CN109001455B - Alveolar lavage fluid sample treatment fluid and treatment detection method - Google Patents
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Abstract
The invention discloses an alveolar lavage fluid sample treatment fluid and a treatment and detection method, wherein the treatment fluid comprises inorganic salt and a surfactant, and the treatment fluid can reduce the interference of other substances in the alveolar lavage fluid on the detection of fungi (1-3) -beta-D-glucan. The invention processes the alveolar lavage fluid sample, is used for detecting the content of the fungus (1-3) -beta-D-glucan in the alveolar lavage fluid of an invasive pulmonary fungal infection patient in vitro, has higher detection sensitivity compared with the traditional serum detection method, and can keep higher specificity and accuracy.
Description
Technical Field
The invention belongs to the field of in vitro detection of fungi (1-3) -beta-D-glucan, and particularly relates to an alveolar lavage fluid sample treatment fluid and a treatment and detection method.
Background
The invention is used for auxiliary diagnosis of invasive deep fungal infection in clinic. With the continuous development of clinical technologies such as the use of a large amount of corticosteroids, immunosuppressants, various broad-spectrum antibiotics, organ transplantation and the like, the probability of invasive fungal infection is generally increased for some patients with low immunity. The (1-3) -beta-D glucan is a cell wall component of most medically important fungi (including candida and aspergillus), and can be released continuously after phagocytosis of the fungi by human phagocytes, so that the concentration of the (1-3) -beta-D glucan in human body fluids such as blood, alveolar lavage fluid, cerebrospinal fluid and the like is increased, and therefore, the detection of the (1-3) -beta-D glucan is a key index. At present, most of G test samples are serum, and compared with the patients with invasive pulmonary fungal infection, only part of glucan enters blood and the content of glucan is far lower than that of alveolar lavage fluid, so that the detection rate is higher for the patients with invasive pulmonary fungal infection.
The G test alkali treatment method is adopted by hospitals due to high sensitivity, high specificity and short detection time of the detection of (1-3) -beta-D glucan. Before, a hospital mostly adopts a heating dilution method to pretreat a sample, but the method hardly enables the form (single-stranded form, double-stranded helix or triple-stranded helix) of (1-3) -beta-D glucan in the sample to be uniform, so that the detection specificity of the (1-3) -beta-D glucan is influenced, and foreign researchers research shows that the (1-3) -beta-D glucan can be converted into the triple-stranded helix after heat treatment, and the triple-stranded helix structure can be hydrolyzed into the single-stranded structure after alkali treatment. Therefore, the invention uses G test alkali treatment method for experimental study.
The constituents of alveolar lavage fluid include cells, lung surfactant (PS), inflammatory mediators, growth factors, various kinds of eggs, enzymes, lipids, and the like. The existence of certain interference factors in the alveolar lavage fluid sample components can seriously affect the determination result of (1-3) -beta-D glucan, and the specificity and the accuracy of the determination are reduced. The reaction interference factor refers to a reaction factor (false positive factor) which is not involved in a given biochemical reaction or a factor (false negative factor) which has an interference effect on the reaction at any stage of the reaction. The reaction sample, i.e., alveolar lavage fluid, contains such factors. For example, factor Xa shows a similar effect to the coagulase in the lysate and thus is a false positive factor. Alpha 1-antitrypsin has a strong interfering effect on the reaction and therefore acts as a false negative factor.
The function of the alkali metal hydroxide is to denature the reaction interference factor. Strong alkali breaks hydrogen bonds of proteins, forms salts with free amino groups and the like, and breaks and generates chemical bonds in the change process. The denatured interferents are still present in solution in a solubilized state. The dilution heating method only uses simple physical change to denature and precipitate the protein, and the denatured protein precipitate may cause interference on the turbidity of the reaction. KCl has inhibitory effect on adsorption of dextran to blood components. The addition of the alkali metal halide prevents dextran from being adsorbed to blood components, so that the G-factor cascade reaction is smoothly activated by dextran, the reaction system is stabilized, and the reproducibility is improved. However, the single use of the treatment solution used in the conventional serum detection is not enough to completely remove the interference factors in the sample, and the accuracy of the detection result is still to be improved, so that it is important to solve the interference problem in the detection of alveolar lavage fluid, and it avoids unnecessary clinical examination and unreasonable treatment in the later period of the patient.
Disclosure of Invention
In order to solve the problem of detection interference, the invention provides an alveolar lavage fluid sample treatment solution and a treatment detection method, wherein the G test alkali treatment method is utilized to eliminate or reduce interference factors influencing the detection of the fungus (1-3) -beta-D glucan in an alveolar sample, thereby achieving the purpose of improving the detection sensitivity and specificity of the fungus (1-3) -beta-D glucan.
The technical scheme adopted by the invention is as follows: a treatment fluid for an alveolar lavage fluid sample, comprising: the treatment solution A comprises alkali metal hydroxide and a surfactant, and the treatment solution B comprises alkali metal halide.
Preferably, the alkali metal hydroxide is one of hydrogen LiOH, NaOH, KOH.
Preferably, the alkali metal halide is KCl, MgCl2、CaCl2、SrCl2One kind of (1).
Preferably, the surfactant is one of the Triton series or the Tween series, preferably Triton X-100 or Tween 20.
Preferably, the treating fluid A specifically comprises KCl0.05-0.5M, Triton X-1000.01-1% or Tween 200.50-1.50%, and the treating fluid B specifically comprises KOH 0.05-0.5M.
Preferably, treatment fluid A comprises KCl0.5M, Triton X-1000.10%, and treatment fluid B comprises KOH 0.25M.
A method for detecting fungi by using sample treatment liquid to pretreat alveolar lavage liquid comprises the following specific steps:
the method comprises the following steps: preparing a treatment solution A which comprises 0.05-0.5M of KCl and 0.05-0.5M of KOH and 0.05-0.5M of Triton X-1000.01-1%, and mixing the treatment solution A and the treatment solution B in equal volume to obtain a sample treatment solution;
step two: adding a sample treatment solution into the alveolar lavage fluid sample, uniformly mixing, and then incubating;
step three: adding a reaction main agent for detecting (1-3) -beta-D glucan into the incubated mixed solution, shaking and uniformly mixing, detecting and calculating the content of the (1-3) -beta-D glucan.
Preferably, the sample processing solution specifically comprises KCl0.5M, KOH0.25M, Triton X-1000.10%.
Preferably, the volume ratio of the alveolar lavage fluid sample to the sample treatment fluid in step two is 1:3-5, preferably 1: 4.
Preferably, the incubation temperature in the second step is 37 ℃ and the incubation time is 10-15 min.
Preferably, the reaction main agent adopted in the third step comprises factor G, a coagulase source and a coagulating protein source, and the volume ratio of the alveolar lavage fluid to the reaction main agent is 1:15-25, preferably 1: 20.
The invention has the advantages and positive effects that:
the scheme 1 is used for detecting the alveolar lavage fluid, for patients with invasive pulmonary fungal infection, the content of (1-3) -beta-D glucan in serum is far lower than that of alveolar lavage fluid, the detection rate of the alveolar lavage fluid is higher, and the patients with invasive pulmonary fungal infection can be judged earlier;
2, compared with a serum sample, the alveolar lavage fluid sample contains more interference factors, and the sample treatment fluid related to the scheme can effectively remove the interference factors, so that the detection result is more accurate, and the specificity and the accuracy of the determination are improved;
3, the surfactant with low concentration can help to decompose protease, reduce the influence of interference factors of alveolar lavage fluid on detection, and improve the detection specificity by matching with alkali metal hydroxide and alkali metal halide;
4 this detection scheme easy operation, detection rate is high, can obtain the testing result fast.
Detailed Description
The following description is made of embodiments of the present invention.
The invention relates to a sample treatment solution for alveolar lavage fluid, which at least comprises the following components: alkali metal hydroxides, alkali metal halides, and surfactants. The alkali metal hydroxide includes hydroxides of alkali metals such as lithium, sodium, and potassium, and specifically includes lithium hydroxide (LiOH), sodium hydroxide (NaOH), and potassium hydroxide (KOH). The treatment solution is usually an aqueous solution of one or more alkali metal hydroxides, and NaOH or KOH is used for higher detection effect and economic benefit through experimental comparison. The alkali metal halide includes potassium chloride, magnesium chloride, calcium chloride, strontium chloride, etc., but potassium chloride is preferable as the treatment liquid because ions such as magnesium and calcium are easily bonded to hydroxide radicals to generate precipitates. The non-ionic surfactant is selected from Triton series or Tween series.
The treatment liquid of the present invention may be stored as two separate solutions, and these solutions may be mixed together at the time of use. If the alkali metal oxide and the alkali metal hydroxide or the nonionic surfactant are mixed together and stored for a long period of time, there is an inconvenience such as precipitation, and therefore, they are stored separately until immediately before pretreatment. Therefore, the alveolar lavage fluid sample treatment fluid is divided into a treatment fluid A containing an alkali metal halide and a nonionic surfactant, and a treatment fluid B containing an alkali metal hydroxide.
A treatment solution for alveolar lavage fluid sample comprises a treatment solution A and a treatment solution B, wherein the treatment solution A comprises KCl and a surfactant, wherein the treatment solution comprises KCl0.05-0.5M and the surfactant 0.01-1%; the treating solution B is KOH solution, specifically KOH 0.05-0.5M. The surfactant may be Triton X-100 or Tween 20.
A large number of comparative tests show that the treatment effect is best when the treatment liquid A is KCl0.5M and Triton X-1000.10% and the treatment liquid B is KOH 0.25M.
A method for detecting fungi by using sample treatment liquid to pretreat alveolar lavage liquid comprises the following specific steps:
the method comprises the following steps: preparing a treatment solution A which comprises KCl0.05-0.5M, Triton X-100/Tween 200.01-1%, preparing a treatment solution B which is KOH0.05-0.5M, and mixing the treatment solution A and the treatment solution B in equal volume to obtain a sample treatment solution;
step two: adding a sample treatment solution into the alveolar lavage fluid sample, adding the sample treatment solution into the alveolar lavage fluid sample according to the volume ratio of 1:3-5, uniformly mixing, and incubating at 37 ℃ for 10-15 min;
step three: adding a reaction main agent for detecting (1-3) -beta-D glucan into the incubated mixed solution, wherein the volume ratio of the alveolar lavage fluid to the reaction main agent is 1:15-25, shaking, mixing uniformly, detecting and calculating the content of the (1-3) -beta-D glucan.
When the volume ratio of the alveolar lavage fluid sample to the sample treatment fluid is 1:4 and the volume ratio of the alveolar lavage fluid sample to the reaction main agent is 1:20, the test efficiency is higher.
Wherein the preparation steps of the main reaction agent are as follows
1, separating and extracting a horseshoe crab blood cell lysate, performing aseptic operation, taking about 20g of blood cells, adding a blood cell lysate according to a proportion of 1-5, performing 200000rpm/min by using a high-speed homogenizer for 5-10 minutes of damaged cells, placing the damaged histiocyte sap into a refrigerator at minus 30 ℃ for rapid freezing and storing for 10 hours, then taking out the frozen histiocyte sap, slowly dissolving at room temperature, performing 200000rpm/min by using the high-speed homogenizer for 5-10 minutes of damaged cells after all the cells are dissolved, then placing the damaged histiocyte sap into the refrigerator at minus 30 ℃ for rapid freezing and storing for 10 hours again, and repeating the previous process for 2 times.
2, taking out the blood cell lysate, centrifuging at 3000rpm/min for 5-8 minutes, taking supernatant, and performing centrifugation according to the ratio of 1: adding chloroform at the ratio of 2, violently shaking for 10 minutes, transferring to a pyrogen-free centrifugal sedimentation tube, performing 10000rpm/min at the temperature of 4 ℃, separating for 5-10 minutes, and carefully taking out a supernatant for later use.
3 taking the supernatant of the extracted and separated horseshoe crab blood cells, adding the supernatant into 0.5M calcium chloride solution containing 1.0M magnesium chloride, subpackaging the mixture into a penicillin bottle or an ampoule bottle, and freeze-drying the mixture to obtain the main reaction agent.
The main reaction agent adopted in the scheme can also be directly adopted in a commercially available (1-3) -beta-D glucan serum detection kit, for example, the main reaction agent in a fungus (1-3) -beta-D glucan detection kit (a color development method) of Tianjin Xinuo biological medicine limited company is adopted.
The detection solution after mixing in the third step can be detected by a microorganism rapid dynamic detection system (MB-80A, MB-80M, MB-80S, MB-80X), a fungus/bacterium dynamic detector (MB-80A, MB-80X, MB-80M) or a full-automatic fungus/bacterium dynamic detector (IGL-800), and the specificity and accuracy of the detection scheme are verified by comparing the detection data with the culture result of the detected substance in the following examples.
For the treating fluid B, the concentration range of the KOH used as the sample treating fluid is 0.05-0.5M, in order to further determine the optimum concentration of the KOH, the treating fluid only contains the KOH, 100pg/mL of fungus standard substance is added into the alveolar sample, KOH with different concentrations is detected, the influence on the test recovery rate of the sample addition standard substance is detected, and the detection results are shown in Table 1.
TABLE 1
The test result shows that the glucan content can not be detected only by using sterile water to treat a sample, the recovery rate is gradually increased along with the increase of the KOH concentration, and the recovery rate is within 50-150% when the KOH concentration is 0.05-0.40M, which indicates that the detection result is significant. In combination with the culture result of the sample, the detection result is negative when the KOH concentration is 0.15-0.50M, so that the KOH concentration is more suitable for treating the alveolar lavage fluid sample when the KOH concentration is 0.15-0.40M. The data in Table 1 show that the recovery rate is not high when the treating solution contains KOH only, which indicates that there is still some interference, and the addition of alkali metal halide and nonionic surfactant to the treating solution can increase the recovery rate and reduce the interference. In the subsequent detection, the KOH concentration was fixed, and one of the intermediate concentrations was selected to be 0.25M, and the effects of the other components were analyzed.
Example 1
A sample treatment solution for alveolar lavage fluid comprises the following raw materials: treatment liquid A (KCl 0.12M, Triton X-1000.01%) and treatment liquid B (KOH 0.25M).
The method for detecting the fungi by using the sample treatment solution to pretreat the alveolar lavage solution comprises the following specific steps:
the method comprises the following steps: respectively preparing a treatment solution A and a treatment solution B, and mixing the treatment solution A and the treatment solution B in equal volume to obtain a sample treatment solution;
step two: adding 40 μ L of the sample treatment solution into 10 μ L of alveolar lavage solution sample, mixing, and incubating at 37 deg.C for 10 min;
step three: and adding 200 mu L of reaction main agent into the incubated mixed solution, shaking and uniformly mixing, detecting and calculating the content of the (1-3) -beta-D glucan.
Example 2
A sample treatment solution for alveolar lavage fluid comprises the following raw materials: treatment liquid A (KCl 0.12M, Triton X-1000.10%) and treatment liquid B (KOH 0.25M).
Pretreatment of alveolar lavage fluid with sample treatment fluid the procedure for fungal detection was as in example 1.
Example 3
A sample treatment solution for alveolar lavage fluid comprises the following raw materials: treatment liquid A (KCl 0.12M, Triton X-1000.50%) and treatment liquid B (KOH 0.25M).
Pretreatment of alveolar lavage fluid with sample treatment fluid the procedure for fungal detection was as in example 1.
Example 4
A sample treatment solution for alveolar lavage fluid comprises the following raw materials: treatment liquid A (KCl 0.12M, Triton X-1001.00%) and treatment liquid B (KOH 0.25M).
Pretreatment of alveolar lavage fluid with sample treatment fluid the procedure for fungal detection was as in example 1.
Example 5
A sample treatment solution for alveolar lavage fluid comprises the following raw materials: treatment liquid A (KCl 0.06M, Triton X-1000.10%) and treatment liquid B (KOH 0.25M).
Pretreatment of alveolar lavage fluid with sample treatment fluid the procedure for fungal detection was as in example 1.
Example 6
A sample treatment solution for alveolar lavage fluid comprises the following raw materials: treatment liquid A (KCl 0.12M, Triton X-1000.10%) and treatment liquid B (KOH 0.25M).
Pretreatment of alveolar lavage fluid with sample treatment fluid the procedure for fungal detection was as in example 1.
Example 7
A sample treatment solution for alveolar lavage fluid comprises the following raw materials: treatment liquid A (KCl 0.50M, Triton X-1000.10%) and treatment liquid B (KOH 0.25M).
Pretreatment of alveolar lavage fluid with sample treatment fluid the procedure for fungal detection was as in example 1.
Example 8
A sample treatment solution for alveolar lavage fluid comprises the following raw materials: treatment liquid A (KCl 0.50M, Triton X-1000.10%) and treatment liquid B (KOH 0.25M).
Pretreatment of alveolar lavage fluid with sample treatment fluid the procedure for fungal detection was as in example 1.
Example 9
A sample treatment solution for alveolar lavage fluid comprises the following raw materials: treatment liquid A (KCl 0.50M) and treatment liquid B (KOH 0.25M).
Pretreatment of alveolar lavage fluid with sample treatment fluid the procedure for fungal detection was as in example 1.
Example 10
A sample treatment solution for alveolar lavage fluid comprises the following raw materials: treatment solution A (Triton X-1000.10%), and treatment solution B (KOH 0.25M).
Pretreatment of alveolar lavage fluid with sample treatment fluid the procedure for fungal detection was as in example 1.
Example 11
A sample treatment solution for alveolar lavage fluid comprises the following raw materials: treatment liquid A (KCl 0.50M, Triton X-1000.05%) and treatment liquid B (KOH 0.25M).
Pretreatment of alveolar lavage fluid with sample treatment fluid the procedure for fungal detection was as in example 1.
Example 12 detection of alveolar lavage fluid by Tween20 as surfactant as treatment fluid
A sample treatment solution for alveolar lavage fluid comprises the following raw materials: treatment liquid A (KCl 0.50M, Tween 200.10%) and treatment liquid B (KOH 0.25M).
Pretreatment of alveolar lavage fluid with sample treatment fluid the procedure for fungal detection was as in example 1.
Example 13
A sample treatment solution for alveolar lavage fluid comprises the following raw materials: treatment liquid A (KCl 0.50M, Tween 200.50%) and treatment liquid B (KOH 0.25M).
Pretreatment of alveolar lavage fluid with sample treatment fluid the procedure for fungal detection was as in example 1.
Example 14
A sample treatment solution for alveolar lavage fluid comprises the following raw materials: treatment liquid A (KCl 0.50M, Tween 201.00%) and treatment liquid B (KOH 0.25M).
Pretreatment of alveolar lavage fluid with sample treatment fluid the procedure for fungal detection was as in example 1.
Example 15
A sample treatment solution for alveolar lavage fluid comprises the following raw materials: treatment liquid A (KCl 0.50M, Tween 201.50%) and treatment liquid B (KOH 0.25M).
Pretreatment of alveolar lavage fluid with sample treatment fluid the procedure for fungal detection was as in example 1.
Example 16
A sample treatment solution for alveolar lavage fluid comprises the following raw materials: treatment liquid A (KCl 0.50M, Tween 202.00%) and treatment liquid B (KOH 0.25M).
Pretreatment of alveolar lavage fluid with sample treatment fluid the procedure for fungal detection was as in example 1.
The results of the comparisons of examples 1-4 are shown in Table 2.
TABLE 2
In examples 1-4, the composition of treatment solution B was unchanged, the KCl concentration in treatment solution A was unchanged, the Triton X-100 concentration was gradually increased, and the Triton X-100 concentration was first decreased and then increased in the process of 0.1% -1.0% change, as is evident from the fact that the detection result was closest to the culture result and the greatest effect was obtained when the Triton X-100 concentration was 0.1% in example 2.
The results of the comparisons of examples 5 to 7 are shown in Table 3.
TABLE 3
In examples 5-7, the composition of treatment solution B was unchanged, the concentration of Triton X-100 in treatment solution A was unchanged, and the KCl concentration was gradually increased, and it can be seen that the results of several samples tested by the treatment solution formulation in example 7 were consistent with the culture results, and the greatest effect was obtained in removing the interference factors from alveolar lavage fluid, so that the best detection sensitivity was obtained when the KCl concentration in treatment solution A was selected to be 0.50M.
The results of the comparisons of examples 8 to 10 are shown in Table 4.
TABLE 4
In examples 8 to 10, the composition of the treatment solution B was not changed, and the treatment solution A was set to contain Triton X-100 and KCl, respectively, only KCl, only Triton X-100, and compared with the detection accuracy thereof, it was found from the test results that when KCl and Triton X-100 were simultaneously present in the treatment solution A in example 8, the detection results were more in agreement with the culture results, and the detection results of the treatment solution A containing only KCl or Triton X-100 had a higher error rate, thereby confirming that KCl and Triton X-100 were not indispensable for the alveolar lavage fluid as the detection sample. The solution only added with KCl is a low-molecular dispersion system, the main dispersion phase is molecules and ions, but the solution does not have the dispersion effect on other macromolecular substances in the alveolar lavage fluid; the surfactant is dissolved in water, the surface energy of the water can be remarkably reduced, the surface tension can be remarkably reduced during low concentration, the problem of aggregation of macromolecular substances in a sample can be solved, a large number of tests prove that the nonionic surfactant is added to form steric hindrance, a space system is stable, the aggregation is reduced, the glucan molecules are not prone to coagulation of other macromolecular impurities due to good space barriers, and therefore the detection result can be guaranteed to have higher accuracy only through the synergistic effect of the glucan molecules and the macromolecular impurities.
The results of comparing example 8 with example 11 are shown in Table 5.
TABLE 5
The test results of example 8 and example 11 respectively using 0.10% Triton X-100 and 0.05% Triton X-100 show that the accuracy of example 8 is significantly higher, some false positive results appear in example 11, and the interference of the treatment solution of the formula is reduced when the component A of the treatment solution of example 8 is 0.5M KCl and 0.05% Triton X-100, and the test result is more matched with the culture result.
The results of comparing example 8 with examples 12-16 are shown in Table 6.
TABLE 6
The data in the above table show that the detection result is consistent with that of example 8 (control) when the concentration of Tween20 is 0.50-1.50%, and matches with the culture result, which indicates that Tween20 as one of the components of the treatment solution can effectively remove the interference factor in the alveolar lavage fluid, and improve the sensitivity and specificity of the detection result of the alveolar lavage fluid.
Example 17 detection of sensitivity and specificity
The formulation of the treatment solution in example 8 was selected, and the sensitivity and specificity of detection using the test kit of the kino G test after treatment with the treatment solution were tested using the culture results as a control (100 BALF samples with culture information were selected for testing),
judging the detection result of the test kit of the XinuoG:
| negative of | ≤60pg/mL |
| Gray zone | 60-100pg/mL |
| Positive for | ≥100pg/mL |
The sensitivity and specificity calculation methods are as in table 7:
TABLE 7
Sensitivity: (A + C)/(A + C + E) × 100%
Specificity: (D + F)/(B + D + F) × 100%
The test results are shown in Table 8.
TABLE 8
Results analysis is shown in Table 9
TABLE 9
As can be seen from the data, the matching degree of the test result of the treating fluid component of the alveolar lavage fluid sample and the culture result in the embodiment 8 is high, and the sensitivity and the specificity of the data measured by using the Xinuo G test kit are both up to 90.0 percent and 92.5 percent, so that the detection accuracy of the kit on the alveolar lavage fluid sample is obviously improved.
Example 18 verification of detection accuracy after treatment by recovery of fungal (1-3) - β -D glucan standards
The treatment fluid formulation of example 8 was selected and tested for recovery of fungal (1-3) - β -D glucan standards added to alveolar lavage fluid samples for test significance:
the recovery rate calculation method comprises the following steps:
watch 10
As shown in Table 10, the recovery rate of 5 samples was determined experimentally and the results were all between 50% and 150%, indicating that the detection results obtained by treating alveolar lavage fluid with the treatment fluid have small interference and reliable data.
Although the embodiments of the present invention have been described in detail, the description is only for the preferred embodiments of the present invention and should not be construed as limiting the scope of the present invention. All equivalent changes and modifications made within the scope of the present invention shall fall within the scope of the present invention.
Claims (4)
1. A treatment fluid for an alveolar lavage fluid sample, comprising: detecting the content of (1-3) -beta-D glucan, wherein the content of the (1-3) -beta-D glucan comprises a treatment liquid A and a treatment liquid B, the treatment liquid A comprises an alkali metal halide and a surfactant, and the treatment liquid B comprises an alkali metal hydroxide;
the alkali metal hydroxide is one of LiOH, NaOH and KOH;
the alkali metal halide is KCl and MgCl2、CaCl2、SrCl2One of (1);
the surfactant is one of Triton series or Tween series.
2. The alveolar lavage fluid sample treatment fluid according to claim 1, wherein: the treating fluid A specifically comprises KCl0.05-0.5M, TritonX-1000.01-1% or Twen 200.50-1.50%, and the treating fluid B specifically comprises KOH 0.05-0.5M.
3. The alveolar lavage fluid sample treatment fluid according to claim 2, wherein: the treatment liquid A comprises KCl0.5M and TritonX-1000.10%, and the treatment liquid B comprises KOH0.25M.
4. The alveolar lavage fluid sample treatment fluid according to claim 1, wherein: the surfactant is TritonX-100 or Tween 20.
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| JP6432083B2 (en) * | 2012-12-28 | 2018-12-05 | 生化学工業株式会社 | Pretreatment agent and method for antithrombin III subjected to limulus test |
| CN204188621U (en) * | 2014-07-11 | 2015-03-04 | 厦门市鲎试剂实验厂有限公司 | A kind of dynamic color method detecting limulus kit for glucan in fungus |
| MA41020A (en) * | 2014-11-25 | 2017-10-03 | Evelo Biosciences Inc | PROBIOTIC AND PREBIOTIC COMPOSITIONS, AND THEIR METHODS OF USE FOR MODULATION OF THE MICROBIOME |
| CN107466319B (en) * | 2017-06-28 | 2020-08-14 | 丹娜(天津)生物科技股份有限公司 | GM hybridoma cell, monoclonal antibody, kit, preparation method and application thereof |
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