Digestive juice formula, preparation method and application of clinical microbiological test room mucous specimen

Abstract

The invention belongs to the technical field of medical reagents, and particularly relates to a digestive juice formula of a clinical microbiological laboratory mucous specimen, a preparation method and application thereof. The formula of the digestive juice comprises the following components in concentration ratio: 0.0125%-0.2% (w / v) of type I collagenase; 0.0125%-0.2% (w / v) of EDTA (Ethylene Diamine 0.01526%-0.25% (w / v) of trypsin; 0.02%-0.32% (w / v) of DNA enzyme; 0.02%-0.32% (w / v) of hyaluronic acid; and 0.9% (w / v) of normal saline. The optimal formula of the digestive juice comprises the following components in concentration ratio: 0.1% (w / v) of type I collagenase; 0.125% (w / v) of EDTA; 0.25% (w / v) of trypsin; 0.02% (w / v) of DNA enzyme; 0.1% (w / v) of hyaluronic acid; and 0.9% (w / v) of normal saline. The digestive juice disclosed by the invention can be used for effectively separating pathogenic bacteria in a mucous specimen in a clinical microbiological test room; the activity of pathogenic bacteria in a mucous specimen in a clinical microbiological test room is effectively protected; and the convenience of application of digestive juice in a mucous specimen in a clinical microbiological test room is also taken into consideration.

Description

Technical Field

[0001] The invention belongs to the technical field of medical reagents, and in particular relates to a digestive fluid formula for sputum specimens in a clinical microbiology laboratory, a preparation method and an application thereof. Background Art

[0002] Microorganisms are widely distributed in nature. They are a class of lower organisms with tiny bodies, numerous types and numbers. Among them, a class of microorganisms that can cause pathogenicity to humans and animals are called pathogenic microorganisms. In recent years, diseases caused by pathogenic microorganisms, such as bacterial pneumonia, fungal pneumonia, mycoplasma pneumonia, and new coronavirus infections, are highly contagious and can usually cause a global pandemic. Therefore, in the detection of pathogenic microorganisms, it is necessary to be accurate, fast, efficient and simple. Effective separation of pathogenic microorganisms can greatly improve the timeliness and accuracy of test results, and play an important role in clinical medical diagnosis and treatment.

[0003] The current problems are as follows:

[0004] In the patient's lungs and trachea, due to the continuous inflammation, the lymph follicles in the posterior pharyngeal wall and the root of the tongue continue to proliferate. After the proliferation, they continue to secrete substances, which are coughed into the throat to form sputum. Sputum is specifically composed of the following components: airway secretions (mainly mucus), necrotic and detached mucosa, dead white blood cells, necrotic tissue, and a certain number of pathogens. Sputum samples are commonly used clinical microbiology test samples and often contain a variety of microbial components. However, when some patients are taking specimens for testing, viscous sputum specimens are sampled, especially critically ill patients in the respiratory department and ICU patients.

[0005] Because the composition of sputum is complex, and some clinical pathogens have high requirements for culture nutrition and grow slowly. If the sputum specimen contains a sticky part during culture, the pathogens are not easy to dissociate from the mucus part and it is difficult for them to absorb nutrients and oxygen in the culture medium. This leads to slow growth or death, which is not conducive to the detection of difficult and rare bacteria. Therefore, for clinical sputum samples that may contain pathogens, they need to be homogenized before culture to liquefy the specimens and separate single pathogen colonies.

[0006] When the viscous sputum specimens are digested by conventional reagents such as sulfuric acid, sodium chloride, sodium hydroxide, N-acetyl-L-cysteine ​​(NALC), benzalkonium bromide, oxalic acid, pancreatic enzymes, dithiothreitol (DTT), etc., it is difficult to completely separate the single pathogenic colonies contained in the viscous part of the sputum. Once the pathogens cannot be separated, it directly affects the subsequent culture, identification and analysis. It is easy to cause false negative results, miss positive patients, and reduce the quality of hospital medical care. It can be seen that the optimization of the digestion and processing of viscous sputum specimens in the clinical microbiology laboratory of the laboratory department is a problem that needs to be solved urgently by the majority of medical workers.

[0007] The current solutions and technical problems are as follows:

[0008] 1. Use sterile water, N-acetyl-L-cysteine ​​(NALC), benzalkonium bromide and dithiothreitol (DTT) and other solutions for dilution, and then inoculate and culture the specimen. Since the sterile aqueous solution is not an isotonic solution, it can neither protect the activity of pathogens in the mucus well nor effectively destroy the structure of fibrin in the mucus; although solutions such as N-acetyl-L-cysteine ​​(NALC), benzalkonium bromide and dithiothreitol (DTT) can dissociate fibrin in the mucus, they cannot protect the activity of pathogens in the mucus well; all of these lead to the inability to separate active pathogens from the mucus to the greatest extent. Therefore, it affects the isolation and culture of clinical pathogens. It also makes subsequent identification and analysis unsustainable.

[0009] 2. Use sodium chloride solution for dilution, and then inoculate and culture the specimen. Since sodium chloride solution is an isotonic solution, although it can protect the activity of pathogens in the sputum, it cannot effectively destroy the structure of fibrin in the sputum, resulting in the inability to separate the pathogens in the sputum to the greatest extent. It is not conducive to the separation and cultivation of clinical pathogens. It also makes subsequent identification and analysis impossible.

[0010] 3. Use sulfuric acid, oxalic acid and potassium hydroxide solution for dilution and digestion, and then inoculate and culture the specimen. Since the pH value of sulfuric acid, oxalic acid and potassium hydroxide solution is strongly acidic / alkaline, although it can destroy the structure of substances such as fibrin in sputum, it also damages the activity of pathogens, and the survival rate of isolated pathogens decreases, which is not conducive to the pure culture of clinical pathogens, reduces the positive rate, is easy to miss the diagnosis, and makes subsequent identification and analysis unsustainable.

[0011] 4. Use pancreatic enzyme solution for dilution and digestion, and then inoculate and culture the specimen. Since the digestive corrosivity of pancreatic enzyme solution is best at a temperature of 37 degrees Celsius, the temperature requirement limits the convenient application of pancreatic enzyme solution to digest sputum specimens. Moreover, multiple warm baths of pancreatic enzyme solution lead to a decrease in the affinity and titer of the enzyme in the pancreatic enzyme solution. In addition, although the high corrosiveness of pancreatic enzyme solution can destroy the fiber structure in the sputum, it also damages the activity of pathogens, and the survival rate of isolated pathogens decreases, affecting the pure culture of clinical pathogens. It also makes subsequent identification and analysis unsustainable.

[0012] At present, the patents related to digestive fluid of sputum specimens in clinical microbiology laboratories mainly focus on whether there are pathogens in the sputum specimens, but do not emphasize the degree of separation and the ease of use. Therefore, there is insufficient technical exploration of the improvement of the quality of clinical medical testing technology. Summary of the invention

[0013] In order to solve the problems existing in the prior art, the present invention provides a digestive juice formula, a preparation method and an application of a sputum sample in a clinical microbiology laboratory.

[0014] The technical solution adopted by the present invention to solve this problem is:

[0015] A digestive solution for sputum specimens in a clinical microbiology laboratory, comprising the following components and concentration ratios:

[0016]

[0017] In the above technical solution, the optimal formula of digestive juice for sputum specimens in clinical microbiology laboratories includes the following components and concentration ratios:

[0018]

[0019] The second invention object of the present invention is to provide a method for preparing digestive juice of sputum specimens in a clinical microbiology laboratory, comprising the following steps:

[0020] S101, prepare 15% normal saline, 12.5% ​​type I collagenase, 12.5% ​​EDTA, 15.625% trypsin, 20% DNase and 20% hyaluronic acid as the stock solution;

[0021] S102, using the above stock solution to prepare a 10-fold concentrated storage solution according to the following proportions:

[0022]

[0023] S103, after the storage solution is mixed evenly, it is sterilized by filtering through a 0.22um filter membrane, and then packaged and stored in a -20 degree refrigerator;

[0024] S104. Before use, restore the -20°C digestion working solution to room temperature or warm it to 37°C. Take a specific volume of storage solution and mix it with 9 times the volume of distilled water to dilute it into the working solution.

[0025] The third inventive object of the present invention is to provide an application of a digestive fluid for sputum specimens in a clinical microbiology laboratory. The digestive fluid described above is used for digestion and dissociation of sputum specimens in a clinical microbiology laboratory, effective separation of active pathogens in sputum specimens, effective protection of the activity of pathogens in sputum specimens, or as a digestive fluid for routine bacterial culture of sputum specimens.

[0026] The fourth inventive object of the present invention is to provide a digestive fluid preparation for sputum specimens in a clinical microbiology laboratory, comprising the digestive fluid described above and optionally one or more pharmaceutically acceptable carriers.

[0027] In the above technical solution, the digestive fluid preparation is a liquid substance including digestive fluid in the form of liquid, paste, spray, concentrated tablet or pill.

[0028] A fifth object of the present invention is to provide a kit comprising the digestive fluid described above.

[0029] The advantages and positive effects of the present invention are:

[0030] 1. The digestive solution of the present invention can effectively separate pathogens in sputum specimens of clinical microbiology laboratories; effectively protect the activity of pathogens in sputum specimens of clinical microbiology laboratories; and also take into account the convenience of using the digestive solution in sputum specimens of clinical microbiology laboratories.

[0031] 2. The present invention improves the efficiency, process and quality of pathogen isolation in sputum specimens in clinical microbiology laboratories, avoids missed diagnosis, improves the patient's examination experience, and improves the medical quality of medical institutions and the public's satisfaction with the services of medical institutions.

[0032] 3. At present, there is no scientific observation and experiment on the digestion technology of sticky phlegm at home and abroad, and no research and development of digestive fluid has been carried out to complete the optimization of sticky phlegm digestion. The technical solution of the present invention fills the technical gap in the industry at home and abroad. As a new commercial or self-configured sticky phlegm digestive fluid sample or commodity, the technical solution of the present invention is expected to obtain considerable benefits and commercial value after transformation. BRIEF DESCRIPTION OF THE DRAWINGS

[0033] The technical solution of the present invention will be further described in detail below in conjunction with the accompanying drawings and embodiments, but it should be understood that these drawings are designed only for explanation purposes and are not intended to limit the scope of the present invention. In addition, unless otherwise specified, these drawings are intended only to conceptually illustrate the structural configurations described herein and are not necessarily drawn to scale.

[0034] Figure 1 : Comparison of properties of digestive juice formula after digestion of sputum specimens in clinical microbiology laboratory;

[0035] Figure 2 :Comparison of colonies after inoculation and culture of sputum specimens in clinical microbiology laboratory;

[0036] Figure 3 :Statistical comparison of colony counts after inoculation and culture of sputum specimens in clinical microbiology laboratories;

[0037] Figure 4 : Comparison of properties of digestive solution formula 3, currently available digestive solution, and blank control after digestion of sputum specimens from clinical microbiology laboratory;

[0038] Figure 5 : Comparison of colonies after inoculation and culture of digestive fluid formula 3, currently available digestive fluid, and blank control digestive sputum specimens from clinical microbiology laboratory;

[0039] Figure 6 : Statistical comparison of the number of colonies in the sputum specimens of the clinical microbiology laboratory after inoculation and culture of the digestive fluid formula 3, the currently available digestive fluid, and the blank control digestive fluid. DETAILED DESCRIPTION

[0040] In order to make the above-mentioned objects, features and advantages of the present invention more obvious and easy to understand, the specific implementation methods of the present invention are described in detail below in conjunction with the accompanying drawings. In the following description, many specific details are set forth to facilitate a full understanding of the present invention. However, the present invention can be implemented in many other ways different from those described herein, and those skilled in the art can make similar improvements without violating the connotation of the present invention, so the present invention is not limited to the specific implementation disclosed below. In addition, any single technical feature described or implied in the embodiments mentioned herein, or any single technical feature displayed or implied in the accompanying drawings, can still be combined or deleted arbitrarily between these technical features (or their equivalents), so as to obtain more other embodiments of the present invention that may not be directly mentioned in this article.

[0041] The present invention is further described in detail below in conjunction with specific embodiments. It should be understood that the specific embodiments described herein are only used to explain the present invention and are not used to limit the present invention.

[0042] Under the condition of high secretion of airway mucus, the mucins that make up the mucus secreted in sputum are mainly mucin 5AC (MUC5AC) secreted by airway goblet epithelial cells; long-term and repeated airway inflammation can often cause bronchial epithelial goblet cell metaplasia and secrete a large amount of MUC5AC. MUC5AC is a highly O-glycosylated macromolecular glycoprotein, composed of a protein backbone rich in serine, threonine, and proline and oligosaccharide branched side chains, with a complex molecular structure.

[0043] DNA enzyme, sterile water or physiological saline can regulate the viscous mucus part of sputum, mainly by affecting the formation of DNA mucin cross-links to make it easy to dissociate. Type I collagenase, EDTA, trypsin, and hyaluronic acid can dissolve and digest the viscous mucus part of sputum, mainly by reducing the viscosity of sputum to make it easy to dissociate. At the same time, hyaluronic acid plays a role in water retention, maintaining the extracellular space of pathogenic microorganisms, osmotic pressure, lubrication, and preventing pathogenic microorganisms from being damaged by excessive digestion during sputum digestion. This is because hyaluronic acid molecules contain a large number of carboxyl and hydroxyl groups, which form hydrogen bonds between molecules and molecules in aqueous solution, which makes it have a strong water retention effect and can bind more than 400 times its own water; at lower concentrations, due to the tertiary network structure formed by its intermolecular action, its aqueous solution does not show viscoelasticity, and also has the function of protecting the outer membrane of pathogenic microorganisms. Therefore, we designed these digestive solutions for formula combination and used clinical microbial mucous sputum specimens for experimental verification. It has been verified through experiments that the digestive fluid including the following components and concentration ratios can effectively separate the pathogens in the sputum specimens of the clinical microbiology laboratory; and effectively protect the activity of the pathogens in the sputum specimens of the clinical microbiology laboratory. The formula is as follows:

[0044]

[0045] Furthermore, the best combination of the above formula is as follows:

[0046]

[0047] The experimental verification process is described in detail below with reference to the accompanying drawings.

[0048] 1. Configuration of digestive juice:

[0049] Prepare 15% saline, 12.5% ​​type I collagenase, 12.5% ​​EDTA, 15.625% trypsin, 20% DNA enzyme and 20% hyaluronic acid as the stock solution, specifically:

[0050] Preparation of 0.5-15% physiological saline solution: add 5-150 g of common salt to every 1000 mL of distilled water. Preferably, add 150 g of common salt to every 1000 mL of distilled water and shake gently until completely dissolved.

[0051] 0.25-15% type I collagenase preparation: 250-15000 mg of powdered type I collagenase can be dissolved in 100 mL of distilled water. Preferably, add 12500 mg of powdered type I collagenase to every 100 mL of distilled water and shake gently until the collagenase is completely dissolved. Do not vortex to mix, add distilled water to make up to 100 ml, and wear gloves during the preparation process.

[0052] 0.25~15%EDTA configuration: 250~15000mg of powdered Na 2 EDTA is dissolved in 100 mL of distilled water. Preferably, 12500 mg of powdered Na 2 Gently shake EDTA until completely dissolved.

[0053] 0.5625-25.625% Trypsin Preparation: Dissolve 562.5-25625 mg of powdered trypsin in distilled water. Preferably, add 15625 mg of powdered trypsin to every 100 mL of distilled water. Shake gently until the protease is completely dissolved. Do not vortex to mix. Add distilled water to make up to 100 mL. Wear gloves during preparation.

[0054] 1-30% DNA enzyme preparation: 1000-30000 mg of powdered DNA enzyme is dissolved in 100 mL of distilled water. Preferably, 20000 mg of powdered DNA enzyme is added to each 100 mL of distilled water. Shake gently until the DNA enzyme is completely dissolved. Do not vortex to mix. Add distilled water to make up to 100 ml. Wear gloves during preparation.

[0055] 1-30% hyaluronic acid preparation: 1000-30000 mg of hyaluronic acid is dissolved in 100 mL of distilled water. Preferably, 20000 mg of hyaluronic acid is added to every 100 mL of distilled water, and the mixture is gently shaken until completely dissolved, and then the volume is fixed to 100 ml with distilled water.

[0056] 2. Specific examples of experiments:

[0057] Use the above stock solution to dilute 10 times as the working solution, and dissolve 1.25% type I collagenase, 1.25% EDTA, 1.5625% trypsin, 2% DNA enzyme and 2% hyaluronic acid in 1.5% saline to prepare digestion solution. Wear gloves during the preparation process. The above reagents are sterilized by filtering with a 0.22um filter membrane and then packaged and stored at -20℃ for use. The specific configuration method is shown in the following table (expressed in volume percentage (%), and the concentration deviation is controlled to ±1%).

[0058] Table 1: A table of the number of portions of digestive juice prepared for sputum specimens in a clinical microbiology laboratory

[0059]

[0060] Return the -20℃ digestion working solution to room temperature. If conditions permit, warm it to 37℃.

[0061] A clinical sputum specimen was collected and divided into 5 equal parts. The digestive solutions of each formula were added. After homogenization, the samples were vortexed for 20 seconds and allowed to stand at 30-37°C for 15-20 minutes. Then, the subsequent operations were performed. Each sample was operated in the same way and 3 blood plates were inoculated. According to experimental comparison and statistics, it was finally concluded that the digestive solution formula 3 had the best effect, and its optimal concentration was: 0.1% type I collagenase, 0.125% EDTA, 0.25% trypsin, 0.02% DNA enzyme, 0.1% hyaluronic acid and 0.9% sodium chloride.

[0062] 3. The experimental verification process is as follows:

[0063] S1. Deep coughed sputum samples are easy to collect in the early morning. Rinse your mouth thoroughly with cold boiled water before collection to prevent the normal oral flora from contaminating the sample.

[0064] S2. Samples should be sent for testing immediately after collection.

[0065] S3. When using in a biosafety cabinet, dilute the digestion solution to the working solution and return to room temperature or warm it to 37°C for later use.

[0066] S4. Add an equal amount of sputum digestion solution to the specimen for homogenization, vortex for 20 seconds, and let it stand at 30-37℃ for 15-20 minutes.

[0067] S5. Observe the degree of dissociation of the specimen. If the degree of dissociation is not good, repeat the operation and add equal doses of digestion solution until digestion and dissociation are observed with the naked eye, and then proceed to the next step of inoculation experiment operation.

[0068] S6. Place the digested sputum specimen and the blood agar plate to be inoculated in a biosafety cabinet. The operator holds a metal inoculation loop. After the inoculation loop is sterilized and cooled, the loop is immersed in the digested sputum specimen. The specimen (about 10 μl) is dipped and inoculated into the spare blood agar plate using the three-zone streak method. The plate is inverted at 5% CO. 2 Incubate in an incubator at 80% humidity and (36±1)℃. Observe the growth of the colonies after 24 hours.

[0069] It should be noted that according to the viscosity of the sputum specimens in the clinical microbiology laboratory, the dissociation degree of the viscous part of the sputum specimens in the clinical microbiology laboratory is adjusted by increasing the content of digestive juice, the digestion time, and the strength and number of external mixing of the specimens.

[0070] 4. The experimental results are as follows:

[0071] like Figure 1-3 As shown, Figure 1 This is a comparison chart of the properties of the digestive juice formula after digesting the sticky sputum specimens in the clinical microbiology laboratory; Figure 2 This is a comparison chart of colonies after inoculation and culture of sputum specimens in the clinical microbiology laboratory; Figure 3 It is a statistical comparison chart of the number of colonies after inoculation and cultivation of sputum specimens in clinical microbiology laboratories. The results show that the number of colonies obtained by treating sputum specimens with the digestive solution of the present invention is significantly more than that of existing digestive solutions, proving that the digestive solution of the present invention can effectively separate and protect pathogens. Further, the present invention inoculates the specimen with a culture medium, mixes it with Sabau agar cooled to about 45°C, pours it into an inoculation plate, and cultures it in a 35°C incubator for 48 hours, cultures it continuously for 5 days, and observes the results day by day. If suspicious fungal and yeast-like colonies are found, transfer to Sabau bacteria slant. It can be seen from the comparison that digestive solution formula 3 has the best effect, and is significantly better than the effects of other groups of formulas. Therefore, the optimal concentration of the above digestive solution formula is determined to be: 0.1% type I collagenase, 0.125% EDTA, 0.25% trypsin, 0.02% DNA enzyme, 0.1% hyaluronic acid and 0.9% sodium chloride.

[0072] In order to further verify the effect of digestive fluid formula 3, we compared it with the currently available digestive fluid (product of Qingdao Jieshikang Biotechnology Co., Ltd.; name: sputum digestive fluid (Sputasol method) product number: T1430; specification 100ml; main components: dithiothreitol, sodium chloride, disodium hydrogen phosphate, potassium dihydrogen phosphate) and a blank control (no digestive fluid treatment process), and the experiment was repeated three times.

[0073] like Figure 4-6 As shown, Figure 4The figure is a comparison of the properties of digestive solution formula 3, currently available digestive solution, and blank control after digestion of sputum specimens from a clinical microbiology laboratory; Figure 5 It is a comparison chart of colonies after inoculation and culture of digestive solution formula 3, currently available digestive solution, and blank control digestive sputum specimens from clinical microbiology laboratory; Figure 6 It is a statistical comparison chart of the colony counts of digestive solution formula 3, currently available digestive solutions, and blank control digestive sputum specimens after inoculation and culture in clinical microbiology laboratories.

[0074] By comparison, it can be seen that digestive juice formula 3 has the best effect and is significantly better than the effects of other groups of formulas. Therefore, the above digestive juice formula 3 is determined to be the best.

[0075] like Figure 1-6 As shown, the digestive solution prepared according to digestive solution formula 1-5 can effectively digest, dissociate and separate the pathogens in the sputum specimens of the clinical microbiology laboratory; can effectively protect the activity of the pathogens in the sputum specimens of the clinical microbiology laboratory; and can be used as a digestive solution for conventional bacterial culture of sputum specimens.

[0076] Furthermore, in order to facilitate the storage, transportation and use of digestive solution formula 3, we hereby explain the preparation method of digestive solution formula 3 storage solution: first prepare a 10-fold concentrated storage solution of digestive solution formula 3: For example: it is expected to prepare XmL storage solution (X>4), and it is prepared according to the volume ratio of formula 3, that is: 0.08*XmL of 12.5% ​​type I collagenase, 0.10*XmL of 12.5% ​​EDTA, 0.16*XmL of 15.625% trypsin, 0.01*XmL of 20% DNA enzyme and 0.05*XmL of 20% hyaluronic acid; finally, add 0.6*XmL of 15% saline, make up to XmL, and control the concentration deviation to ±0.1%, that is, the storage solution, and store it in a -20 degree refrigerator for subsequent experiments.

[0077] Before use, take a certain volume of storage solution and mix it with 9 times the volume of distilled water, that is, dilute it 1:10 to become a working solution. The final content is: 0.1% type I collagenase, 0.125% EDTA, 0.25% trypsin, 0.02% DNA enzyme and 0.1% hyaluronic acid mixed with 0.9% sodium chloride digestive solution. This formula is the best combination formula, and the final concentration ratio is: 0.1% type I collagenase, 0.125% EDTA, 0.25% trypsin, 0.02% DNA enzyme, 0.1% hyaluronic acid and 0.9% sodium chloride; the concentration deviation is controlled to ±0.1%, and the content of each component is >0; further, in order to facilitate application, transportation and storage, the liquid of this composition can also be made into liquid, ointment, spray, concentrated tablet, pill and other dosage forms; the liquid of this composition can also be made into a kit, which can take into account the convenience of the application of digestive fluid in sputum specimens in clinical microbiology laboratories.

[0078] The above embodiments describe the present invention in detail, but the contents are only preferred embodiments of the present invention and cannot be considered to limit the scope of the present invention. All equivalent changes and improvements made within the scope of the present invention should still fall within the scope of the present invention.

Claims

1. A digestive fluid for sputum specimens in a clinical microbiology laboratory, characterized in that: The components and concentration ratios are as follows:

2. The digestive fluid for preparing sputum specimens in a clinical microbiology laboratory according to claim 1, characterized in that: The components and concentration ratios are as follows:

3. A method for preparing digestive juice of sputum specimens in clinical microbiology laboratories as claimed in claim 1 or 2, characterized in that: The steps include: S101, prepare 15% normal saline, 12.5% ​​type I collagenase, 12.5% ​​EDTA, 15.625% trypsin, 20% DNase and 20% hyaluronic acid as the stock solution; S102, using the above stock solution to prepare a 10-fold concentrated storage solution according to the following proportions: S103, after the storage solution is mixed evenly, it is sterilized by filtering through a 0.22um filter membrane, and then packaged and stored in a -20 degree refrigerator; S104. Before use, restore the -20°C digestion working solution to room temperature or warm it to 37°C. Take a specific volume of storage solution and mix it with 9 times the volume of distilled water to dilute it into the working solution.

4. An application of digestive juice for sputum specimens in a clinical microbiology laboratory, characterized in that: The digestive solution of the clinical microbiology laboratory sputum specimen as claimed in claim 1 or 2 or the digestive solution prepared by the preparation method as claimed in claim 3 is used as the digestive solution for digestion and dissociation of the clinical microbiology laboratory sputum specimen.

5. An application of digestive juice for sputum specimens in a clinical microbiology laboratory, characterized in that: The digestive fluid of the clinical microbiology laboratory sputum specimen as described in claim 1 or 2 or the digestive fluid prepared by the preparation method as described in claim 3 is used as a digestive fluid for effectively separating active pathogens in the clinical microbiology laboratory sputum specimen and effectively protecting the activity of pathogens in the sputum specimen.

6. An application of digestive juice for sputum specimens in a clinical microbiology laboratory, characterized in that: The digestive fluid of the clinical microbiology laboratory sputum specimen as claimed in claim 1 or 2 or the digestive fluid prepared by the preparation method as claimed in claim 3 is used as the digestive fluid for routine bacterial culture of the clinical microbiology laboratory sputum specimen.

7. A digestive fluid preparation for sputum specimens in a clinical microbiology laboratory, characterized in that: The invention comprises the digestive juice of the sputum specimen of the clinical microbiology laboratory as claimed in claim 1 or 2 or the digestive juice prepared by the preparation method as claimed in claim 3, and optionally one or more pharmaceutically acceptable carriers.

8. The digestive fluid preparation for sputum specimens in clinical microbiology laboratories according to claim 7, characterized in that: The digestive fluid preparation is a liquid, paste, spray, concentrated tablet or pill made from the liquid substance of the digestive fluid.

9. A kit, characterized in that: It contains the digestive juice of the sputum specimen of the clinical microbiology laboratory as claimed in claim 1 or 2 or the digestive juice prepared by the preparation method as claimed in claim 3.

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