CN103792351A - Method for rapidly detecting gram bacteria as well as kit - Google Patents
Method for rapidly detecting gram bacteria as well as kit Download PDFInfo
- Publication number
- CN103792351A CN103792351A CN201410033894.4A CN201410033894A CN103792351A CN 103792351 A CN103792351 A CN 103792351A CN 201410033894 A CN201410033894 A CN 201410033894A CN 103792351 A CN103792351 A CN 103792351A
- Authority
- CN
- China
- Prior art keywords
- gram
- bacteria
- antigen
- antibody
- kit
- 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.)
- Pending
Links
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
- G01N33/56911—Bacteria
-
- 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
- G01N33/58—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving labelled substances
Landscapes
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Engineering & Computer Science (AREA)
- Immunology (AREA)
- Molecular Biology (AREA)
- Biomedical Technology (AREA)
- Chemical & Material Sciences (AREA)
- Hematology (AREA)
- Urology & Nephrology (AREA)
- Cell Biology (AREA)
- Biochemistry (AREA)
- Biotechnology (AREA)
- Food Science & Technology (AREA)
- Medicinal Chemistry (AREA)
- Physics & Mathematics (AREA)
- Analytical Chemistry (AREA)
- Microbiology (AREA)
- General Health & Medical Sciences (AREA)
- General Physics & Mathematics (AREA)
- Pathology (AREA)
- Tropical Medicine & Parasitology (AREA)
- Virology (AREA)
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
Abstract
The invention provides a method for rapidly detecting gram bacteria as well as a kit. The method comprises the following steps: culturing gram bacteria; carrying out low-temperature inactivation treatment on bacteria by nanosecond pulse, and extracting a gram-positive bacteria teichoic acid antigen and a gram-negative bacteria lipopolysaccharide antigen; preparing a colloidal gold solution labeled with goat anti-human IgG; and detecting the existence of the antibody by a dot immunogold filtration technique, thereby judging whether the sample contains a teichoic acid antibody or a lipopolysaccharide antibody. The kit provided by the invention changes disadvantages in traditional microorganism detection, shortens the time for detection process to be within 10 minutes, and also greatly improves the detection flexibility and specificity so as to meet the need of rapid gram bacteria detection.
Description
Technical field
The present invention relates to fast detecting gram-bacteria, method and the kit of the fast detecting gram-bacteria especially obtaining, have significant result of use, and method is very precisely with convenient.
Background technology
Immunological technique is by the specific binding reaction of antigen and antibody, be aided with again immune amplifying technique and carry out discriminating bacteria, the advantage of immunization method is that sample is carrying out after selective enrichment, do not need to separate, can adopt immunological technique to screen, because immunization has higher sensitivity, sample can reach in the short period of time degree of detecting after increasing bacterium, the association reaction of antigen and antibody can complete within very short time, this technology requires also not high to operator, it is the longest Fast Detection Technique the most widely of grass-roots unit's Applicative time so far, wherein immune colloidal gold technique is extremely sensitive one of diagnostic method fast again.
Gram's stain can be divided into two large classes bacterium: all bacteriums that is dyed to purple call gram-positive bacteria; Dye the red Gram-negative bacteria that is called.In experiment, often there will be false positive and false-negative result, false positive is mainly because decolouring is incomplete, may be because smear is blocked up, or violet staining is excessive, cause decolouring not exclusively, false negative may be because cell is excessively fixing, causes the change of cell membrane permeability, and occurs false negative result; In addition, the microbe growth time is oversize, has had the generation death of part bacterium or self-dissolving, also causes the change of cell membrane permeability and occurs false negative result.
Impulse electric field (pulsed electric field, PEF) is the new technology across fields such as bioelectromagnetics, high voltage and microelectronics growing up over the past two decades, is widely used.In field intensity, during up to kV/cm, width can make the lipid bilayer of cell membrane temporarily rearrange in microsecond to the pulse of Millisecond, forms micropore, helps manyly can not pass through smoothly by the large hydrophilic molecular of cell membrane at ordinary times; When cancelling after impulse electric field, micropore can be closed and can not had any impact to cell, and this phenomenon is called electroporation, this technology enters nuclear properties by the film of wearing of ultrashort pulse, the change of having avoided pyrocaloric effect to cause, has effectively kept the activity of membranous antigen, therefore reaches the effect of low temperature deactivation.
The existing many reports of the method for related detection gram-bacteria at present, its process mainly contains portable gram suit and dyes sheet (number of patent application: 201110226527.2, be made up of the Gram stain that concentrating, high osmosis filter paper, high-hydroscopicity sponge and tetrad row polyethylene plastic bag); The deactivation of gram-positive bacteria (number of patent application: 200680035823.1, described method comprises and is exposed to visible ray, and the light within the scope of wavelength 400-500nm especially); Can extract the nucleic acid extraction chip (number of patent application: 201310296807.X, based on the nucleic acid extraction chip of silica bead method extraction gram-positive bacteria and Gram-negative bacteria nucleic acid) of gram-positive bacteria and Gram-negative bacteria nucleic acid; Differentiate nutrient culture media and the using method (number of patent application: 201310412457.9 of Gram-negative and positive bacteria, utilize the character of the specific expressed ALANINE aminopeptidase of Gram-negative bacteria, adopt the substrate of L-Alanine-AMC-TFA as this enzyme, utilize the diffusion of the 4-methylcoumarin of cyclodextrin retardance generation, what produce by bacterium colony on flat board distinguishes Gram-negative bacteria and positive bacteria compared with hyperfluorescenceZeng Yongminggaoyingguang); Gram-positive bacterium method for extracting nucleic acid (number of patent application: 201210162646.0, a kind of reagent of the cell membrane of abolishing gram-positive bacterium is provided, formed by extract A and extract B; By Tris, EDTA, SDS, Tween80, TritonX-100, Brij58,3-[(3-cholesterol aminopropyl) dimethylamino]-1-propane sulfonic acid, sarcosyl, beta-mercaptoethanol and water mixes, and makes extract A; Described extract B is prepared as follows: water-saturated phenol, ethanol and pyrrolidone are mixed, make extract B).
The method of fast detecting gram-bacteria provided by the invention, overcome the defect that false positive and false-negative testing result often appear in existing Gram's stain, testing process was tapered in 10 minutes, and the sensitivity and the specificity that detect are significantly improved, to meet the needs of fast detecting gram-bacteria; Kit provided by the invention, workable simultaneously, has wide market sale prospect.
Summary of the invention
The object of the invention is to: method and the kit of the fast detecting gram-bacteria providing, overcome the defect that false positive and false-negative testing result often appear in existing Gram's stain.
The object of the present invention is achieved like this: a kind of method of fast detecting gram-bacteria, implement step by step;
The first step, gram-bacteria is cultivated: staphylococcus aureus reference culture ATCC25923 and colon bacillus reference culture ATCC25922 are inoculated in respectively on Colombia's blood culture flat board, are placed in 37 ℃, incubator and cultivate after 12h, collect bacterium colony;
Second step, carries out low temperature inactivation treatment with nanosecond pulse to bacterium, extracts gram-positive bacteria LTA antigen and Gram-negative bacteria boivin antigen;
The 3rd step, extracts two kinds of antigens and quality controlled serum point on the nitrocellulose membrane reaction plate preparing by second step;
The 4th step, prepare mark goat anti-human igg's colloidal gold solution: adopt trisodium citrate reduction method to prepare collaurum, the gold chloride that is 0.01% by concentration (purchased from Tianjin chemical reagent factory) solution, putting heating in micro-wave oven boils, add 0.23ml1% sodium citrate (purchased from Tianjin chemical reagent factory) by every 100ml0.01% gold chloride, continue to boil 10～20min, until being shiny red, liquid stops heating, be cooled to after room temperature and return to original volume with ultrapure water, making diameter is the colloid gold particle of 0.22um left and right, by 0.1M sal tartari (purchased from Tianjin chemical reagent factory) solution adjusting collaurum pH value to 6.7, add goat anti-human igg antibody (purchased from Sigma reagent company of the U.S.) to mix by 2.5mg antibody/ml collaurum, the colloid gold particle mark that is 0.22um with above-mentioned diameter by goat anti-human igg antibody obtains.
The 5th step, adopt the existence of dot immunogold diafiltration technology for detection antibody, thereby whether judgement sample contains LTA antibody and lipopolysaccharides antibody.
Second step specifically comprises:
1) prepare gram-positive bacteria membranous antigen: by staphylococcus aureus reference culture ATCC25923, after 37 ℃ of cultivation 12h, collect bacterium colony, process deactivation through the pulse electrical signal of 300ns, 7 groups of concentric cable of consists, high-voltage power supply and electrode composition, build-out resistor is 7 Ω, electric field 20-30kv, ultrasonic degradation after deactivation, under 15000rpm, centrifugal 15min, then through cold phenol extracting, obtain with CNB2 chromatographic column chromatography, after preparing, be stored in-80 ℃ of low temperature and preserve, for subsequent use;
2) prepare Gram-negative bacteria membranous antigen: by colon bacillus reference culture ATCC25922, after 37 ℃ of cultivation 12h, collect bacterium colony, pulse electrical signal through 300ns is processed deactivation, 7 groups of concentric cable of consists, high-voltage power supply and electrode composition, build-out resistor is 7 Ω, electric field 20-30kv, ultrasonic degradation after deactivation, under 15000rpm, centrifugal 15min, again through triton 114 surfactant extractings, heat to 72 ℃, under 15000rpm, centrifugal 15min, filter, get supernatant and cross the acquisition of sephadexG-200 molecular sieve purification, after preparing, being stored in-80 ℃ of low temperature preserves, for subsequent use,
Described gram-positive bacteria membranous antigen is staphylococcus aureus reference culture ATCC25923 LTA antigen, molecular weight 10-100kda; Described Gram-negative bacteria membranous antigen is colon bacillus reference culture ATCC25922 boivin antigen, molecular weight 10-100kda.
The 3rd step specifically comprises:
1) take out after nitrocellulose membrane is soaked to 10min in 10% methyl alcohol, be cut into 1cmx1cm square, film is arranged in the reacting hole of PVC reaction plate that is lined with adsorptive pads with tweezers after clean, 37 ℃ of baking 2h, take out;
On nitrocellulose membrane fixed position, click and enter respectively the Quality Control point that 2 check points of gram-positive bacteria membranous antigen, Gram-negative bacteria membranous antigen and 1 click and enter 30 normal pooled serums with pipettor, point film amount 1-10ul, gram-positive bacteria membranous antigen concentration 0.7-2.2mg/ml, Gram-negative bacteria membranous antigen concentration 0.3-0.8mg/ml; After some film, 37 ℃, baking 1h, put 4 ℃ of stored refrigerated, for subsequent use;
2) 2 check points and 1 the Quality Control point on described nitrocellulose membrane from left to right arranged and formed according to the order of gram-positive bacteria membranous antigen check point, normal serum Quality Control point, Gram-negative bacteria membranous antigen check point successively.
The kit of a kind of fast detecting gram-bacteria provided by the invention, comprises reaction plate, sample dilution, cleansing solution and golden labeling antibody liquid;
Reaction plate is that reaction film is arranged in the reacting hole of the PVC base plate that is lined with adsorptive pads and is made;
Sample dilution: 1mmol/L, Tris-HCL damping fluid 50ml, the polysorbas20 25ul of pH8.5, gelatin 1ml, ultrapure water is settled to 100ml.
Cleansing solution: 1mmol/L, the Tris-HCL damping fluid 50ml of pH8.5, polysorbas20 500ul, Nacl2g, ultrapure water is settled to 100ml.
The mark goat anti-human igg antibody's of preparation colloidal gold solution, goat anti-human igg's concentration 2.5mg/ml, colloid gold particle diameter 0.22um; Colloidal gold solution pH6.7.
The technique of the inventive method be gram-bacteria cultivate → bacterium is carried out to low temperature inactivation treatment with nanosecond pulse, extraction gram-positive bacteria LTA antigen and Gram-negative bacteria boivin antigen → prepare the existence of colloidal gold solution → employing dot immunogold diafiltration technology for detection antibody of mark goat anti-human igg, thus whether judgement sample contains LTA antibody and lipopolysaccharides antibody.Kit of the present invention, has changed the deficiency of traditional Micro biological Tests, shows technical progress.
Embodiment
The invention will be further elaborated by the following examples:
Embodiment
1, prepare gram-positive bacteria LTA antigen
By staphylococcus aureus reference culture ATCC25923(purchased from Huankai Microbes Tech Co., Ltd., Guangdong), cultivate after 12h for 37 ℃ and collect bacterium colony, process deactivation through the pulse electrical signal of 300ns; 7 groups of concentric cable of consists, high-voltage power supply and electrode composition, build-out resistor is 7 Ω, electric field 25kv, ultrasonic degradation after deactivation, under 15000rpm, centrifugal 15min, then obtain with CNB2 chromatographic column (purchased from Sigma reagent company of the U.S.) chromatography through cold phenol extracting, after preparing, being stored in-80 ℃ of low temperature saves backup.
2, prepare Gram-negative bacteria boivin antigen
By colon bacillus reference culture ATCC25922(purchased from Huankai Microbes Tech Co., Ltd., Guangdong), after 37 ℃ of cultivation 12h, collect bacterium colony, pulse electrical signal through 300ns is processed deactivation, 7 groups of concentric cable of consists, high-voltage power supply and electrode composition, build-out resistor is 7 Ω, electric field 25kv, ultrasonic degradation after deactivation, under 15000rpm, centrifugal 15min, again through triton 114 surfactants (purchased from Sigma reagent company of the U.S.) extracting, heat to 72 ℃, again under 15000rpm, centrifugal 15min, filter, get supernatant and cross the acquisition of sephadexG-200 molecular sieve (purchased from Sigma reagent company of the U.S.) purifying, after preparing, being stored in-80 ℃ of low temperature saves backup.
3, the preparation of reaction plate
After being soaked to 10min in 10% methyl alcohol, takes out nitrocellulose membrane (purchased from Whatman company of Britain), be cut into the square of 1cmx1cm, with tweezers after clean, film is arranged on to the PVC reaction plate that is lined with adsorptive pads (purchased from the outstanding Bioisystech Co., Ltd in Shanghai) and (selects PVC starting material, make the reaction plate of 5cm*3cm through injection molding, in the middle of this reaction plate, have 0.8cm*0.8cm reacting hole) in, 37 ℃ of baking 2h, take out for subsequent use;
On nitrocellulose membrane fixed position, click and enter respectively gram-positive bacteria LTA antigen, 30 pooled serums, Gram-negative bacteria boivin antigen according to order from left to right with pipettor; Point film amount 10ul, gram-positive bacteria LTA antigen concentration 1.4mg/ml, Gram-negative bacteria boivin antigen concentration 0.5mg/ml; After some film, 37 ℃ of baking 1h, put 4 ℃ of stored refrigerated for subsequent use.
4, sample dilution: 1mmol/L, Tris-HCL damping fluid 50ml, the polysorbas20 25ul of pH8.5, gelatin 1ml, ultrapure water is settled to 100ml.
5, cleansing solution: cleansing solution: 1mmol/L, the Tris-HCL damping fluid 50ml of pH8.5, polysorbas20 500ul, Nacl2g, ultrapure water is settled to 100ml.
6, the preparation of golden labeling antibody liquid
Adopt trisodium citrate reduction method to prepare collaurum, the gold chloride that is 0.01% by concentration (purchased from Tianjin chemical reagent factory) solution, putting heating in micro-wave oven boils, add 0.23ml1% sodium citrate (purchased from Tianjin chemical reagent factory) by every 100ml0.01% gold chloride, continue to boil 10～20min, until being shiny red, liquid stops heating, be cooled to after room temperature and return to original volume with ultrapure water, making diameter is the colloid gold particle of 0.22um left and right, by 0.1M sal tartari (purchased from Tianjin chemical reagent factory) solution adjusting collaurum pH value to 6.7, add goat anti-human igg antibody (purchased from Sigma reagent company of the U.S.) to mix by 2.5mg antibody/ml collaurum, the colloid gold particle mark that is 0.22um with above-mentioned diameter by goat anti-human igg antibody (purchased from Sigma reagent company of the U.S.) obtains.
7, the packing of gram-bacteria quick detection kit (20 parts/box)
1) sample dilution 5ml/ bottle, one bottle;
2) cleansing solution 5ml/ bottle, one bottle;
3) golden labeling antibody liquid 5ml/ bottle, one bottle;
4) reaction plate, 20 parts;
5) be placed in 4 ℃ of stored refrigerated, for subsequent use.
8, detecting step operation:
1) get sample dilution in kit and splash into 3 at 1.5ml centrifuge tube, add 20ul serum specimen, mix;
2) get one of reaction plate, in reaction plate hole, add the above-mentioned sample mixing of 50ul, after specimen fluids is all permeated, splash into cleansing solution and clean 3 times, each 3.
3) splash into 3 of golden labeling antibody liquid, after it all permeates, drip 3 cleansing solutions and clean, result of determination.
9, result interpretation: by the naked eye, if purple red color spot point appears in reaction plate upper left side check point and Quality Control point, judge that bacterium infection type is as gram-positive bacteria; If purple red color spot point appears in right side check point and Quality Control point on reaction plate, judge that bacterium infection type is as Gram-negative bacteria; If Quality Control point does not develop the color, be judged to be kit invalid.
The checking of the inventive method, Gram-positive sample detects 19 examples, recall rate 95%, Gram-negative bacteria detects 18 examples, recall rate 90%; Kit can accurately detect patient bacterium infection type, highly sensitive, high specificity, and taper in 10min whole detection time.
Claims (4)
1. a method for fast detecting gram-bacteria, is characterized in that: implement step by step;
1) gram-bacteria is cultivated;
2) with nanosecond pulse, bacterium is carried out to low temperature inactivation treatment, extract gram-positive bacteria LTA antigen and Gram-negative bacteria boivin antigen;
Wherein prepare gram-positive bacteria membranous antigen: by staphylococcus aureus reference culture ATCC25923, after 37 ℃ of cultivation 12h, collect bacterium colony, after the burst process deactivation of 300ns, ultrasonic degradation, under 15000rpm condition, centrifugal 15min, obtain with CNB2 chromatographic column chromatography through cold phenol extracting again, after preparing, be stored in-80 ℃ of low temperature and preserve, for subsequent use;
Wherein prepare Gram-negative bacteria membranous antigen: by colon bacillus reference culture ATCC25922, after 37 ℃ of cultivation 12h, collect bacterium colony, after the burst process deactivation of 300ns, ultrasonic degradation, under 15000rpm condition, centrifugal 15min, through triton 114 surfactant extractings, heat to 72 ℃ again, then under 15000rpm centrifugal condition, centrifugal 15min, filter, get supernatant and cross sephadexG-200 molecular sieve purification and make, prepare, be stored in-80 ℃ of low temperature and preserve, for subsequent use;
3) by the two kinds of antigen extracting and quality controlled serum point on the nitrocellulose membrane reaction plate preparing;
On the fixed position of nitrocellulose membrane, click and enter respectively with pipettor, 2 check points of Gram-negative bacteria membranous antigen and 1 click and enter the Quality Control point of 30 normal pooled serums, point film amount 1-10ul, gram-positive bacteria membranous antigen concentration 0.7-2.2mg/ml, Gram-negative bacteria membranous antigen concentration 0.3-0.8mg/ml, after some film, 37 ℃ of baking 1h, put 4 ℃ of stored refrigerated, for subsequent use;
Wherein nitrocellulose membrane soaks 10min through 10% methyl alcohol, takes out cutting, is arranged in the hole of PVC reaction plate that is lined with adsorptive pads, after 37 ℃ of baking 2h, uses;
Wherein 2 check points and 1 the Quality Control point on nitrocellulose membrane from left to right arranged and formed according to the order of gram-positive bacteria membranous antigen check point, normal serum Quality Control point, Gram-negative bacteria membranous antigen check point successively;
4) adopt dot immunogold diafiltration technology for detection antibody, whether judgement sample contains LTA antibody and lipopolysaccharides antibody.
2. a kit for fast detecting gram-bacteria, is characterized in that: kit is made up of reaction plate, sample dilution, cleansing solution and golden labeling antibody liquid;
Reaction plate: be that reaction film is arranged in the reacting hole of the PVC base plate that is lined with adsorptive pads and is made;
Sample dilution: with 1mmol/L, Tris-HCL damping fluid 50ml, the polysorbas20 25ul of pH 8.5, gelatin 1ml, ultrapure water is settled to 100ml, makes;
Cleansing solution: with 1mmol/L, Tris-HCL damping fluid 50ml, the polysorbas20 500ul of pH 8.5, Nacl 2g, ultrapure water is settled to 100ml, makes;
Gold labeling antibody liquid: adopt trisodium citrate reduction method to prepare collaurum, colloid gold particle diameter 0.22um; Colloidal gold solution pH 6.7, light absorption value 0.08-0.15, by colloid gold label goat anti-human igg antibody, goat anti-human igg's concentration 1-4mg/ml, makes golden labeling antibody liquid.
3. according to method claimed in claim 1, it is characterized in that: the gram-positive bacteria membranous antigen of selecting is staphylococcus aureus reference culture ATCC25923 LTA antigen, molecular weight 10-100kda; The Gram-negative bacteria membranous antigen of selecting is colon bacillus reference culture ATCC25922 boivin antigen, molecular weight 10-100kda.
4. according to kit described in claim 2, it is characterized in that: the Tris-HCL that selects, polysorbas20, goat anti-human igg are all purchased from Sigma reagent company of the U.S..
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201410033894.4A CN103792351A (en) | 2014-01-23 | 2014-01-23 | Method for rapidly detecting gram bacteria as well as kit |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201410033894.4A CN103792351A (en) | 2014-01-23 | 2014-01-23 | Method for rapidly detecting gram bacteria as well as kit |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN103792351A true CN103792351A (en) | 2014-05-14 |
Family
ID=50668220
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201410033894.4A Pending CN103792351A (en) | 2014-01-23 | 2014-01-23 | Method for rapidly detecting gram bacteria as well as kit |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN103792351A (en) |
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104502589A (en) * | 2014-12-17 | 2015-04-08 | 中国科学院苏州生物医学工程技术研究所 | Chromatographic test strip for detecting platelet product bacterial pollution and detection method |
| CN105699643A (en) * | 2016-03-08 | 2016-06-22 | 湖北工业大学 | Pretreatment liquid applied before analysis of pathogenic bacterium sample and pretreatment method |
| CN105785050A (en) * | 2016-04-18 | 2016-07-20 | 石河子大学 | Comprehensive buffer solution for detecting brucellosis by means of colloidal immuno-gold immuno-filtration assay |
| CN112945914A (en) * | 2019-12-10 | 2021-06-11 | 中国科学院大连化学物理研究所 | Fluorescence detection method for rapidly distinguishing gram types of bacteria |
Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1405564A (en) * | 2001-08-17 | 2003-03-26 | 上海数康生物科技有限公司 | Reagent case for synchronous detecting multiple kinds of infectious disease and its preparing method |
| CN1409111A (en) * | 2001-09-28 | 2003-04-09 | 上海数康生物科技有限公司 | Device for simultanously detecting multiple antigen and antibody associated with diseases |
| CN101819209A (en) * | 2010-05-15 | 2010-09-01 | 厦门出入境检验检疫局检验检疫技术中心 | Method for detecting foot and mouth disease (FMD) non-structural protein 3AB antibody by using dot immuno-gold filtration assay (DIGFA) |
| CN101839908A (en) * | 2010-05-28 | 2010-09-22 | 李金波 | Quantitative detection device and detection method of biological fluid samples |
-
2014
- 2014-01-23 CN CN201410033894.4A patent/CN103792351A/en active Pending
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1405564A (en) * | 2001-08-17 | 2003-03-26 | 上海数康生物科技有限公司 | Reagent case for synchronous detecting multiple kinds of infectious disease and its preparing method |
| CN1409111A (en) * | 2001-09-28 | 2003-04-09 | 上海数康生物科技有限公司 | Device for simultanously detecting multiple antigen and antibody associated with diseases |
| CN101819209A (en) * | 2010-05-15 | 2010-09-01 | 厦门出入境检验检疫局检验检疫技术中心 | Method for detecting foot and mouth disease (FMD) non-structural protein 3AB antibody by using dot immuno-gold filtration assay (DIGFA) |
| CN101839908A (en) * | 2010-05-28 | 2010-09-22 | 李金波 | Quantitative detection device and detection method of biological fluid samples |
Non-Patent Citations (7)
| Title |
|---|
| MARTA GRANSTRöM等: "Enzeme-linked immunosorbent assay for antibodies against teichoic acid in patients with staphylococcal infections.", 《J CLIN MICROBIOL》, vol. 17, no. 4, 31 December 1983 (1983-12-31) * |
| USHA R.POTHAKAMURY等: "Inactivation of Escherichia coli and Staphylococcus aureus in model foods by pulsed electric field technology.", 《FOOD RESEARCH INTERNATIONAL》, vol. 28, no. 2, 31 December 1995 (1995-12-31) * |
| 唐秋艳 等主编: "《免疫诊断试剂实用技术》", 31 August 2009, article "胶体金免疫技术" * |
| 曹春梅: "胶体金技术在鸡白痢/鸡伤寒检疫和沙门氏菌O9抗原检测中的应用研究", 《中国优秀硕士学位论文全文数据库》, no. 200404, 15 April 2004 (2004-04-15) * |
| 曾新安，陈勇主编: "《脉冲电场非热灭菌技术》", 31 January 2005, article "脉冲电场对微生物的作用机理" * |
| 陈拯 等: "金黄色葡萄球菌感染的磷壁酸抗体检测及其意义", 《四川医学院学报》, vol. 14, no. 3, 31 December 1983 (1983-12-31) * |
| 驹形和男: "《微生物化学分类的实验方法》", 30 April 1989, article "其他细胞壁成分的分析方法" * |
Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104502589A (en) * | 2014-12-17 | 2015-04-08 | 中国科学院苏州生物医学工程技术研究所 | Chromatographic test strip for detecting platelet product bacterial pollution and detection method |
| CN105699643A (en) * | 2016-03-08 | 2016-06-22 | 湖北工业大学 | Pretreatment liquid applied before analysis of pathogenic bacterium sample and pretreatment method |
| CN105699643B (en) * | 2016-03-08 | 2017-07-21 | 湖北工业大学 | Pretreatment fluid and preprocess method before pathogen sample analysis |
| CN105785050A (en) * | 2016-04-18 | 2016-07-20 | 石河子大学 | Comprehensive buffer solution for detecting brucellosis by means of colloidal immuno-gold immuno-filtration assay |
| CN112945914A (en) * | 2019-12-10 | 2021-06-11 | 中国科学院大连化学物理研究所 | Fluorescence detection method for rapidly distinguishing gram types of bacteria |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN104662425B (en) | Vital staining method | |
| US20220236265A1 (en) | Sequential lateral flow device | |
| CN103792351A (en) | Method for rapidly detecting gram bacteria as well as kit | |
| JP5237305B2 (en) | Detection and identification of microorganisms on permeable membranes | |
| US7851207B1 (en) | Multiplex field device to detect and identify a variety of microbial agents simultaneously | |
| US20110117540A1 (en) | Highly Simplified Lateral Flow-Based Nucleic Acid Sample Preparation and Passive Fluid Flow Control | |
| Borkar | Laboratory techniques in plant bacteriology | |
| Fu et al. | Integrated paper-based detection chip with nucleic acid extraction and amplification for automatic and sensitive pathogen detection | |
| CN101509041A (en) | Chlamydia trachomatis nucleic acid detection kit for constant-temperature amplification by using RNA | |
| US3416998A (en) | Method of detecting or classifying microorganisms using agar reagent sheets | |
| CN104198704B (en) | A kind of pertussis diagnostic kit and preparation method | |
| Dezfulian et al. | Cultural and physiological characteristics and antimicrobial susceptibility of Clostridium botulinum isolates from foodborne and infant botulism cases | |
| CN110095599A (en) | The Microimmunofluorescence test method of cell-free loss | |
| CN107164497A (en) | Loop-mediated isothermal amplification technique detects the primer and kit of pseudomonas aeruginosa | |
| CN105116150A (en) | Mycoplasma pneumoniae antigen detection kit and detection method thereof | |
| CN103547922B (en) | For the method from sample detection cell | |
| CN106544436B (en) | A kind of method of salmonella in detection textile | |
| CN102243238B (en) | Nucleic acid gold-labeled rapid detection method and kit for pathogen | |
| CN110129188B (en) | Integrated nucleic acid detection device | |
| CN106323723A (en) | Double-blue staining method | |
| CN103412116B (en) | Full-automatic tubercle bacillus coloring system | |
| CN110988344A (en) | Fluorescent staining reagent for rapidly identifying staphylococcus aureus and preparation method thereof | |
| CN102816715B (en) | Kit for separating and identifying yersinia enterocolitica as well as preparation and application thereof | |
| CN101935685A (en) | Preparation method of mycobacterium tuberculosis quick color-changing drug-sensitivity medium | |
| CN101560542B (en) | Diagnostic kit for mcirocolony molecular beacon culturing mycobacterium tuberculosis, preparation method and application |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C02 | Deemed withdrawal of patent application after publication (patent law 2001) | ||
| WD01 | Invention patent application deemed withdrawn after publication |
Application publication date: 20140514 |