JP2001095559A - Assisting agent for centrifugal collection of microorganism, recovery of microorganism and recovery of acid-fast microorganism - Google Patents

Abstract

PROBLEM TO BE SOLVED: To develop the application field of a coagulant such as an aluminum polychloride used as a cleaning agent for the removal of clouding substance from water in the centrifugal separation and recovery of microorganisms from a stock solution or a specimen based on the finding that the recovery rate of the microorganism can be improved at the same centrifugal force by the use of the coagulant and to provide a method for recovering microorganisms in high yield by the addition of the agent or chemical in conventional centrifugal separation process. SOLUTION: Objective microorganisms are recovered from a precipitate by diluting a stock solution of specimen to pH 7 or thereabout, adding 0.01-0.05 mL of an aqueous solution containing the optimum amount of a coagulant such as aluminum polychloride to 10 mL of the diluted solution, stirring the mixture, leaving at rest for certain period and separating by centrifugal separation at a centrifugal force of 1,300×g to 1,800×g.

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001]

TECHNICAL FIELD The present invention relates to an auxiliary agent for efficiently recovering a target bacterium from an undiluted solution or a specimen or a method for recovering a bacterium using this auxiliary agent. The present invention relates to a method for efficiently collecting acid bacteria. In addition, the auxiliary agent according to the present invention includes a use invention which has been conventionally developed as a bacterial coagulation auxiliary agent, which has been conventionally used as a purifying agent for drinking water, industrial waste liquid, and the like.

[0002]

2. Description of the Related Art Conventionally, as a method for recovering this kind of bacteria, a method of recovering the bacteria by centrifugal sedimentation is mainly used. However, as a method for improving the efficiency of collecting bacteria, it is only considered to increase the centrifugal force. . Increasing the centrifugal force not only increases the size of the device but also increases the cost of the equipment and reduces its usefulness. If the centrifugal force is excessively increased, the target bacteria may be killed, and increasing the centrifugal force naturally has a limit. Although electrophoresis has been used in some cases since about 1960, the technique is complicated and the apparatus itself is expensive, and this method is not a method for improving the bacterial yield.
A type of polyaluminum chloride, aluminum sulfate, ammonium alum, potassium alum, ferrous sulfate, ferric sulfate, anionic, nonionic and cationic types used when purifying drinking water and industrial waste liquids At least one of the high molecular organic flocculants consisting of
It is mixed in the treated water, turbid substances and the like are settled and sieved, and the supernatant is supplied to drinking water or miscellaneous water. The turbid substances and the like that have been sieved are discarded after incineration or sterilization. Therefore, these polyaluminum chlorides and the like are used as agents for purifying water and the like.
I could not find it at all.

[0003]

According to the present invention, when the target bacterium is recovered by centrifugation from a stock solution or a specimen, if polyaluminum chloride or the like which has been conventionally used as a water purifying agent is used, the centrifugal force is the same as that of the prior art. However, it was found that the recovery rate of bacteria was significantly improved, and the use of the drug or chemical was developed, and the method of adding this drug or chemical to a conventional centrifugation method was used to increase the rate of bacteria. To provide the market with an efficient collection method. In particular, the present invention provides a method for detecting a trace amount of mycobacteria from sputum.

[0004]

In order to achieve the above-mentioned object, the invention of the present invention relates to a first coagulant mainly composed of polyaluminum chloride, aluminum sulfate, ammonium alum, potassium alum, ferrous sulfate, sulfuric acid. A centrifugal collector characterized in that at least one coagulant of at least one of a second coagulant composed of at least one of ferric iron, a high molecular organic coagulant composed of at least one of anionic, nonionic and cationic is used as a main component. Bacterial aid.

In order to achieve the above object, a related method invention includes a 25-50% aqueous solution of a first coagulant containing polyaluminum chloride as a main component, aluminum sulfate, ammonium alum, 0.5 to 10% aqueous solution of a second flocculant comprising at least one of potassium alum, ferrous sulfate and ferric sulfate; and 0 to 0 of a high molecular organic flocculant comprising at least one of anionic, nonionic and cationic. 0.05 to 0.3%
At least one kind of the aqueous solution is added at a ratio of 0.01 to 0.05 ml, and the mixture is stirred, and after a while, centrifuged to collect the target bacteria from the sediment.

In order to achieve the above object, as a related method invention, a first step of adjusting the stock solution of the specimen to a pH around the optimum pH of the target bacterium with a buffer solution. 25% to 50% aqueous solution of a first coagulant containing polyaluminum chloride as a main component per at least 10 ml of the treatment liquid of the first step, and at least one of aluminum sulfate, ammonium alum, potassium alum, ferrous sulfate and ferric sulfate. 0.5 of the second flocculant
To 10% aqueous solution, and 0.05 to 0.3% aqueous solution of at least one coagulant of at least one high molecular organic coagulant comprising at least one of anionic, nonionic and cationic. 01 to 0.05
The mixture is added at a rate of milliliter, stirred, and after a while, centrifuged and centrifuged to recover the target bacteria from the sediment.

In order to achieve the above object, the centrifugal force of another method for recovering bacteria has a centrifugal force of 1300 × g to 1800 × 1800.
Xg (2600 rpm in a 16 cm radius centrifuge)
(Equivalent to about 3150 rpm) for about 25 to 15 minutes. In order to achieve the above object, as another method invention, the sample stock solution of the method for collecting bacteria may be characterized in that it contains at least one kind of useful edible bacteria and medicinally useful bacteria. .

[0008] In order to achieve the above object, another method invention may be characterized in that the sample stock solution in the method for collecting bacteria is a sample stock solution containing pathogenic bacteria. In order to achieve the above object, as another method invention, a sputum presumed to contain acid-fast bacterium was first treated with a disinfectant such as an alkali, and then sterilized with a homogenous bacterium other than the acid-fast bacterium. The first step to be performed. The second step of adjusting the solution adjusted in the first step with a buffer solution having a pH of about 7. The second process is composed of 25 to 50% aqueous solution of the first flocculant containing polyaluminum chloride as a main component, aluminum sulfate, ammonium alum, potassium alum, ferrous sulfate, and ferric sulfate per 10 ml of the treatment liquid in the second step. 0.5 to 10% aqueous solution of a second flocculant, 0.05 to 0.3 of a high molecular organic flocculant comprising at least one of anionic, nonionic and cationic ones
% Aqueous solution at a rate of 0.01 to 0.05 ml, and after stirring for a while, centrifugal force of 1300 × g to 1800 × g for about 25 to 15 minutes A method for recovering an acid-fast bacterium, comprising centrifuging and recovering a target acid-fast bacterium from a sediment.

The radius of the aforementioned centrifuge is not limited to 16 cm, and as a result, the acceleration of gravity is about 1
Any method can be used as long as it is performed using a device capable of generating one centrifugal force in the range of 300 × g to 1800 × g. preferable. If the radius is smaller than 16 cm, the rotation speed will be faster than 3000 rpm, and if the radius is larger than 16 cm, the rotation speed will be 3000 rpm.
rpm, and such cases are included in the embodiments of the invention (see the graph of FIG. 1).

Action of the Invention The specific invention described in claim 1 is an invention of a product. Generally, the surface of a cell is charged with negative ions and repels each other, and fine bacteria are stable in a floating state. Therefore, even if it is used for centrifugal force collection as it is, sufficient centrifugal force does not act on fine cells, and it is not possible to efficiently collect bacteria.
Therefore, containing the target bacteria, prior to collecting the centrifugal force from the sample liquid whether or not other bacteria are contained, the three aqueous solutions containing the flocculant at a predetermined ratio in the target sample stock solution When at least one of the aqueous solutions is added,
This neutralizes the surface potential of the bacterium due to the positive charge. Thus, the cells are in a state where they can easily approach each other, and the bacteria and other contaminants are mutually aggregated to form an aggregate. In this state, if a centrifugal force is applied, it effectively acts on the aggregates, precipitates and separates, and the probability of cells being collected in the precipitates is significantly improved.

In the invention of claim 2, first, when the sample stock solution is not viscous, at least one aqueous solution among the three aqueous solutions of the aggregating agent is added to the undiluted solution. Mix thoroughly to neutralize each cell surface potential evenly and form larger aggregates. If the mixing ratio of each aqueous solution is less than the above range, a sufficient neutralizing effect on the surface potential of each bacterial cell cannot be obtained, and if it is too large, it will be positively charged and aggregation will not occur again. In such a state, when a centrifugal force is applied to the same degree as in the related art, the aggregated bacteria and other contaminants are easily precipitated by centrifugation.

According to the third aspect of the present invention, when the sample stock solution is highly viscous or when some processing is performed, such as when it is collected from a culture medium, the buffer solution,
Alternatively, if the sample stock solution is adjusted to a pH around the target bacterium with a diluent, the viscosity of the sample stock solution is reduced, stirring and fluidity are improved, the survival rate of the target bacterium is increased, and the sample is added in the second step. At least one of the three aqueous solutions is sufficiently mixed to neutralize the surface potential of each bacterial cell evenly, and has the effect of forming a larger aggregate.

According to the fourth aspect of the present invention, since the centrifugal force and the operation time are within the above-mentioned ranges, an apparatus generally used as a centrifuge for separating bacteria or the like can be used as it is. The rotation speed is not high enough to destroy the target cells during the separation, and the operation time is short and the operation is efficient. Therefore, the sediment serves to recover a high percentage of bacteria.

According to the fifth aspect of the present invention, in addition to the effects of the second, third or fourth aspect of the present invention, useful food bacteria,
It has the function of collecting bacteria useful for medicine at a high rate. In the invention according to claim 6, the second, third or fourth aspect of the present invention is provided.
In addition to the effects of the described invention, when detecting an unspecified pathogenic bacterium, it has an effect of increasing the rate at which the bacterium can be detected from a small sample because of its high collection efficiency or even from a sample with a very small number of bacteria.

In the invention according to claim 7, claim 2 is provided.
Or the sputum presumed to contain the acid-fast bacterium in addition to the action of the invention according to 3 as a sample stock solution, first with a disinfectant such as alkali,
Because the bacteria other than the acid-fast bacterium are sterilized and homogeneous,
Only the acid-fast bacterium is to be collected and further diluted with a buffer solution having a pH of about 7, so that the consistency of the sample stock solution is reduced and aggregation is likely to occur. Also, when it contains an acid-fast bacterium, it has an effect of hardly lowering the survival rate. Other actions are the same as the actions of the inventions of claims 2 and 3, and the acid-fast bacterium is efficiently recovered in the end, or the acid-fast bacterium can be discovered even from a sample stock solution containing a trace amount of the acid-fast bacterium. Performs a possible function.

Embodiment 1 This is an embodiment of the invention as set forth in claims 2, 3, 4 and 5 using the auxiliary of the invention as set forth in claim 1, wherein the cells contain a kind of useful edible bacteria. The subsequent liquid medium was adjusted to the optimum pH of the target bacterium, for example, pH 7, with a buffer solution.
A 25 to 50% aqueous solution of polyaluminum chloride as a first coagulant is added to the milliliter in an amount of 0.01 to 0.05 milliliter, and after stirring for a while, 1300 × g
Centrifuge at 1800 × g for 25 to 15 minutes.
Instead of the 25 to 50% aqueous solution of the first flocculant, a 0.5 to 10% aqueous solution of the second flocculant or 0.05 to 0.3% of at least one flocculant among the high molecular organic flocculants An aqueous solution may be used. Thus, the target bacteria are recovered from the sediment. The buffer is not limited to a phosphate buffer, and depending on the type of bacteria to be recovered,
Use a buffer in which the pH of the buffer corresponds to the optimal pH of the target bacterium.

Embodiment 2 This is an embodiment of the method invention according to claim 2 using the assistant of the invention described in claim 1. When a relatively large amount of the sample stock solution is obtained and the liquid is less viscous, for example, in the case of patient urine,
As it is, as a sample stock solution, an aqueous solution containing the aggregating agent was added to the sample stock solution (10 ml) in the same manner as in Embodiment 1 at the ratio described above, and stirred, and after a while, 15 ml as a centrifuge tube And a centrifugal force of 1300 × g to 1800 × g.
Centrifuge for 5 to 15 minutes. Thus, from the sediment,
Collect the desired bacteria. Alternatively, the sediment is used as a culture source and transplanted into a medium.

Embodiment 3 Claims 3, 4 and 5 using the auxiliary of the invention described in claim 1.
It is an embodiment of the invention described, wherein the liquid medium after culturing containing one kind of medicinal or edible useful bacteria is diluted with a buffer solution, adjusted to the optimal pH of the target bacteria, and treated. An aqueous solution containing the aggregating agent is added to 10 ml of the liquid in the same ratio as in the first embodiment, and the mixture is stirred. After a while, the cells are centrifuged under the same conditions as in the first embodiment.

Embodiment 4 This is an embodiment including the invention according to claim 6, wherein a stock solution containing unknown pathogenic bacteria, for example, a solution of patient excrement or food residue is collected with a buffer solution to grow bacteria to be collected. The sample stock solution is adjusted to the respective pH using a buffer having an optimum pH, for example, pH 7, pH 5.6, or pH 8.0 or higher, and corresponding pHs are different. The same aqueous solution containing the aggregating agent as in Example 1 is mixed with each of the above-described ratios, and the cells are collected by centrifugation under the same conditions as in Embodiment 1. After centrifugation, this is further cultured using this as a culture source to identify the bacteria.

Embodiment 5 An embodiment including the invention according to claim 7, wherein sputum which is presumed to contain acid-fast bacilli, especially tuberculosis bacterium, is used as a sample stock solution, and the anti-acid bacterium is firstly treated with a disinfectant such as an alkali. A first step in which bacteria other than bacteria are sterilized and made uniform. The solution adjusted in the first step is diluted and adjusted with a buffer solution having a pH of about 7. by this,
The second to correct alkalinity by disinfectants such as alkali
Process. The same aqueous solution containing the flocculant as in the first embodiment was mixed with the above-mentioned ratio in 10 ml of the treatment liquid in the second step at the above-mentioned ratio, and each was centrifuged under the same conditions as in the first embodiment to obtain a sediment. The target acid-fast bacterium is recovered from the inside. Alternatively, the sediment is used as a culture source, and further cultured in a liquid medium for a predetermined period of time, and the same method is repeated again to collect mycobacteria, particularly tuberculosis bacteria.

As the buffer used in the present invention, a phosphate buffer (including Slensen's phosphate buffer) is used, and the pH is previously determined and prepared according to the purpose. Physiological saline or purified water is used as the diluting solution.

[0022] peptone 4% as an experimental example Experimental bacteria Tokyo BCG strains sputum model, using a 1% aqueous solution mucin, which put the bacteria 10 ^-4 mg / ml, which was used as a specimen. One milliliter of the sample and 9 milliliters of M / 15 phosphate buffer were added to each of three centrifuge tubes. One of the centrifuge tubes was left as it was, and the other was mixed with the 40% aqueous solution of polyaluminum chloride as an adjuvant. After stirring, centrifugal separation was performed using a centrifugal separator with a radius of 16 cm at a rotation of 3000 rpm (approximately 1600 × g as indicated by the acceleration of gravity) for 20 minutes, and centrifugal separation was performed. Five milliliters of the solution were inoculated and cultured in 1% Ogawa's medium for 5 cells each, and the number of bacteria remaining in the supernatant was counted. The number of bacteria in the sediment was measured. The results are as shown in Table 1.

[0023]

Table 1 Number of supernatant bacteria Number of sediment bacteria Total number of bacteria Bacterial recovery 3000 rpm 864 1,372 2,236 61.4% 300 rpm + auxiliary 126 2,288 2,414 94.8% 3,000 xg 567 1,796 2,372 75.7

As can be seen from Table 1, only 300
By centrifugation at 0 rpm, only 61.4% of the solution was recovered. However, by adding the auxiliary agent according to the first aspect of the present invention within the above-mentioned range, the same centrifugal force and operation time were obtained.
A tremendous bacterial recovery rate of 8% was obtained, and the
75.7 centrifugal force of × g (equivalent to 4,100 rpm)
%, Which is much higher than the above.

[0025]

According to the invention of the first aspect, each of the above-mentioned flocculants is used as described above before centrifugation.
When added to the sample preparation solution, the charge of the cells is neutralized, mutual repulsion is eliminated, the cells are easily aggregated, the cells become large colonies, and a centrifugal force is exerted. Therefore, the effect of improving the bacterial recovery is exhibited.

According to the second aspect of the present invention, the method is suitable for the case where the sample stock solution is relatively large in volume and the consistency is high, and has an effect that even a very small amount of pathogenic bacteria can be found. In the method according to the third aspect of the present invention, the pH and the buffer solution and the diluting solution are adjusted in advance.
This method is a method of adding and stirring the aqueous solution to which the coagulant is added at a predetermined ratio before centrifugation, and therefore, the mixing is easy, and the flocculation of the cells is also easy. As described above, the target bacterium can be recovered at a high recovery rate, the centrifugal rotation speed can be relatively low, and there is little possibility that the cells are destroyed. In the case of the centrifugal force described in claim 4, a widely used centrifugal separator can be used, there is no possibility of destroying the cells, the operation time is not too long, the workability is good, and the efficiency of collecting bacteria is particularly high. Good outside.

In the method according to the fifth aspect of the present invention, since the above-mentioned method is used for separating useful food bacteria and medicinal useful bacteria as specimens, the recovery rate of these bacteria is improved, and This has the effect of improving the production efficiency of bacteria and medicinal useful bacteria. In the method invention according to claim 6, as the specimen,
Using excretions or other body fluids of patients who may contain some pathogenic bacteria, preserved foods, food residues, etc., centrifugation after using the above-mentioned auxiliary, and using this as a culture source, a higher concentration culture It has the effect of facilitating the discovery of pathogenic bacteria.

In the method according to the present invention, the sputum of a patient presumed to contain mainly acid-fast bacilli as a specimen is first disinfected with an alkaline disinfectant, thereby obtaining a sample other than acid-fast bacilli. Since the bacteria are killed and then returned to near neutrality again with the phosphate buffer, the acid-fast bacteria are rarely killed or diminished during the subsequent method. Therefore, if acid-fast bacterium is contained, it is recovered at a high recovery rate, so that what was conventionally false positive can be clearly detected, and subsequent treatment or isolation can be performed quickly, and the patient himself It is a gospel that hastened recovery and reduced the prevalence of the disease socially.

[0029] In the invention according to claims 2 to 7,
If the amount of the polyaluminum chloride as the auxiliary agent is also in the above-mentioned range, a sufficient effect of aggregating cells can be obtained. If the amount exceeds this range, the effect of aggregating cells may be hardly obtained.

Embodiment 1 Examples of useful foods by fermentation Example 1 In the case of lactic acid bacteria Lactic acid bacteria cultured in a normal and clean medium without germs are generally considerably acidic due to lactic acid produced by the lactic acid bacteria itself, and first adjusted to pH 7 with a phosphate buffer. . This treatment solution is added to the above-mentioned aqueous solution of the first flocculant containing polyaluminum chloride at the above-mentioned ratio, and after stirring, the mixture is centrifuged at 1600 × g for 20 minutes, and lactic acid bacteria are recovered from the sediment. . As useful food bacteria, other than the above,
It can be used for recovery of various yeasts and yeasts.

2. Examples of useful chemicals by culturing Example 2 In the case of Streptomyces limosas In a clean liquid medium, a culture of Streptomyces limosas was used as a stock solution, adjusted to pH 7 with a phosphate buffer, and then similar to Example 1 The target bacteria were recovered from the sediment. In addition to the above-mentioned bacteria, other antibiotic-producing bacteria such as Penesillium can be similarly collected.

3. Example 3 Detection of Pathogenic Bacteria Example 3 In the case of pneumococcus A patient's sputum was used as a buffer having a pH contained in the optimal pH of pneumococcus. After adjusting to 1 to 8.3, the same method as in Example 1 is performed, and pneumococcus is recovered from the sediment. Alternatively, the sediment is transplanted to a clean medium, and the same method is repeated again to collect pneumococci. Example 4 In the case of B. pertussis A patient's sputum was subjected to
Adjust to 4.3 to 5.6 with phosphate buffer as in Example 3. If necessary, the present invention also includes a method of adjusting the pH to the above range with a pH adjuster. After that, the same method as in the first embodiment is performed. In addition to Examples 3 and 4, it is most suitable for detecting Gram-positive cocci such as staphylococci, Gram-positive bacteria such as diphtheria, and Gram-negative bacteria such as pathogenic intestinal bacteria other than pertussis.

Example 5 In the case where pathogenic bacteria such as food poisoning have not yet been identified, a residue of food, a part of preserved food, a part of excretion of a patient, and a kind of body fluid collected from the patient are used as a sample stock solution,
PH is adjusted to around 7 using the above buffer solution. Alternatively, buffer solutions of pH 6, pH 7, and pH 8.3 are prepared in advance, and these are used to adjust the pH of the sample stock solution to pH 6 centered on pH 7.
To 8, pH 4.3 to 6, and pH 8 to 8.5. Thus, polyaluminum chloride is added to each of the treatment solutions in the same manner as in Example 1, and the precipitates are centrifuged and the respective sediments are separately cultured. Furthermore, each cultivation was separately collected for each colony formed in the medium, and these were adjusted with a buffer solution in the same manner as the previous time, polyaluminum chloride was added, centrifuged, and the bacteria in the sediment were examined with a microscope. Inspect to detect pathogenic bacteria and identify food poisoning bacteria. Example 6 When the sample stock solution is urine In the case of urine, since the sample source solution is relatively large and the viscosity is small, immediately add an aqueous solution of polyaluminum chloride without diluting or adding a buffer solution, and centrifuge. To detect syphilis, gonococci and pathogens of urinary diseases in the sediment

Example 7 Mycobacteria, especially Mycobacterium tuberculosis The sputum of a patient determined to be infected with tuberculosis by another test is disinfected with a 2% caustic soda disinfectant to produce other bacteria or pathogenic bacteria. Sterilize. The sample, which has become alkaline once, is then adjusted to around pH 7 with a phosphate buffer. Mycobacterium tuberculosis has an optimal pH
Is from 5.0 to 8.4, so that M. tuberculosis can be kept alive. Thereafter, in the same manner as in Example 1, an aqueous solution of polyaluminum chloride is added, and the mixture is centrifuged and precipitated. In some cases, the sediment is cultured, the above-mentioned polyaluminum chloride is added to the culture, and the mixture is centrifuged and sedimented again to examine whether M. tuberculosis is present in the sediment. improves.

In each of the examples, instead of a 40% aqueous solution of polyaluminum chloride, 2% of polyaluminum chloride was used.
The same effect can be obtained by using a 5 to 50% aqueous solution, and 0.5 to 50% of a second coagulant comprising at least one of aluminum sulfate, ammonium alum, potassium alum, ferrous sulfate and ferric sulfate. 10% aqueous solution, at least one aqueous solution of 0.05 to 0.3% aqueous solution of at least one coagulant of high molecular organic coagulant of one kind of anionic, nonionic and cationic, Generally similar results are obtained. Further, a mixed aqueous solution of an aqueous solution of the first flocculant and at least one coagulant of the high molecular organic coagulant, and a mixed aqueous solution of the aqueous solution of the second coagulant and at least one coagulant of the high molecular organic coagulant are used. When used, substantially the same results are obtained.

[Brief description of the drawings]

FIG. 1 is a centrifugal force calculation monograph.

 ──────────────────────────────────────────────────続 き Continued on the front page F term (reference) 4B063 QA01 QA19 QQ03 QQ05 QQ06 QQ07 QQ16 QQ20 QR41 QR50 QR51 QS11 QS13 QS20 4B065 AA01X AA36X AA38X AA50X AA57X AA67X AA72X BD15 BD22 BD25 BA04 BA05 BA22 CA20 DA04 DA05 DA06 DA15 DA16 DB01 DC02 DC06 DC07 DC08 EA04 EA13 EA14 EA15 EA16 EA22 EA39

Claims (7)

[Claims] 1. A first coagulant comprising polyaluminum chloride as a main component, a second coagulant comprising at least one of aluminum sulfate, ammonium alum, potassium alum, ferrous sulfate and ferric sulfate, anionic and nonionic A centrifugal bacteria collection assistant characterized in that at least one kind of cationic organic polymer flocculant is used as a main agent. 2. 25 to 50% of a first flocculant containing polyaluminum chloride as a main component per 10 ml of a sample stock solution
Aqueous solution, aluminum sulfate, ammonium alum,
0.5 to 10% aqueous solution of a second flocculant comprising at least one of potassium alum, ferrous sulfate and ferric sulfate; and 0 to 0 of a high molecular organic flocculant comprising at least one of anionic, nonionic and cationic. Of at least one aqueous solution of 0.01 to 0.05%
A method of collecting bacteria at a rate of milliliters, stirring the mixture for a while, and then centrifuging to collect the desired bacteria from the sediment. 3. A first step in which a sample stock solution is adjusted to at least an optimum pH of a target bacterium with at least one of a buffer solution and a diluting solution. First
25 to 50% aqueous solution of the first coagulant of polyaluminum chloride per 10 ml of the processing solution, and the second coagulant comprising one of aluminum sulfate, ammonium alum, potassium alum, ferrous sulfate and ferric sulfate 0.
A 5 to 10% aqueous solution, a high-molecular organic coagulant comprising at least one of anionic, nonionic and cationic types.
At least one aqueous solution of the three aqueous solutions of 0.05 to 0.3% aqueous solution was added at a ratio of 0.01 to 0.05 ml, and the mixture was stirred and centrifuged for a while. Bacterial recovery method to be recovered. 4. The centrifugal force of the centrifugation is 1300 × g to 1800 × g (260 centimeters in a 16 cm radius centrifuge).
The method according to claim 2 or 3, wherein the method is carried out for about 25 to 15 minutes (equivalent to 0 to 3150 rpm). 5. The method according to claim 2, wherein the sample stock solution contains at least one kind of useful edible bacteria and useful medicinal bacteria. 6. The method according to claim 2, wherein the specimen is a specimen stock solution containing pathogenic bacteria. 7. A first step in which sputum presumed to contain acid-fast bacteria is used as a sample solution, and bacteria other than the acid-fast bacteria are sterilized with a disinfectant such as an alkali to make the solution homogeneous. A second step of diluting and adjusting the solution adjusted in the first step with a buffer solution having a pH of about 7. 25 to 50% of the first flocculant containing polyaluminum chloride as the main component per 10 ml of the second process liquid
Aqueous solution, aluminum sulfate, ammonium alum,
0.5-10% aqueous solution of a second aggregating agent comprising a kind of alum, ferrous sulfate, ferric sulfate, anionic,
Nonionic, at least one aqueous solution among three aqueous solutions of 0.05 to 0.3% aqueous solution of a high molecular organic coagulant comprising at least one of cationic type is added at a ratio of 0.01 to 0.05 ml; After stirring for a while, 1
About 25 minutes to 1 at a centrifugal force of 300 xg to 1800 xg
A method for recovering an acid-fast bacterium, comprising centrifuging for 5 minutes and recovering a target acid-fast bacterium from a sediment.