wall part 113 of the cap part 110, First slits 152a which are formed in a substantially radial manner on the first plate 152 and a second plate 153 which protrudes further downward from the inner edge of the first plate 152, Two slits 153a and a plurality of partition walls 154 disposed between the first plate 152 and the second plate 153 and protruding upward.

The slit body 151 may be a portion that is abutted against and engaged with the cap portion 110. For example, the annular slit body 151 may be reversibly coupled with the lower end of the side wall portion 113 of the cap portion 110. In the exemplary embodiment, the inner circumferential surface of the slit body 151 and the outer circumferential surface of the side wall portion 113 can be fitted together. However, the present invention is not limited thereto, and in another embodiment, the slit body 151 and the side wall portion 113 may be screwed, clipped, or slidably coupled.

The first plate 152 may be annularly arranged on the lower side of the inner circumferential surface of the slit body 151. The first slits 152a may be formed in the first plate 152 so as to be substantially radial so as to provide a path through which the bactericide reagent CP and air move.

The engaging protrusions 132 of the sample accommodating portion 130 are in contact with the slit body 151 and the first plate 152 at least partially so that the cap portion 110 and the sample accommodating portion 130 are engaged with each other. have. That is, the height of the latching jaw 132 is formed at the same level as the lower surface of the first plate 152, and the slit portion 150 can be stably supported.

The second plate 153 may protrude downward from the inner edge of the first plate 152 and may be arranged in a circular shape. The second plate 153 may have a step with the first plate 152. The upper surface of the second plate 153 may be positioned lower than the lower end of the cut-out portion 114 when the cut-out portion 114 moves to the maximum. The second slit 153a is formed in the second plate 153 so as to be substantially radial so as to provide a passage through which the bactericide reagent CP and the air move. The lower surface of the second plate 153 may be located at a level lower than the height of the engaging protrusion 132.

A plurality of partition walls 154 may be disposed between the first plate 152 and the second plate 153. The plurality of partition walls 154 are spaced apart from each other to partially separate a space above the first plate 152 and a space above the second plate 153, It is possible to make the space on the upper portion 153 of the valve body 153 in fluid communication with each other. At the same time, the partition wall portion 154 can guide the movement of the cut-out portion 114 when the cut-out portion 114 moves downward.

In the exemplary embodiment, the second slits 153a located at the center of the first slits 152a are positioned in the second internal space S2 in the first internal space S1, Lt; RTI ID = 0.0 > a < / RTI > In this case, the second slits 153a may provide a path for the air in the second internal space S2 to move into the first internal space S1. Thus, it is possible to prevent the first slits 152a and the second slits 153a having a small size from being clogged by the bacteria killing reagent CP. Also, when the amount of the bactericidal reagent (CP) allowed by the passage provided by the first slits 152a moves, a part of the bactericidal reagent CP overflows through the second slits 153a. That is, the second slits 153a may provide a secondary path in which the bactericidal reagent CP moves from the first internal space S1 to the second internal space S2.

In addition, when the specimen stored in the sample storage part 130 has a viscosity such as human sputum, the sputum does not completely mix with the bactericide (CP) . However, the sputum and bactericidal reagent CP can be more easily dispersed and mixed by the plurality of first slits 152a and the second slits 153a formed in the slit part 150, It can be expected that the terial is completely killed and the infectivity is eliminated.

In an exemplary embodiment, preparing a sample container containing a specimen having a bactericidal reagent and a bacterium not infectious to infectivity (S110) may include preparing a specimen container according to the present embodiment (S111) (S112) of receiving a sample having a bacterium that has not been detected by the cap portion in the sample holding portion disassociated from the cap portion, and combining the cap portion and the sample holding portion (S113).

7 is a view showing a step S112 of receiving a specimen K1 having a bacterium with no infectivity removed in the specimen accommodating portion 130. Figure 8 is a view showing a step S112 in which the cap portion 110 and the specimen accommodating portion 130 are combined (Step S113).

Referring to FIGS. 1 and 7, a specimen K1 having bacteria that has not been infected is provided in the specimen container 130 of the specimen container 100 (S112). For example, the specimen (K1) may be the sputum of a patient suspected of having tuberculosis. The step S112 of receiving the specimen K1 having the infectivity-free microbes in the specimen accommodating section 130 is performed by inserting the specimen K1 into the specimen accommodating section 130 in a state of being disengaged from the cap section 110 Or the like.

1, 7, and 8, the cap 110 and the sample receiver 130 are coupled and sealed (S113). In the exemplary embodiment in which the cap portion 110 and the sample accommodating portion 130 are screwed together, the second inner space S2 is defined by the screw coupling and the second inner space S2 is defined in the sample container 100, Can be kept completely isolated from the outside. In the step S113 in which the cap portion 110 and the sample accommodating portion 130 are sealed and sealed, the specimen K1 may be in a state having infectivity-free microorganisms.

(K1) having a sealed first internal space (S1) and a non-infectious microorganism accommodated therein and being separated from the first internal space (S1) and being configured to be openable and closable The sample container 100 including the second inner space S2 can be prepared (S110).

The sample container 100 according to the present embodiment is a sample container 100 in which the cap portion 110 and the sample container portion 130 are firmly engaged with each other and the contact with the outside is completely blocked 2 inner space S2 can be formed. That is, the specimen K1 housed in the second inner space S2 can be stored and / or transferred in a completely isolated state from the outside. Thus, it is possible to prevent the Mycobacterium of the specimen (K1) of the patient suspected of having the Mycobacterium tuberculosis from being propagated to the outside.

Next, the step (S120) of contacting the specimen having the bacteria-killing reagent and the bacteria having no infectivity removed will be described.

9 is a view showing a step (S120) of contacting a specimen K1 having a bactericidal reagent (CP) with a bacterium not infectious.

Referring to FIGS. 1 and 7 to 9, a step S120 of contacting a specimen K1 having a bactericidal reagent CP with a specimen K1 not infecting the infectious agent is performed in a first internal space S1 may be injected into the second inner space S2 of the sample container 100 so that the bacteria killing reagent CP and the sample K1 are contacted with each other. That is, the specimen K1 and the bacteriostatic reagent CP can be contacted in the closed second internal space S2.

In an exemplary embodiment, injecting a bactericidal reagent CP contained in the first internal space S1 of the sample container 100 into the second internal space S2 of the sample container 100 (S120) Step S121 of applying a downward pressure to the container pressing part 112 and step 122 of cutting the sealing sheet 115 partially by the cutout part 114. [

When the downward pressure is applied to the pressing part 112 of the sample container 100, the cut part 114 connected to the elastic dome and the pressing part 112 of the pressing part 112 can move downward. The lower end of the cutout portion 114 moved downward can partially cut off the approximately central portion of the sealing sheet 115. [

Which is accommodated in the first internal space S1 formed by the first internal space S1 of the cap portion 110, specifically, the side wall portion 113, the pressing portion 112 and the sealing sheet 115, The sealing sheet 115 can be partially cut off and simultaneously moved downward by gravity. For example, the bactericide reagent (CP) contained in the first internal space (S1) passes through the slit part (150) and the second internal space (130) in the sample containing part (130) S2. ≪ / RTI > The specimen K1 may come into contact with the bactericidal reagent CP in the second internal space S2 so that the bacteria of the specimen K1, such as mycobacteria, may be at least partially killed.

Next, a description will be given of a step (S130) of forming a specimen having bacteria with infectivity removed by mixing a specimen and a fungicide reagent.

10 is a view showing a step (S130) of forming a specimen (K2) having infectivity-deficient bacteria by mixing a specimen with a fungicide reagent.

Referring to FIGS. 1 and 7 to 10, the specimen is mixed with the anticancer reagent to form a specimen K2 having infectious bacteria (S130). The step (S130) of mixing the specimen with the anticancer reagent may be performed in the first internal space S1 and / or the second internal space S2 of the sample container 100. Thereby, the specimen container 100 containing the specimen K2 having the infectious organism without the cell wall being destroyed can be prepared.

In the step S120 of contacting the specimen K1 with the bacteria killing reagent CP and the specimen K1 not infecting the infectious agent and the step K1 forming the specimen K2 with the infectious bacteria removed, The cap portion 110 of the sample holder 130 and the sample holder 130 may remain coupled. In this case, the specimen (K1) having a bacterium with no infectivity removed and the specimen (K2) having a bacterium with infectivity removed may be kept blocked from the outside air.

According to this embodiment, the step of contacting the specimen (K1) having the infectivity-free microbes with the bacteria killing reagent (CP), the step (S120) of contacting the specimen K1 with the infectivity- The sample K1 and the bacteria killing reagent CP can be brought into contact with each other without opening the second internal space S2 in which the sample K1 is stored in step S130. That is, the specimen which is provided from the patient and once sealed in the second internal space S2 can be kept completely blocked from the outside air until the infectious agent is removed by the bacteria killing reagent (CP).

In an exemplary embodiment, the method further includes a step S200 of opening a space in which the specimen K2 is accommodated after the step S130 of preparing the specimen container 100 containing the specimen K2 having the infectious bacteria removed therefrom .

11 is a view showing a step S200 of releasing the coupling between the cap part 110 and the sample accommodating part S130 and opening the internal space.

Referring to FIGS. 1 and 7 to 11, the coupling between the cap 110 and the sample receiver 130 is released (S200). The inner space formed by the coupling between the cap part 110 and the sample receiving part 130 may be opened and exposed to the outside as the cap part 110 and the sample receiving part 130 are separated. That is, the specimen K2 having the infectious bacteria in the internal space can be exposed to the outside air.

On the other hand, the step (S300) of applying the pretreated sample to the glass may be a step of smearing the specimen having the infectious bacteria removed by the automatic smearing apparatus into the glass.

In an exemplary embodiment, step S300 of applying the pretreated specimen to a glass includes preparing a smear processing container including a filter membrane and an smectic membrane (S310), forming a floc (S330) of inserting the specimen having the infectious bacteria (S340), inserting the specimen having the infectious bacteria (S340), inserting the specimen having the infectious bacteria into the automatic smearing apparatus A step S350 of inserting a glass between the porous membrane and the smear membrane, and a step S360 of smearing a sample on the glass.

First, a step S310 of preparing a smear processing vessel including a filter membrane and an smelting membrane will be described in detail.

12 is a perspective view showing the smear processing container 200 used in the smear processing method according to the present embodiment. 13 is an exploded perspective view of the smear processing container 200 of FIG. 14 is a sectional view of the smear processing container 200 of Fig.

12 to 14, the smear processing container 200 according to the present embodiment includes a smear processing container body 210, a filter membrane 230 inserted into the smear processing container body 210, A smear membrane 250 inserted into the body 210 and a drain container 270 inserted into the smear processing body 210.

The smear processing vessel body 210 may have an internal space S3 in which the sample solution can be received. For example, the smear processing vessel body 210 may be a cylindrical vessel.

The discharge vessel 270 may be coupled to the lower end of the smear processing vessel body 210. For example, the discharge vessel 270 can be screwed, clipped, or slidably coupled to the smear processing vessel body 210. The discharge vessel 270 can provide a space in which the residual solution of the sample that has passed through the filter membrane 230 is received. A plurality of discharge holes 270a may be formed on the upper surface of the discharge vessel 270 to allow the flow of the fluid.

The filter membrane 230 may be inserted and disposed within the body of smear processing vessel 210. The filter membrane 230 may comprise a porous membrane. The filter membrane 230 can separate foreign matter or mucus larger than the bacteria or cells to be smoked in the specimen K2 having the above-mentioned infectious bacteria. This makes it possible to more smoothly form the surface of the specimen smeared on the slide.

The smear membrane 250 may be inserted into the smear processing vessel body 210. The smear membrane 250 may be disposed between the filter membrane 230 and the drain vessel 270. The smear membrane 250 may be disposed directly on the discharge vessel 270 and supported by the upper surface of the discharge vessel 270.

The sample filtered by the filter membrane 230 can be adsorbed to the smear membrane 250. The smear membrane 250 may be made of a porous membrane. The size of the pores of the smear membrane 250 can be appropriately selected depending on the size of the bacteria to be detected. That is, at least a portion of the bacteria in the filtered specimen may be adsorbed to the smear membrane 250. Residual solutions containing unnecessary cells and moisture in the filtered specimen can also be passed through the smear membrane 250 and into the drain container 270 side.

Fig. 15 is a view showing a step (S320) of injecting a specimen K2 having infectious bacteria into the internal space of the smear processing container 200. Fig. 16 is a view showing a step (S330) of inserting the smear processing container 200 into the automatic smashing apparatus 300. [ 17 is a diagram showing a step (S340) of filtering a specimen. 18 is a view showing the step of inserting the glass 400 between the filter membrane 230 and the smear membrane 250 (S350). 19 is a view showing a step (S360) of smearing a specimen on a glass (400). Fig. 20 is a view showing a glass 400 in a state in which bacteria are smoothed.

First, referring to FIGS. 1 and 15, a specimen K2 having infectious bacteria removed is injected into the internal space S3 of the smear processing container 200 (S320).

1, 15 and 16, the smear processing container 200 containing the specimen K2 is inserted into the insertion port 330 of the automatic smearing apparatus 300 (S330). The automatic smearing apparatus 300 may include an operation unit 310 and an insertion port 330. The operation unit 310 may be a part for controlling the operation of the automatic smudging apparatus 300. The insertion port 330 may have a size and a shape corresponding to the smear processing container 200 according to the present embodiment.

1 and 15 to 17, the sample is filtered (S340). Step S340 of filtering the specimen K2 drains the specimen K2 having the infectious bacteria contained in the smear processing container 200 to the drain container 270 side, May be the step of inducing permeation of the membrane 230. Unnecessary solid foreign substances or mucus components in the specimen K2 having infectious bacteria removed in step S340 are filtered by the filter membrane 230 and only the cells and / or bacteria pass through the filter membrane 230 And may be adsorbed to the smear membrane 250. The step of filtering the specimen (S340) may be performed by the automatic smearing apparatus 300 inside the automatic smear apparatus 300.

1 and 15 to 18, a glass 400 is inserted between the filter membrane 230 and the smear membrane 250 of the smear processing vessel 200 (S350).

Step S350 of inserting the glass 400 includes a step S351 of separating the smear processing vessel body 210 from the smelting membrane 250 and the discharge vessel 270 and a step of removing the smear membrane 250 and the filter membrane 230, And interposing a glass 400 therebetween (S352). In this case, solid foreign substances and / or mucus substances are filtered on the filter membrane 230, and the smear membrane 250 is in a state in which the cells of interest, background cells and / or microorganisms (such as mycobacteria) Residual solutions containing unwanted micro cells and / or moisture may be contained within the discharge vessel 270. The step of inserting the glass 400 (S350) may be performed by the automatic smearing apparatus 300 inside the automatic smear apparatus 300. [

Next, referring to FIGS. 1 and 15 to 19, a specimen is laid on a glass 400 (S360). The glass 400 sandwiched between the smear membrane 250 and the filter membrane 230 has a first surface (upper surface in the drawing) facing the filter membrane 230 and a second surface Upper surface). The step S360 of applying the specimen to the glass 400 may be performed by contacting the second surface of the glass 400 with the smear membrane 250 so that the cells and / To the second surface (the lower surface in the figure) of the glass 400. In this step,

Referring to FIG. 20, the smear sample K3 may be formed on the second surface (upper surface in the drawing) of the glass 400.

On the other hand, although not shown in the figure, specimens having infectious bacteria removed can be stained on a slide, and dyeing and microscopic examination can be performed. The staining can be carried out through anti-acid staining, such as the Ziehl-Neelsen method or fluorescence staining. In particular, according to the antifungal agent and the smear treatment method using the microbacterium according to the present embodiment, only the microbacterium is selectively killed, while the microbacterium-specific microbacterium is maintained in a visually recognizable state, And the cell wall of other bacteria in the sample may not be destroyed. This makes it possible to perform contrast dyeing of background cells except for mycobacterium and to enable clear visual identification through a microscope.

In addition, the specimen can remain unexposed to the outside air until the infectivity of the Mycobacterium in the specimen collected from the patient suspected of having the Mycobacterium tuberculosis is removed. This makes it possible to prevent the infection of the tuberculosis of the inspector who performs the tuberculosis test in advance.

Hereinafter, an experimental example of the present invention will be described.

[ Manufacturing example ]

A composition was prepared comprising 4.0% by weight of sodium hydroxide (NaOH), 2.9% by weight of sodium citrate, 3.0% by weight of sodium hypochlorite (NaOCl) and the remaining amount of distilled water.

[ Comparative Example  One]

The composition was prepared in the same manner as in the above preparation example except that the sodium hypochlorite content was adjusted to 6.0 wt% and the remaining amount of distilled water was included.

[ Comparative Example  2]

A composition was prepared in the same manner as in the above Preparation Example except that the sodium hypochlorite content was adjusted to 9.0 wt% and the remaining amount of distilled water was included.

[ Experimental Example ]

The viscosity and filtering characteristics of the compositions according to the Preparation Examples and Comparative Examples 1 and 2 were evaluated and the results are shown in Table 1 below. In Table 1, & cir & indicates that the viscosity is low or the filtering characteristic is excellent, & cir & indicates the viscosity is high or the filtering characteristic is bad, and &

The compositions according to the above Preparation Examples and Comparative Examples 1 and 2 were mixed with sputum of tuberculosis patients, respectively, and the mixture was applied to a slide and stained. The results are shown in Table 1 below. In the following Table 1, & cir & indicates that the dyeing condition is excellent and the detection of mycobacteria is easy, X means that the dyeing condition is poor and the detection of mycobacteria is not easy, and &

Evaluation of composition characteristics Evaluation of dyeing characteristics Viscosity Filtering characteristics Background cell status after smear Image Rating Manufacturing example ○ △

○ Comparative Example 1 △ ×

△ Comparative Example 2 × ○

×

While the present invention has been described with reference to exemplary embodiments, it is to be understood that the invention is not limited to the disclosed exemplary embodiments, but many variations and modifications may be made without departing from the spirit and scope of the invention. It will be appreciated that many variations and applications not illustrated above are possible. For example, each component specifically shown in the embodiments of the present invention can be modified and implemented. It is to be understood that all changes and modifications that come within the meaning and range of equivalency of the claims are therefore intended to be embraced therein.

100: sample container
200: smear processing container
300: Automatic smearing device
400: Glass

Claims (8)

(a) preparing a sample container including a cap portion containing a bacteriostatic reagent containing sodium hydroxide, sodium hypochlorite, sodium citrate and a solvent, and a sample accommodating portion configured to be able to engage with the cap portion;
(b) receiving the specimen in a space inside the specimen accommodating portion in a state in which the cap portion is separated from the specimen accommodating portion;
(c) sealing the specimen by coupling the cap portion with the specimen accommodating portion accommodating the specimen;
(d) injecting the germicidal reagent contained in the cap part into a space containing the specimen in a state in which the specimen is sealed, thereby bringing the germicidal reagent into contact with the specimen; And
(e) separating the cap portion and the sample receiving portion again, and laminating the sample to a glass,
Until the step (e) is performed after the step (c), the specimen is kept isolated from the outside of the specimen container,
Wherein the sample container further includes a slit portion disposed between the cap portion and the sample containing portion,
The slit portion
Annular slit body,
A first plate protruding downward from an inner circumferential surface of the slit body and having a plurality of slits,
A second plate further protruding downward from an inner edge of the first plate and having a plurality of slits with a step,
And a plurality of protruding partition wall portions disposed between the first plate and the second plate and projecting upward.
A method for smear treatment of a specimen. delete delete The method according to claim 1,
The cap unit includes:
An annular fastening body having an engaging means capable of engaging with the sample accommodating portion,
A pressing portion disposed on an upper side of the inner circumferential surface of the annular fastening body,
A side wall portion protruding downward from the edge of the pressing portion,
A cutout portion protruding downward from the center of the pressing portion and inserted and disposed in the side wall portion,
And a sealing sheet disposed at a lower end of the sidewall portion and forming a space for accommodating the fungicide reagent together with the pressing portion and the sidewall portion,
The step of contacting the specimen with the anti-
Applying a downward pressure to the pressing portion and partially cutting the sealing sheet so that the cut portion protruding downward from the pressing portion injects the bacteria killing reagent accommodated in the cap portion into the space containing the specimen, . delete The method according to claim 1,
The step of contacting the specimen with the bactericide reagent is a step of removing the infectivity of the specimen contained in the specimen,
Wherein the specimen remains in a state of being shielded from the outside air during the process of eliminating the infectivity of the germ of the specimen. The method according to claim 6,
Wherein the step of applying the specimen to the glass is a step of smearing the specimen having the infectious bacteria removed by the automatic smashing apparatus into a glass. 8. The method of claim 7,
The step of applying the specimen having the infectious bacteria removed to the glass using the automatic smearing apparatus comprises:
Preparing a smear processing vessel including an smear processing vessel body having an inner space, a filter membrane inserted in the smoothing processing vessel body, and an smelting membrane inserted in the smoothing processing vessel body and disposed below the filter membrane, ,
A specimen having infectious bacteria contained in the specimen accommodating section and injecting the remaining antifungal reagent into the internal space of the smear processing container,
Inserting the smear processing container containing the specimen having the infectious bacteria removed therefrom and the remaining antifungal reagent into the automatic smearing apparatus,
Filtering the specimen with the infectious bacteria removed using the filter membrane,
Inserting the glass between the filter membrane and the smear membrane, the method comprising: inserting a glass having a first surface opposite the filter membrane and a second surface opposite the smectite membrane;
And smearing the specimen on the second side of the glass by bringing the smear membrane into contact with the second side of the glass.

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