CN109001455A - A kind of bronchoalveolar lavage fluid sample treatment solution and processing detection method - Google Patents
A kind of bronchoalveolar lavage fluid sample treatment solution and processing detection method Download PDFInfo
- Publication number
- CN109001455A CN109001455A CN201810622529.5A CN201810622529A CN109001455A CN 109001455 A CN109001455 A CN 109001455A CN 201810622529 A CN201810622529 A CN 201810622529A CN 109001455 A CN109001455 A CN 109001455A
- Authority
- CN
- China
- Prior art keywords
- bronchoalveolar lavage
- fluid
- lavage fluid
- treatment solution
- sample
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Granted
Links
- 239000012530 fluid Substances 0.000 title claims abstract description 176
- 238000001514 detection method Methods 0.000 title claims abstract description 54
- 238000012545 processing Methods 0.000 title abstract description 9
- 238000000034 method Methods 0.000 claims abstract description 38
- 241000233866 Fungi Species 0.000 claims abstract description 36
- 239000004094 surface-active agent Substances 0.000 claims abstract description 11
- KWYUFKZDYYNOTN-UHFFFAOYSA-M Potassium hydroxide Chemical compound [OH-].[K+] KWYUFKZDYYNOTN-UHFFFAOYSA-M 0.000 claims description 116
- 239000001103 potassium chloride Substances 0.000 claims description 42
- 239000013504 Triton X-100 Substances 0.000 claims description 23
- 229920004890 Triton X-100 Polymers 0.000 claims description 23
- 238000006243 chemical reaction Methods 0.000 claims description 22
- 239000003795 chemical substances by application Substances 0.000 claims description 17
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 claims description 15
- GPRLSGONYQIRFK-MNYXATJNSA-N triton Chemical class [3H+] GPRLSGONYQIRFK-MNYXATJNSA-N 0.000 claims description 15
- 108010001062 polysaccharide-K Proteins 0.000 claims description 14
- 229920001213 Polysorbate 20 Polymers 0.000 claims description 13
- 239000003513 alkali Substances 0.000 claims description 13
- 239000000256 polyoxyethylene sorbitan monolaurate Substances 0.000 claims description 13
- 235000010486 polyoxyethylene sorbitan monolaurate Nutrition 0.000 claims description 13
- 150000008044 alkali metal hydroxides Chemical class 0.000 claims description 11
- 229920002307 Dextran Polymers 0.000 claims description 10
- WMFOQBRAJBCJND-UHFFFAOYSA-M Lithium hydroxide Chemical compound [Li+].[OH-] WMFOQBRAJBCJND-UHFFFAOYSA-M 0.000 claims description 9
- 150000004820 halides Chemical class 0.000 claims description 9
- TWRXJAOTZQYOKJ-UHFFFAOYSA-L Magnesium chloride Chemical compound [Mg+2].[Cl-].[Cl-] TWRXJAOTZQYOKJ-UHFFFAOYSA-L 0.000 claims description 8
- 238000011534 incubation Methods 0.000 claims description 8
- 238000002156 mixing Methods 0.000 claims description 8
- 230000009514 concussion Effects 0.000 claims description 5
- UXVMQQNJUSDDNG-UHFFFAOYSA-L Calcium chloride Chemical compound [Cl-].[Cl-].[Ca+2] UXVMQQNJUSDDNG-UHFFFAOYSA-L 0.000 claims description 4
- 239000001110 calcium chloride Substances 0.000 claims description 4
- 229910001628 calcium chloride Inorganic materials 0.000 claims description 4
- 229910001629 magnesium chloride Inorganic materials 0.000 claims description 4
- 229920000136 polysorbate Polymers 0.000 claims description 3
- 108090000623 proteins and genes Proteins 0.000 claims description 3
- 102000004169 proteins and genes Human genes 0.000 claims description 3
- 229910001631 strontium chloride Inorganic materials 0.000 claims description 3
- 108010065152 Coagulase Proteins 0.000 claims description 2
- 230000035945 sensitivity Effects 0.000 abstract description 10
- 210000002966 serum Anatomy 0.000 abstract description 6
- 208000013606 Fungal Lung disease Diseases 0.000 abstract description 4
- 239000000126 substance Substances 0.000 abstract description 4
- 229920000018 Callose Polymers 0.000 abstract description 3
- 150000003839 salts Chemical class 0.000 abstract description 2
- 238000000338 in vitro Methods 0.000 abstract 1
- 230000009545 invasion Effects 0.000 abstract 1
- 239000000523 sample Substances 0.000 description 78
- WCUXLLCKKVVCTQ-UHFFFAOYSA-M Potassium chloride Chemical compound [Cl-].[K+] WCUXLLCKKVVCTQ-UHFFFAOYSA-M 0.000 description 74
- 239000000243 solution Substances 0.000 description 59
- 235000011164 potassium chloride Nutrition 0.000 description 37
- 238000012360 testing method Methods 0.000 description 33
- 239000002994 raw material Substances 0.000 description 16
- 239000007788 liquid Substances 0.000 description 14
- 230000000694 effects Effects 0.000 description 10
- 238000011084 recovery Methods 0.000 description 9
- 230000008569 process Effects 0.000 description 7
- 210000004369 blood Anatomy 0.000 description 5
- 239000008280 blood Substances 0.000 description 5
- 210000004027 cell Anatomy 0.000 description 5
- 230000008859 change Effects 0.000 description 5
- 239000002736 nonionic surfactant Substances 0.000 description 4
- 108090000790 Enzymes Proteins 0.000 description 3
- 102000004190 Enzymes Human genes 0.000 description 3
- 229920001503 Glucan Polymers 0.000 description 3
- 239000000470 constituent Substances 0.000 description 3
- 238000004925 denaturation Methods 0.000 description 3
- 230000036425 denaturation Effects 0.000 description 3
- 239000001257 hydrogen Substances 0.000 description 3
- 229910052739 hydrogen Inorganic materials 0.000 description 3
- 239000004615 ingredient Substances 0.000 description 3
- 238000007689 inspection Methods 0.000 description 3
- 239000013010 irrigating solution Substances 0.000 description 3
- 238000005259 measurement Methods 0.000 description 3
- 239000000203 mixture Substances 0.000 description 3
- 239000006228 supernatant Substances 0.000 description 3
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 3
- 241000894006 Bacteria Species 0.000 description 2
- HEDRZPFGACZZDS-UHFFFAOYSA-N Chloroform Chemical compound ClC(Cl)Cl HEDRZPFGACZZDS-UHFFFAOYSA-N 0.000 description 2
- 206010017533 Fungal infection Diseases 0.000 description 2
- 241000239218 Limulus Species 0.000 description 2
- 208000031888 Mycoses Diseases 0.000 description 2
- 230000003213 activating effect Effects 0.000 description 2
- 229910052783 alkali metal Inorganic materials 0.000 description 2
- 150000001340 alkali metals Chemical class 0.000 description 2
- 230000008901 benefit Effects 0.000 description 2
- 210000000601 blood cell Anatomy 0.000 description 2
- 238000004364 calculation method Methods 0.000 description 2
- 238000004090 dissolution Methods 0.000 description 2
- 238000010438 heat treatment Methods 0.000 description 2
- 239000006210 lotion Substances 0.000 description 2
- 239000003580 lung surfactant Substances 0.000 description 2
- 239000006166 lysate Substances 0.000 description 2
- 229920002521 macromolecule Polymers 0.000 description 2
- 230000001376 precipitating effect Effects 0.000 description 2
- 238000004321 preservation Methods 0.000 description 2
- 238000002203 pretreatment Methods 0.000 description 2
- 238000005406 washing Methods 0.000 description 2
- JGSARLDLIJGVTE-UHFFFAOYSA-N 3,3-dimethyl-7-oxo-6-[(2-phenylacetyl)amino]-4-thia-1-azabicyclo[3.2.0]heptane-2-carboxylic acid Chemical compound O=C1N2C(C(O)=O)C(C)(C)SC2C1NC(=O)CC1=CC=CC=C1 JGSARLDLIJGVTE-UHFFFAOYSA-N 0.000 description 1
- 241000228212 Aspergillus Species 0.000 description 1
- OYPRJOBELJOOCE-UHFFFAOYSA-N Calcium Chemical compound [Ca] OYPRJOBELJOOCE-UHFFFAOYSA-N 0.000 description 1
- 241000222122 Candida albicans Species 0.000 description 1
- QCDFBFJGMNKBDO-UHFFFAOYSA-N Clioquinol Chemical compound C1=CN=C2C(O)=C(I)C=C(Cl)C2=C1 QCDFBFJGMNKBDO-UHFFFAOYSA-N 0.000 description 1
- UFHFLCQGNIYNRP-UHFFFAOYSA-N Hydrogen Chemical compound [H][H] UFHFLCQGNIYNRP-UHFFFAOYSA-N 0.000 description 1
- DGAQECJNVWCQMB-PUAWFVPOSA-M Ilexoside XXIX Chemical compound C[C@@H]1CC[C@@]2(CC[C@@]3(C(=CC[C@H]4[C@]3(CC[C@@H]5[C@@]4(CC[C@@H](C5(C)C)OS(=O)(=O)[O-])C)C)[C@@H]2[C@]1(C)O)C)C(=O)O[C@H]6[C@@H]([C@H]([C@@H]([C@H](O6)CO)O)O)O.[Na+] DGAQECJNVWCQMB-PUAWFVPOSA-M 0.000 description 1
- 206010061218 Inflammation Diseases 0.000 description 1
- WHXSMMKQMYFTQS-UHFFFAOYSA-N Lithium Chemical compound [Li] WHXSMMKQMYFTQS-UHFFFAOYSA-N 0.000 description 1
- FYYHWMGAXLPEAU-UHFFFAOYSA-N Magnesium Chemical compound [Mg] FYYHWMGAXLPEAU-UHFFFAOYSA-N 0.000 description 1
- ZLMJMSJWJFRBEC-UHFFFAOYSA-N Potassium Chemical compound [K] ZLMJMSJWJFRBEC-UHFFFAOYSA-N 0.000 description 1
- 229910000272 alkali metal oxide Inorganic materials 0.000 description 1
- 102000015395 alpha 1-Antitrypsin Human genes 0.000 description 1
- 108010050122 alpha 1-Antitrypsin Proteins 0.000 description 1
- 229940024142 alpha 1-antitrypsin Drugs 0.000 description 1
- 239000003708 ampul Substances 0.000 description 1
- 238000004458 analytical method Methods 0.000 description 1
- 239000007864 aqueous solution Substances 0.000 description 1
- QVGXLLKOCUKJST-UHFFFAOYSA-N atomic oxygen Chemical compound [O] QVGXLLKOCUKJST-UHFFFAOYSA-N 0.000 description 1
- 230000004888 barrier function Effects 0.000 description 1
- 238000005842 biochemical reaction Methods 0.000 description 1
- 230000003115 biocidal effect Effects 0.000 description 1
- 210000001124 body fluid Anatomy 0.000 description 1
- 239000010839 body fluid Substances 0.000 description 1
- 229910052791 calcium Inorganic materials 0.000 description 1
- 239000011575 calcium Substances 0.000 description 1
- 229940095731 candida albicans Drugs 0.000 description 1
- 239000013592 cell lysate Substances 0.000 description 1
- 210000002421 cell wall Anatomy 0.000 description 1
- 210000001175 cerebrospinal fluid Anatomy 0.000 description 1
- 238000004140 cleaning Methods 0.000 description 1
- 230000001112 coagulating effect Effects 0.000 description 1
- 238000005345 coagulation Methods 0.000 description 1
- 230000015271 coagulation Effects 0.000 description 1
- 230000000052 comparative effect Effects 0.000 description 1
- 210000000172 cytosol Anatomy 0.000 description 1
- 238000000354 decomposition reaction Methods 0.000 description 1
- 230000007850 degeneration Effects 0.000 description 1
- 238000011161 development Methods 0.000 description 1
- 230000018109 developmental process Effects 0.000 description 1
- 238000003745 diagnosis Methods 0.000 description 1
- 238000003113 dilution method Methods 0.000 description 1
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 238000002474 experimental method Methods 0.000 description 1
- 238000000605 extraction Methods 0.000 description 1
- 239000003102 growth factor Substances 0.000 description 1
- 230000026030 halogenation Effects 0.000 description 1
- 238000005658 halogenation reaction Methods 0.000 description 1
- 230000012447 hatching Effects 0.000 description 1
- 238000000265 homogenisation Methods 0.000 description 1
- 150000002431 hydrogen Chemical class 0.000 description 1
- XLYOFNOQVPJJNP-UHFFFAOYSA-M hydroxide Chemical compound [OH-] XLYOFNOQVPJJNP-UHFFFAOYSA-M 0.000 description 1
- 239000003018 immunosuppressive agent Substances 0.000 description 1
- 239000012535 impurity Substances 0.000 description 1
- 208000033065 inborn errors of immunity Diseases 0.000 description 1
- 208000015181 infectious disease Diseases 0.000 description 1
- 230000004054 inflammatory process Effects 0.000 description 1
- 150000002500 ions Chemical class 0.000 description 1
- 150000002632 lipids Chemical class 0.000 description 1
- 229910052744 lithium Inorganic materials 0.000 description 1
- 230000007774 longterm Effects 0.000 description 1
- 210000004072 lung Anatomy 0.000 description 1
- 229910052749 magnesium Inorganic materials 0.000 description 1
- 239000011777 magnesium Substances 0.000 description 1
- 244000005700 microbiome Species 0.000 description 1
- 210000000056 organ Anatomy 0.000 description 1
- 230000008520 organization Effects 0.000 description 1
- 238000013021 overheating Methods 0.000 description 1
- 239000001301 oxygen Substances 0.000 description 1
- 229910052760 oxygen Inorganic materials 0.000 description 1
- 238000012856 packing Methods 0.000 description 1
- 210000001539 phagocyte Anatomy 0.000 description 1
- 230000008092 positive effect Effects 0.000 description 1
- 229910052700 potassium Inorganic materials 0.000 description 1
- 239000011591 potassium Substances 0.000 description 1
- 238000001556 precipitation Methods 0.000 description 1
- 238000002360 preparation method Methods 0.000 description 1
- 125000002924 primary amino group Chemical group [H]N([H])* 0.000 description 1
- 208000028529 primary immunodeficiency disease Diseases 0.000 description 1
- 239000000047 product Substances 0.000 description 1
- 238000000197 pyrolysis Methods 0.000 description 1
- 230000004044 response Effects 0.000 description 1
- 239000013049 sediment Substances 0.000 description 1
- 238000000926 separation method Methods 0.000 description 1
- 229910052708 sodium Inorganic materials 0.000 description 1
- 239000011734 sodium Substances 0.000 description 1
- 238000001228 spectrum Methods 0.000 description 1
- 239000008223 sterile water Substances 0.000 description 1
- AHBGXTDRMVNFER-UHFFFAOYSA-L strontium dichloride Chemical compound [Cl-].[Cl-].[Sr+2] AHBGXTDRMVNFER-UHFFFAOYSA-L 0.000 description 1
- 238000013268 sustained release Methods 0.000 description 1
- 239000012730 sustained-release form Substances 0.000 description 1
- 230000002195 synergetic effect Effects 0.000 description 1
- 210000001519 tissue Anatomy 0.000 description 1
- 238000012795 verification Methods 0.000 description 1
Classifications
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
- G01N33/569—Immunoassay; Biospecific binding assay; Materials therefor for microorganisms, e.g. protozoa, bacteria, viruses
- G01N33/56961—Plant cells or fungi
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
- G01N33/531—Production of immunochemical test materials
Landscapes
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Immunology (AREA)
- Engineering & Computer Science (AREA)
- Hematology (AREA)
- Chemical & Material Sciences (AREA)
- Urology & Nephrology (AREA)
- Molecular Biology (AREA)
- Biomedical Technology (AREA)
- Cell Biology (AREA)
- Microbiology (AREA)
- Physics & Mathematics (AREA)
- Biochemistry (AREA)
- Pathology (AREA)
- General Physics & Mathematics (AREA)
- Biotechnology (AREA)
- Food Science & Technology (AREA)
- Medicinal Chemistry (AREA)
- General Health & Medical Sciences (AREA)
- Analytical Chemistry (AREA)
- Mycology (AREA)
- Botany (AREA)
- Tropical Medicine & Parasitology (AREA)
- Virology (AREA)
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
Abstract
The invention discloses a kind of bronchoalveolar lavage fluid sample treatment solution and processing detection methods, are made of inorganic salts and surfactant, which can reduce other substances in bronchoalveolar lavage fluid and interfere fungi (1-3)-callose detection.The present invention is handled bronchoalveolar lavage fluid sample, for fungi (1-3)-callose content in vitro detection invasion Pulmonary Fungal Infections patient's bronchoalveolar lavage fluid, compared with the method for traditional serum detection, its detection sensitivity is higher, and is able to maintain higher specificity and accuracy.
Description
Technical field
The invention belongs to fungi (1-3)-callose vitro detection fields, fill more particularly, to a kind of alveolar
Washing lotion sample treatment solution and processing detection method.
Background technique
The present invention is used for auxiliary diagnosis clinically aggressive deep fungal infection.With cortin, immunosuppressor,
Largely using for various wide spectrum class antibiotic and continually developing for the clinical technologies such as organ transplant, for some hypoimmunities
Patient, the probability of invasive infections with fungi can generally increase.(1-3)-β-D glucan is medically most of important fungi
The cell wall constituent of (including candida albicans and Aspergillus), human body phagocyte swallow after fungi can the sustained release substance,
Increase its concentration in the human body fluids such as blood, bronchoalveolar lavage fluid, cerebrospinal fluid, therefore (1-3)-β-D glucan is detected as
One critical index.Currently, G testing inspection sample is mostly serum, and in comparison, aggressive Pulmonary Fungal Infections
Patient, only portion of dextran enter blood, and content is far below the content in bronchoalveolar lavage fluid, so for aggressive Fungal infection of the lung
Infected patient, detects bronchoalveolar lavage fluid, and recall rate is higher.
G test alkali process method due to it detects that (1-3)-β-D glucan high sensitivity, specificity are high, detection time is short by
Hospital uses.Hospital mostly uses heating dilution method to carry out pre-treatment to sample before, but the method is difficult to make in sample (1-3)-β-
D glucan form (single stranded form, bifilar helix or triple helix form) homogenization, to influence the inspection of (1-3)-β-D glucan
Specificity is surveyed, foreign scholar is studies have shown that (1-3)-β-D glucan can be changed into triple helix after Overheating Treatment, and three
Stock helical structure can be hydrolyzed to single-stranded structure through alkali process.Therefore this invention test using G test alkali process method and be ground
Study carefully.
The ingredient of bronchoalveolar lavage fluid includes cell, pulmonary surfactant (pulmonary surfactant, PS), inflammation
Disease medium, growth factor, each hatching egg matter, enzyme, lipid etc..In bronchoalveolar lavage fluid sample component there are certain interference because
Son can seriously affect the measurement result of (1-3)-β-D glucan, reduce the specificity and accuracy of measurement.So-called reaction is dry
Disturbing the factor refers to the response factor unrelated with fixed biochemical reaction (the false positive factor) either in the either phase of reaction
There is the factor (the false negative factor) of interference effect to the reaction.It reacts and contains such factor in sample, that is, bronchoalveolar lavage fluid.Such as
Xa factor shows the effect similar with the coagulating enzyme in lysate, therefore becomes the false positive factor.Alpha1-antitrypsin
There is very strong interference effect to reaction, therefore becomes the false negative factor.
The effect of alkali metal hydroxide is to make to react interference factor denaturation.Highly basic is broken the hydrogen bond of protein,
Also can also there are the fracture and generation of chemical bond in change procedure with the forming salts such as free amino simultaneously.Interference after denaturation because
Son is still existed in solution with the state of dissolution.And heating is diluted only by simple physical change, so that albuminous degeneration is heavy
It forms sediment, the albumen precipitation after denaturation is possible to and can carry out the interference on turbidity to reaction zone.KCl is to glucan to blood constituent
It adsorbs inhibited.The addition of alkali halide, prevents glucan to be adsorbed on blood constituent, so that G-factor grade
Connection reaction is smoothly activated by glucan, so that reaction system stabilizes, has the function of improving reproducibility.But simple use
Treatment fluid used in traditional serum detection is not sufficient to completely remove the interference factor in sample, testing result accuracy
Also to be improved, therefore, the interference problem solved in bronchoalveolar lavage fluid detection is particularly important, it avoids patient's later period need not
The clinical examination and unreasonable treatment wanted.
Summary of the invention
To solve to detect interference problem, the present invention provides a kind of bronchoalveolar lavage fluid sample treatment solution and processing detection sides
Method, using G test alkali process method, eliminate or reduce alveolar sample in influence fungi (1-3)-β-D glucan detection interference because
Element, to achieve the purpose that improve fungi (1-3)-β-D glucan detection sensitivity and specificity.
The technical solution adopted by the present invention is that: a kind of bronchoalveolar lavage fluid sample treatment solution, it is characterised in that: including processing
Liquid A and treatment fluid B, wherein treatment fluid A includes alkali metal hydroxide and surfactant, and treatment fluid B includes alkali metal halogenation
Object.
Preferably, alkali metal hydroxide is one of hydrogen LiOH, NaOH, KOH.
Preferably, alkali halide KCl, MgCl2、CaCl2、SrCl2One of.
Preferably, surfactant is Triton series or one of Tween series, preferably Triton X-100 or
Tween 20。
Preferably, treatment fluid A specifically includes KCl 0.05-0.5M, Triton X-1000.01-1% or Tween 20
0.50-1.50%, treatment fluid B specifically include KOH 0.05-0.5M.
Preferably, treatment fluid A includes KCl 0.5M, and Triton X-100 0.10%, treatment fluid B include KOH 0.25M.
A method of fungi is detected using sample treatment solution pretreatment bronchoalveolar lavage fluid, the specific steps are as follows:
Step 1: configuration treatment fluid A, including KCl 0.05-0.5M, Triton X-100 0.01-1%, configuration processing
Liquid B, is KOH 0.05-0.5M, and treatment fluid A is mixed in equal volume with treatment fluid B Ji Wei sample treatment solution；
Step 2: sample treatment solution is added into bronchoalveolar lavage fluid sample, is incubated for after mixing；
Step 3: the reaction main agent for detecting (1-3)-β-D glucan being added into the mixed liquor after incubation, concussion
It is detected after mixing and calculates (1-3)-β-D beta-dextran content.
Preferably, sample treatment solution specifically includes KCl 0.5M, KOH 0.25M, Triton X-100 0.10%.
Preferably, bronchoalveolar lavage fluid sample and sample treatment solution volume ratio are 1:3-5, preferably 1:4 in step 2.
Preferably, incubation temperature is 37 DEG C in step 2, incubation time 10-15min.
Preferably, the reaction main agent used in step 3 includes G-factor, coagulase source and coagulated protein source, alveolar wass
Liquid and reaction main agent volume ratio are 1:15-25, preferably 1:20.
The advantages and positive effects of the present invention are:
1 this programme is for detecting alveolar cleaning solution, (1-3)-for aggressive Pulmonary Fungal Infections patient, in serum
β-D beta-dextran content will detect bronchoalveolar lavage fluid, recall rate is higher, can judge to invade earlier well below bronchoalveolar lavage fluid
Attacking property Pulmonary Fungal Infections patient；
2 bronchoalveolar lavage fluid samples are contained more interference factors, are related to using this programme compared with serum sample
Sample treatment solution can effectively remove these interference factors, keep testing result more accurate, improve the specificity of measurement
And accuracy；
The surfactant of 3 low concentrations can help decomposition of protein enzyme, reduce the interference factor of bronchoalveolar lavage fluid to detection
Influence, with alkali metal hydroxide and alkali halide cooperation can be improved detection specificity；
4 detection schemes are easy to operate, and detection rates are high, can be quickly obtained testing result.
Specific embodiment
It explains below to the embodiment of the present invention.
The present invention relates to a kind of bronchoalveolar lavage fluid sample treatment solution, sample treatment solution includes at least following components: alkali metal
Hydroxide, alkali halide and surfactant.Alkali metal hydroxide has the hydroxide of the alkali metal such as lithium, sodium, potassium
Object, specifically lithium hydroxide (LiOH), sodium hydroxide (NaOH), potassium hydroxide (KOH).Treatment fluid usually uses one
The aqueous solution of kind or a variety of alkali metal hydroxides, compares through overtesting, uses its detection effect of NaOH or KOH and economic effect
Benefit is higher.Alkali halide has potassium chloride, magnesium chloride, calcium chloride, strontium chloride etc., but due to magnesium, calcium plasma easily with hydrogen
Oxygen root knot, which is closed, generates precipitating, and it is advantageous to potassium chloride as treatment fluid.Nonionic surfactant select Triton series or
Tween series.
Treatment fluid of the invention can be divided into two kinds of solution and save, and when in use again mix these solution.It is false
As alkali metal oxide and alkali metal hydroxide or nonionic surface active agent are mixed and long-term preservation, just
The improper situations such as precipitating can be generated, therefore their separated preservations need to be mixed again when will carry out pre-treatment
It closes.So it includes alkali halide and nonionic surfactant, place that bronchoalveolar lavage fluid sample treatment solution, which is divided into treatment fluid A,
Managing liquid B includes alkali metal hydroxide.
A kind of bronchoalveolar lavage fluid sample treatment solution, including treatment fluid A and treatment fluid B, wherein treatment fluid A includes KCl and table
Face activating agent, wherein treatment fluid includes KCl 0.05-0.5M, surfactant 0.01-1%；Treatment fluid B is KOH solution, tool
Body is KOH 0.05-0.5M.Triton X-100 or Tween20 can be used in surfactant.
It is found by a large amount of comparative tests, treatment fluid A is KCl 0.5M, and Triton X-100 0.10%, treatment fluid B are
When KOH 0.25M, treatment effect is best.
A method of fungi is detected using sample treatment solution pretreatment bronchoalveolar lavage fluid, the specific steps are as follows:
Step 1: configuration treatment fluid A, including KCl 0.05-0.5M, Triton X-100/Tween20 0.01-1%,
Treatment fluid B is configured, is KOH 0.05-0.5M, treatment fluid A is mixed in equal volume with treatment fluid B Ji Wei sample treatment solution；
Step 2: sample treatment solution is added into bronchoalveolar lavage fluid sample, addition volume ratio is 1:3-5,37 after mixing
DEG C be incubated for 10-15min；
Step 3: the reaction main agent for detecting (1-3)-β-D glucan, alveolar are added into the mixed liquor after incubation
Irrigating solution and reaction main agent volume ratio are 1:15-25, and concussion detects after mixing and calculates (1-3)-β-D beta-dextran content.
When bronchoalveolar lavage fluid sample and sample treatment solution volume ratio are 1:4, bronchoalveolar lavage fluid sample and reaction main agent volume
When than for 1:20, test efficiency is higher.
Wherein, the preparation step of reaction main agent is as follows
1 separation and Extraction east limulus blood cell lysate, sterile working take about 20g haemocyte, and blood is added according to 1-5 ratio
Cell pyrolysis liquid, and 200000rpm/min is carried out with high speed pulp grinder, 5-10 minutes damaged cells are thin by the tissue after breakage
Cytosol is put into be rapidly frozen in -30 DEG C of refrigerators and save 10 hours, later takes out frozen cell liquid, is slowly dissolved at room temperature,
200000rpm/min is carried out after all dissolutions, then with the equal pulp grinder of high-speed organization, 5-10 minutes damaged cells then will be damaged
Histocyte liquid afterwards is placed again into be rapidly frozen in -30 DEG C of refrigerators and save 10 hours, repeats last time process 2 times.
2 will carry out 3000rpm/min after the taking-up of above-mentioned haemocyte lysate, are centrifuged within 5-8 minutes, take supernatant, according to 1:
Chloroform is added in 2 ratios, and acutely concussion 10 minutes, are then transferred in apyrogeneity centrifugal precipition tube and carry out at 4 DEG C
It separates within 10000rpm/min, 5-10 minutes, it is spare carefully to take out supernatant.
The supernatant of the 3 east limulus blood cells for taking extracting and developing good is added to molten containing 1.0M magnesium chloride 0.5M calcium chloride
In liquid, packing is freeze-dried into cillin bottle or ampoule bottle as reaction main agent.
The reaction main agent that this programme uses can also directly adopt in commercially available (1-3)-β-D glucan serum detection kit
Reaction main agent, for example, by using fungi (1-3)-β-D glucan detection kit of Tianjin Xi Nuo biological medicine Co., Ltd
Reaction main agent in (development process).
Detection liquid after mixing in step 3 can pass through microorganism quick dynamic detection system (MB-80A, MB-80M, MB-
80S, MB-80X), fungi/dynamic of bacteria detector (MB-80A, MB-80X, MB-80M) or full-automatic fungi/dynamic of bacteria
Detector (IGL-800) is detected, and following embodiment is detected by the comparison of detection data and detectable substance cultivation results
The specificity and accuracy of scheme.
Doing the optional concentration range of sample treatment solution for treatment fluid B, KOH is 0.05-0.5M, in order to further determine KOH
Optimum concentration only includes KOH with treatment fluid, and 100pg/mL fungi standard items are added into alveolar sample, detects various concentration
KOH, the influence to the sample addition mark product test rate of recovery, testing result are as shown in table 1.
Table 1
It can be seen that by test result, only with sterile water process sample, inspection does not measure beta-dextran content, as KOH concentration increases
Add, the rate of recovery gradually increases, and for KOH concentration in 0.05-0.40M, the rate of recovery shows testing result within 50%-150%
It is significant.In conjunction with sample cultivation results, KOH concentration testing result in 0.15-0.50M is feminine gender, therefore KOH concentration is in 0.15-
It is more suitable for handling bronchoalveolar lavage fluid sample when 0.40M.Table 1 statistics indicate that, when treatment fluid only has KOH, the rate of recovery is not
Height illustrates to interfere there are still certain, adds alkali halide and nonionic surfactant that can improve rate of recovery drop in treatment fluid
Low interference.The concentration that KOH is fixed in later detection selects an intermediate concentration 0.25M, carries out to other compositions effect
Analysis.
Embodiment 1
A kind of bronchoalveolar lavage fluid sample treatment solution, including following raw material: treatment fluid A (KCl 0.12M, Triton X-
100 0.01%), treatment fluid B (KOH 0.25M).
Use the method for sample treatment solution pretreatment bronchoalveolar lavage fluid detection fungi, the specific steps are as follows:
Step 1: treatment fluid A is respectively configured and configures treatment fluid B, treatment fluid A is mixed in equal volume with treatment fluid B Ji Wei sample
Present treatment liquid；
Step 2: 40 μ L of sample treatment solution being added into 10 μ L bronchoalveolar lavage fluid samples, is incubated for after mixing at 37 DEG C
10min；
Step 3: 200 μ L reaction main agents being added into the mixed liquor after incubation, and concussion detects after mixing and calculates (1-
3)-β-D beta-dextran content.
Embodiment 2
A kind of bronchoalveolar lavage fluid sample treatment solution, including following raw material: treatment fluid A (KCl 0.12M, Triton X-
100 0.10%), treatment fluid B (KOH 0.25M).
Use the method such as embodiment 1 of sample treatment solution pretreatment bronchoalveolar lavage fluid detection fungi.
Embodiment 3
A kind of bronchoalveolar lavage fluid sample treatment solution, including following raw material: treatment fluid A (KCl 0.12M, Triton X-
100 0.50%), treatment fluid B (KOH 0.25M).
Use the method such as embodiment 1 of sample treatment solution pretreatment bronchoalveolar lavage fluid detection fungi.
Embodiment 4
A kind of bronchoalveolar lavage fluid sample treatment solution, including following raw material: treatment fluid A (KCl 0.12M, Triton X-
100 1.00%), treatment fluid B (KOH 0.25M).
Use the method such as embodiment 1 of sample treatment solution pretreatment bronchoalveolar lavage fluid detection fungi.
Embodiment 5
A kind of bronchoalveolar lavage fluid sample treatment solution, including following raw material: treatment fluid A (KCl 0.06M, Triton X-
100 0.10%), treatment fluid B (KOH 0.25M).
Use the method such as embodiment 1 of sample treatment solution pretreatment bronchoalveolar lavage fluid detection fungi.
Embodiment 6
A kind of bronchoalveolar lavage fluid sample treatment solution, including following raw material: treatment fluid A (KCl 0.12M, Triton X-
100 0.10%), treatment fluid B (KOH 0.25M).
Use the method such as embodiment 1 of sample treatment solution pretreatment bronchoalveolar lavage fluid detection fungi.
Embodiment 7
A kind of bronchoalveolar lavage fluid sample treatment solution, including following raw material: treatment fluid A (KCl 0.50M, Triton X-
100 0.10%), treatment fluid B (KOH 0.25M).
Use the method such as embodiment 1 of sample treatment solution pretreatment bronchoalveolar lavage fluid detection fungi.
Embodiment 8
A kind of bronchoalveolar lavage fluid sample treatment solution, including following raw material: treatment fluid A (KCl 0.50M, Triton X-
100 0.10%), treatment fluid B (KOH 0.25M).
Use the method such as embodiment 1 of sample treatment solution pretreatment bronchoalveolar lavage fluid detection fungi.
Embodiment 9
A kind of bronchoalveolar lavage fluid sample treatment solution, including following raw material: treatment fluid A (KCl 0.50M), treatment fluid B
(KOH 0.25M)。
Use the method such as embodiment 1 of sample treatment solution pretreatment bronchoalveolar lavage fluid detection fungi.
Embodiment 10
A kind of bronchoalveolar lavage fluid sample treatment solution, including following raw material: treatment fluid A (Triton X-100 0.10%),
Treatment fluid B (KOH 0.25M).
Use the method such as embodiment 1 of sample treatment solution pretreatment bronchoalveolar lavage fluid detection fungi.
Embodiment 11
A kind of bronchoalveolar lavage fluid sample treatment solution, including following raw material: treatment fluid A (KCl 0.50M, Triton X-
100 0.05%), treatment fluid B (KOH 0.25M).
Use the method such as embodiment 1 of sample treatment solution pretreatment bronchoalveolar lavage fluid detection fungi.
Embodiment 12Tween20 detects bronchoalveolar lavage fluid as treatment fluid as surfactant
A kind of bronchoalveolar lavage fluid sample treatment solution, including following raw material: treatment fluid A (KCl 0.50M, Tween20
0.10%), treatment fluid B (KOH 0.25M).
Use the method such as embodiment 1 of sample treatment solution pretreatment bronchoalveolar lavage fluid detection fungi.
Embodiment 13
A kind of bronchoalveolar lavage fluid sample treatment solution, including following raw material: treatment fluid A (KCl 0.50M, Tween20
0.50%), treatment fluid B (KOH 0.25M).
Use the method such as embodiment 1 of sample treatment solution pretreatment bronchoalveolar lavage fluid detection fungi.
Embodiment 14
A kind of bronchoalveolar lavage fluid sample treatment solution, including following raw material: treatment fluid A (KCl 0.50M, Tween20
1.00%), treatment fluid B (KOH 0.25M).
Use the method such as embodiment 1 of sample treatment solution pretreatment bronchoalveolar lavage fluid detection fungi.
Embodiment 15
A kind of bronchoalveolar lavage fluid sample treatment solution, including following raw material: treatment fluid A (KCl 0.50M, Tween20
1.50%), treatment fluid B (KOH 0.25M).
Use the method such as embodiment 1 of sample treatment solution pretreatment bronchoalveolar lavage fluid detection fungi.
Embodiment 16
A kind of bronchoalveolar lavage fluid sample treatment solution, including following raw material: treatment fluid A (KCl 0.50M, Tween20
2.00%), treatment fluid B (KOH 0.25M).
Use the method such as embodiment 1 of sample treatment solution pretreatment bronchoalveolar lavage fluid detection fungi.
Embodiment 1-4 is wherein compared, the results are shown in Table 2.
Table 2
In embodiment 1-4, treatment fluid B component is constant, and KCl concentration is constant in treatment fluid A, wherein Triton X-100 concentration
It gradually rises, for Triton X-100 in 0.1%-1.0% change procedure, pattern detection concentration is first to reduce to increase afterwards, can be bright
It is aobvious to find out that for Triton X-100 concentration at 0.1%, testing result is closest with cultivation results in embodiment 2, remove alveolar
The dynamics of interference factor is maximum in irrigating solution, and effect is best.
Embodiment 5-7 is compared, the results are shown in Table 3.
Table 3
In embodiment 5-7, treatment fluid B component is constant, and Triton X-100 concentration is constant in treatment fluid A, KCl concentration by
Edge up height, it can be seen that prescription for the treatment of liquid detects several sample results and is consistent with cultivation results in embodiment 7, removes alveolar
The dynamics of interference factor is maximum in irrigating solution, and effect is best, so when KCl concentration is selected as 0.50M in treatment fluid A, detection spirit
Sensitivity is best.
Embodiment 8-10 is compared, the results are shown in Table 4.
Table 4
In embodiment 8-10, treatment fluid B component is constant, be respectively set as in treatment fluid A comprising Triton X-100 and
KCl, only KCl, only Triton X-100, compare its accuracy in detection, can be found out embodiment 8 according to test result
Middle KCl and Triton X-100 are existed simultaneously when detecting in treatment fluid A, and testing result more meets with cultivation results, and
The only testing result of the treatment fluid A including KCl or Triton X-100, error rate is higher, thus proves, fills for alveolar
For washing lotion as detection sample, KCl and Triton X-100 are indispensable.The solution that KCl is only added is low molecule decentralized system,
Main dispersed phase is molecule, ion, but for other macromolecular substances in bronchoalveolar lavage fluid, does not have peptizaiton；Surface
Activating agent be dissolved in water energy enough and significantly reduce water surface can, when low concentration, also can significantly reduce surface tension, be able to solve sample
The problem of middle macromolecular substances are reunited, and verified and found by a large number of experiments, the addition of nonionic surfactant is formed
Steric hindrance reduces the generation of reunion, good space barrier is not easy dextran molecule so that three-dimensional system is stablized
In other big molecular impurities occur coagulation phenomena such as, therefore only both synergistic effect just can guarantee testing result with compared with
High-accuracy.
Embodiment 8 and embodiment 11 are compared, the results are shown in Table 5.
Table 5
Embodiment 8 and embodiment 11 have used the Triton X- of 0.10% Triton X-100 and 0.05% respectively
100, it is compared by the test result, the obvious accuracy rate of embodiment 8 is higher, occurs some false positive results in embodiment 11, real
Apply 8 treatment fluid A ingredient of example be 0.5M KCl, 0.05% Triton X-100 when, this formula treatment fluid reduces interference, survey
Test result is more matched with cultivation results.
Embodiment 8 and embodiment 12-16 are compared, the results are shown in Table 6.
Table 6
Upper table statistics indicate that, in 0.50-1.50%, testing result is consistent Tween20 concentration with example 8 (control), and
Matched with cultivation results, illustrate Tween20 can effectively be removed as one for the treatment of fluid ingredient interfered in bronchoalveolar lavage fluid because
Son improves the sensitivity and specificity of bronchoalveolar lavage fluid testing result.
The detection of 17 sensitivity and specificity of embodiment
Prescription for the treatment of liquid in selection example 8 is after control test is handled using treatment fluid using happiness promise with cultivation results
The sensitivity and specificity (selection 100 has the BALF sample of culture information to be tested) of G test kit detection,
Like the judgement of promise G test kit testing result:
| It is negative | ≤60pg/mL |
| Gray area | 60-100pg/mL |
| It is positive | ≥100pg/mL |
Sensitivity and specificity calculation method such as table 7:
Table 7
Sensitivity: (A+C)/(A+C+E) * 100%
Specificity: (D+F)/(B+D+F) * 100%
Test result such as table 8.
Table 8
Interpretation of result such as table 9
Table 9
It can be seen that by data, treatment fluid composition test alveolar sample results and cultivation results matching degree are high in embodiment 8,
The sensitivity and specificity of the data measured using happiness promise G test kit are up to 90.0% and 92.5%, hence it is evident that improve
Detection accuracy of the kit to bronchoalveolar lavage fluid sample.
Embodiment 18 passes through Detection accuracy after fungi (1-3)-β-D dextran standard rate of recovery verification processing
Prescription for the treatment of liquid in selection example 8, test fungi (1-3)-β-D dextran standard are added to alveolar wass
The rate of recovery in liquid sample, judges Clinical significance of detecting:
Rate of recovery calculation method:
Table 10
As shown in table 10, the measuring rate of recovery of 5 samples illustrates this as a result between 50%-150%
The testing result interference that treatment fluid processing bronchoalveolar lavage fluid obtains is small, and data are credible.
Several embodiments of the present invention are described in detail above, but the content is only preferable reality of the invention
Example is applied, should not be considered as limiting the scope of the invention.It is all according to equivalent change made by the present patent application range with change
Into etc., it should still be within the scope of the patent of the present invention.
Claims (10)
1. a kind of bronchoalveolar lavage fluid sample treatment solution, it is characterised in that: including treatment fluid A and treatment fluid B, wherein treatment fluid A packet
Alkali metal hydroxide and surfactant are included, treatment fluid B includes alkali halide.
2. bronchoalveolar lavage fluid sample treatment solution according to claim 1, it is characterised in that: the alkali metal hydroxide is
One of LiOH, NaOH, KOH.
3. bronchoalveolar lavage fluid sample treatment solution according to claim 1, it is characterised in that: the alkali halide is
KCl、MgCl2、CaCl2、SrCl2One of.
4. bronchoalveolar lavage fluid sample treatment solution according to claim 1, it is characterised in that: the surfactant is
One of Triton series or Tween series, preferably Triton X-100 or Tween 20.
5. bronchoalveolar lavage fluid sample treatment solution according to claim 1, it is characterised in that: treatment fluid A specifically includes KCl
0.05-0.5M, Triton X-1000.01-1% or Tween 20 0.50-1.50%, treatment fluid B specifically include KOH 0.05-
0.5M。
6. bronchoalveolar lavage fluid sample treatment solution according to claim 5, it is characterised in that: treatment fluid A includes KCl 0.5M,
Triton X-100 0.10%, treatment fluid B include KOH 0.25M.
7. a kind of method using sample treatment solution pretreatment bronchoalveolar lavage fluid detection fungi, it is characterised in that: specific steps are such as
Under:
Step 1: configuration treatment fluid A, including KCl 0.05-0.5M, Triton X-100 0.01-1% configure treatment fluid B, are
KOH 0.05-0.5M, treatment fluid A are mixed in equal volume with treatment fluid B Ji Wei sample treatment solution；
Step 2: sample treatment solution is added into bronchoalveolar lavage fluid sample, is incubated for after mixing；
Step 3: the reaction main agent for detecting (1-3)-β-D glucan is added into the mixed liquor after incubation, after concussion mixes
It detects and calculates (1-3)-β-D beta-dextran content；
Preferably sample treatment solution specifically includes KCl 0.5M, KOH 0.25M, Triton X-100 0.10%.
8. the method according to claim 7 using sample treatment solution pretreatment bronchoalveolar lavage fluid detection fungi, feature
Be: bronchoalveolar lavage fluid sample and sample treatment solution volume ratio are 1:3-5, preferably 1:4 in the step 2.
9. the method according to claim 7 or 8 using sample treatment solution pretreatment bronchoalveolar lavage fluid detection fungi, special
Sign is: incubation temperature is 37 DEG C in the step 2, incubation time 10-15min.
10. the method according to claim 7 using sample treatment solution pretreatment bronchoalveolar lavage fluid detection fungi, feature
Be: the reaction main agent used in the step 3 includes G-factor, coagulase source and coagulated protein source, the bronchoalveolar lavage fluid
It is 1:15-25, preferably 1:20 with the reaction main agent volume ratio.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201810622529.5A CN109001455B (en) | 2018-06-15 | 2018-06-15 | Alveolar lavage fluid sample treatment fluid and treatment detection method |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201810622529.5A CN109001455B (en) | 2018-06-15 | 2018-06-15 | Alveolar lavage fluid sample treatment fluid and treatment detection method |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN109001455A true CN109001455A (en) | 2018-12-14 |
| CN109001455B CN109001455B (en) | 2021-09-10 |
Family
ID=64601099
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201810622529.5A Active CN109001455B (en) | 2018-06-15 | 2018-06-15 | Alveolar lavage fluid sample treatment fluid and treatment detection method |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN109001455B (en) |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110823785A (en) * | 2019-11-12 | 2020-02-21 | 北京大学 | Fungus detection method |
Citations (9)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1997446A (en) * | 2004-05-20 | 2007-07-11 | 伊顿研究有限公司 | Compositions containing a hollow glucan particle or a cell wall particle encapsulating a terpene component, methods of making and using them |
| CN102305787A (en) * | 2010-09-08 | 2012-01-04 | 天津市一瑞生物工程有限公司 | Preparation of fungal (1-3)-beta-D-Glucan colorimetric assay kit and using method for colorimetric assay kit |
| US20120244139A1 (en) * | 2005-10-21 | 2012-09-27 | Madison Edwin L | Modified proteases that inhibit complement activation |
| CN103380377A (en) * | 2011-02-25 | 2013-10-30 | 三菱化学美迪恩斯株式会社 | Method for measuring myocardial troponin |
| US20150031568A1 (en) * | 2011-10-26 | 2015-01-29 | Kowa Company, Ltd. | Method for measuring physiologically active substance of biological origin, and microparticles and extract for use in the method |
| 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 |
| US20160061838A1 (en) * | 2012-12-28 | 2016-03-03 | Seikagaku Corporation | Pre-treatment agent and pre-treatment method for antithrombin iii to be subjected to limulus test |
| WO2016086210A1 (en) * | 2014-11-25 | 2016-06-02 | Epiva Biosciences, Inc. | Probiotic and prebiotic compostions, and methods of use therof for treatment and prevention of graft versus host disease |
| CN107466319A (en) * | 2017-06-28 | 2017-12-12 | 丹娜（天津）生物科技有限公司 | GM hybridomas, monoclonal antibody, kit and preparation method and application |
-
2018
- 2018-06-15 CN CN201810622529.5A patent/CN109001455B/en active Active
Patent Citations (9)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1997446A (en) * | 2004-05-20 | 2007-07-11 | 伊顿研究有限公司 | Compositions containing a hollow glucan particle or a cell wall particle encapsulating a terpene component, methods of making and using them |
| US20120244139A1 (en) * | 2005-10-21 | 2012-09-27 | Madison Edwin L | Modified proteases that inhibit complement activation |
| CN102305787A (en) * | 2010-09-08 | 2012-01-04 | 天津市一瑞生物工程有限公司 | Preparation of fungal (1-3)-beta-D-Glucan colorimetric assay kit and using method for colorimetric assay kit |
| CN103380377A (en) * | 2011-02-25 | 2013-10-30 | 三菱化学美迪恩斯株式会社 | Method for measuring myocardial troponin |
| US20150031568A1 (en) * | 2011-10-26 | 2015-01-29 | Kowa Company, Ltd. | Method for measuring physiologically active substance of biological origin, and microparticles and extract for use in the method |
| US20160061838A1 (en) * | 2012-12-28 | 2016-03-03 | Seikagaku Corporation | Pre-treatment agent and pre-treatment method for antithrombin iii to be 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 |
| WO2016086210A1 (en) * | 2014-11-25 | 2016-06-02 | Epiva Biosciences, Inc. | Probiotic and prebiotic compostions, and methods of use therof for treatment and prevention of graft versus host disease |
| CN107466319A (en) * | 2017-06-28 | 2017-12-12 | 丹娜（天津）生物科技有限公司 | GM hybridomas, monoclonal antibody, kit and preparation method and application |
Non-Patent Citations (2)
| Title |
|---|
| MOHAMMAD ASHRAF HOSSAIN 等: ""Comparison between Wako-WB003 and Fungitec G tests for detection of (1→3)-β-D-glucan in systemic mycosis"", 《JOURNAL OF CLINICAL LABORATORY ANALYSIS》 * |
| 张永坚 等: ""β-葡聚糖与内毒素共同作用对细菌内毒素检查的影响"", 《中国药品标准》 * |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110823785A (en) * | 2019-11-12 | 2020-02-21 | 北京大学 | Fungus detection method |
Also Published As
| Publication number | Publication date |
|---|---|
| CN109001455B (en) | 2021-09-10 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| ES2700774T3 (en) | Method for the bacteriological investigation of a biological sample and related device | |
| CN103890584B (en) | For analyzing the individuation quality controls thing of test | |
| CN106198470A (en) | A kind of fungal detection fluorescence staining liquid and application | |
| CN104630325B (en) | A kind of liquid-type fibrinogen detection reagent and preparation method thereof | |
| CN104232739A (en) | Method for detecting bacterial endotoxin in citric acid raw material | |
| CN110734898B (en) | Virus biomagnetic bead enrichment and concentration kit and application | |
| CN109001455A (en) | A kind of bronchoalveolar lavage fluid sample treatment solution and processing detection method | |
| CN114544983A (en) | Preparation method of normal value quality control product for blood coagulation and platelet function analyzer | |
| CN107290545A (en) | β2-microglobulin solution, its preparation method and the application of a kind of stabilization | |
| CN102305787A (en) | Preparation of fungal (1-3)-beta-D-Glucan colorimetric assay kit and using method for colorimetric assay kit | |
| CN112063684A (en) | Method and kit for detecting illegal addition of antibiotics in cosmetics | |
| CN104166004B (en) | Platelet cofactor Ⅰ quality-control product preparation method | |
| CN107271246A (en) | A kind of immune p16 for Thinprep pap testINK4aAntigen preserves liquid and preparation method thereof | |
| CN108344875B (en) | Method for improving sensitivity of reagent for activating partial thromboplastin time to heparin and application | |
| CN109239374A (en) | A kind of liquid thrombin time detection reagent | |
| CN109323996B (en) | Fungus detection kit | |
| CN104677809B (en) | Cryptococcus viability detection kit and detection method | |
| Snyder et al. | Reagents, stains, and media: mycology | |
| US5372935A (en) | Detection of candida | |
| CN112462076B (en) | Method for detecting endotoxin content applied to microcarrier | |
| CN110470849A (en) | A kind of exceptional value quality-control product and preparation method thereof for hemostasis examination | |
| CN102645358A (en) | Preparation method of water-soluble cholesterol lyophilized product derived from serum | |
| Novi et al. | Optimization sonication time and dilution factor in determining the concentration of endotoxin challenge vial with kinetic turbidimetric method | |
| RU2438129C1 (en) | Method of treating hyperparathyreosis | |
| CN105738299B (en) | A kind of method and kit of quick measure intracellular protein relative amount |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| CB02 | Change of applicant information | ||
| CB02 | Change of applicant information |
Address after: 300457 Tianda Science Park C8 3rd Floor, No. 80 Fourth Avenue, Binhai New Area Development Zone, Tianjin Applicant after: TIANJIN ERA BIOLOGY TECHNOLOGY CO.,LTD. Applicant after: TIANJIN XINUO BIOLOGICAL PHARMACEUTICAL Co.,Ltd. Applicant after: BEIJING GOLD MOUNTAINRIVER TECH DEVELOPMENT CO.,LTD. Applicant after: BEIHAI XINGLONG BIOLOGICAL PRODUCT Co.,Ltd. Address before: 300457 Tianda Science Park C8 3rd Floor, No. 80 Fourth Avenue, Binhai New Area Development Zone, Tianjin Applicant before: TIANJIN ERA BIOLOGY TECHNOLOGY CO.,LTD. Applicant before: TIANJIN XINUO BIOLOGICAL PHARMACEUTICAL Co.,Ltd. Applicant before: BEIJING GOLD MOUNTAINRIVER TECH DEVELOPMENT CO.,LTD. Applicant before: BEIHAI XINGLONG BIO-PHARMACY Co.,Ltd. |
|
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant |