CN110592180A - Urease biochemical identification tube containing lysozyme - Google Patents
Urease biochemical identification tube containing lysozyme Download PDFInfo
- Publication number
- CN110592180A CN110592180A CN201910946416.5A CN201910946416A CN110592180A CN 110592180 A CN110592180 A CN 110592180A CN 201910946416 A CN201910946416 A CN 201910946416A CN 110592180 A CN110592180 A CN 110592180A
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- CN
- China
- Prior art keywords
- urease
- biochemical
- lysozyme
- bacteria
- identification
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- 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.)
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Classifications
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
- C12Q1/02—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving viable microorganisms
- C12Q1/04—Determining presence or kind of microorganism; Use of selective media for testing antibiotics or bacteriocides; Compositions containing a chemical indicator therefor
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
- C12Q1/34—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving hydrolase
Abstract
The invention discloses a urease biochemical identification tube containing lysozyme, belonging to a method for testing materials by testing the color change of reaction results by utilizing visible light. The technical scheme of the invention is as follows: the bacteria biochemical identification tube contains lysozyme, the cell wall of the bacteria is dissolved through the action of the lysozyme, intracellular enzymes of the cells are dissociated, biochemical enzymatic reaction is carried out, and bacteriological identification is carried out according to the characteristics of the enzymes contained in each kind of bacteria. The method of the invention has the characteristics of fast dissociation of the bacterial intracellular enzyme, full reaction of the bacterial intracellular enzyme, shortened identification time, and multiple and accurate identification types of the intracellular enzyme.
Description
Technical Field
The present invention pertains to a method of assay comprising an enzyme; or a method for testing materials by using visible light and generating color change through the result of test reaction, in particular to a determination method for performing bacterial biochemical identification by using lysozyme to dissociate thallus.
Background
Bacteria identification is generally carried out by adopting bacteria plate streaking for separation, then proliferation is carried out on a enrichment medium, classification of positive bacteria, negative bacteria, cocci, bacilli and the like is carried out through gram staining, then bacteria biochemical reaction identification is carried out, and bacteria genus identification is carried out according to the special biochemical reaction and the gram staining and the shape of the bacteria. However, bacteria contain both intracellular and extracellular enzymes such as (1) plasma Coagulase (coagulose): most pathogenic staphylococcus aureus can produce a plasma coagulase (free plasma coagulase), can accelerate the coagulation of human or rabbit plasma, and can protect pathogenic bacteria from phagocytosis or antibody and the like. (2) Streptokinase (Streptokinase): or Streptococcal fibrinolysin (Streptococcal fibrinolysin), most streptococci causing human infection produce streptokinase. The effect is to activate Plasminogen or plasmogen (Plasminogen) to become fibrinolytic enzyme or a cytotoxic enzyme (Plasmin) to dissolve the fibrin clot. (3) Hyaluronidase (Hyaluronidase): or Spreading factor (enzyme), which can dissolve hyaluronic acid in connective tissue of organism, make connective tissue loose and increase permeability. The biochemical identification of bacteria is carried out by the action of extracellular and secreted enzymes or by the biochemical reaction of bacteria that have been lysed, which often has limitations in the biochemical identification of bacteria.
Lysozyme (also called muramidase) or N-acetylmuramidase hydrolase (N-acetylmuramidase glycohydrolase) is an alkaline enzyme that hydrolyzes mucopolysaccharides in pathogenic bacteria. The bacterial lysis is achieved by breaking the beta-1, 4 glycosidic bond between N-acetylmuramic acid and N-acetylglucosamine in the cell wall, breaking down the cell wall insoluble mucopolysaccharide into soluble glycopeptides, causing the contents of the broken cell wall to escape. Lysozyme can efficiently hydrolyze peptidoglycan of bacterial cell walls at the beta-1.4 glycosidic bond between the carbon atom at position 1 of N-acetylmuramic acid (NAM) and the carbon atom at position 4 of N-acetylglucosamine (NAG). Any chemical bond breakage in the above structure, which serves as the backbone of the cell wall, can lead to damage of the bacterial cell wall.
The invention content is as follows:
in order to simply, conveniently, quickly and accurately carry out bacteria biochemical identification, the invention provides the bacteria biochemical identification tube which is economic, convenient, quick, high in accuracy and contains lysozyme.
The technical scheme adopted by the invention is as follows: the bacteria biochemical identification tube contains lysozyme, the bacteria cell wall is dissolved through the action of the lysozyme, the bacteria intracellular enzyme is dissociated out, the bacteria biochemical enzymatic reaction is carried out, and the bacteriological identification is carried out according to the characteristics of the enzyme contained by each kind of bacteria. The method of the invention has the characteristics of shortening the identification time and having multiple and accurate enzyme identification types because the dissociation of the bacterial intracellular enzyme is fast and the bacterial intracellular enzyme can fully react.
The lysozyme component is contained in all bacteria biochemical identification tubes, and the concentration of the lysozyme component is within the protection scope of the invention.
The specific implementation mode is as follows:
the present invention will be described in further detail below with reference to examples of urease-bacteria-identifying tubes.
Example 1
The composition of the reagent is as follows:
urease bacterium identification tube: 100ml of broth contains 500U of lysozyme, 5mg of urea and 40 mul of phenol red, and the urease-producing bacteria can decompose urea to produce ammonia so that the broth becomes alkaline. The indicator is phenol red, the positive is red, and the negative is orange. The measuring tube can be made into a 2ml ampoule for standby and is a disposable consumable.
Example 2
Taking an Escherichia coli standard strain ATCC 25922 as an example, the urease bacterium identification tube containing lysozyme and a commercially available lysozyme-free bacterium identification tube are respectively used for carrying out biochemical reaction tests, and the test results show that the test result of the invention is positive within 3 hours, while the commercially available lysozyme-free bacterium identification tube is obviously positive within 22 hours, and the strength of the positive result is obviously lower than that of the urease bacterium identification tube containing lysozyme.
The experiment comparison shows that the sensitivity and the reaction time of the invention are obviously better than those of the original commercial urease bacterium identification tube.
Claims (4)
1. A urease biochemical identifying tube containing lysozyme features that the biochemical identifying tube contains lysozyme, the lysozyme is used to dissolve the bacterial cell wall, the intracellular enzyme is dissociated out for biochemical enzymological reaction, and the bacteriological identification is performed according to the characteristics of the enzyme contained in each bacterium.
2. The urease biochemical identification tube containing lysozyme according to claim 1 wherein the urease bacteria identification tube contains 50 to 5000U lysozyme, 0.5 to 50mg urea and 0.5 to 400 μ l phenol red in 100ml broth culture medium, and the urease bacteria can decompose urea to produce ammonia to cause the culture medium to change alkali and develop color.
3. The urease biochemical identification tube according to claim 1 wherein the indicator is phenol red, and the urease-producing bacteria can decompose urea to produce ammonia and the medium becomes alkaline, red positive and orange negative.
4. The urease biochemical assay tube containing lysozyme according to claim 1, wherein the assay tube is a disposable consumable material and can be made into 2ml ampoules for later use.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201910946416.5A CN110592180A (en) | 2019-10-05 | 2019-10-05 | Urease biochemical identification tube containing lysozyme |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201910946416.5A CN110592180A (en) | 2019-10-05 | 2019-10-05 | Urease biochemical identification tube containing lysozyme |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN110592180A true CN110592180A (en) | 2019-12-20 |
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Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201910946416.5A Pending CN110592180A (en) | 2019-10-05 | 2019-10-05 | Urease biochemical identification tube containing lysozyme |
Country Status (1)
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Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| EP0018825A1 (en) * | 1979-05-02 | 1980-11-12 | National Research Development Corporation | A process for the identification of bacteria and a kit of reagents for use in this process |
| US5935804A (en) * | 1997-03-21 | 1999-08-10 | Laine; Roger A. | Method for detecting eubacteria in biological samples with catalytically inactive murein binding enzymes |
| US20110076716A1 (en) * | 2009-09-28 | 2011-03-31 | Sysmex Corporation | Bacteria analysis apparatus, bacteria analysis method and computer program product |
| TH112259A (en) * | 2007-06-01 | 2012-02-20 | นางสาวอรุณศรี ศรีธนะอิทธิพล | "Rapid enzyme detection and screening process" |
| US20180128828A1 (en) * | 2016-11-04 | 2018-05-10 | James W. Stave | Direct detection of microorganisms in patient samples by immunoassay |
| CN209412237U (en) * | 2018-06-04 | 2019-09-20 | 上海市食品药品检验所 | A kind of bacterial identification kits based on DNA characteristics sequence |
-
2019
- 2019-10-05 CN CN201910946416.5A patent/CN110592180A/en active Pending
Patent Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| EP0018825A1 (en) * | 1979-05-02 | 1980-11-12 | National Research Development Corporation | A process for the identification of bacteria and a kit of reagents for use in this process |
| US5935804A (en) * | 1997-03-21 | 1999-08-10 | Laine; Roger A. | Method for detecting eubacteria in biological samples with catalytically inactive murein binding enzymes |
| TH112259A (en) * | 2007-06-01 | 2012-02-20 | นางสาวอรุณศรี ศรีธนะอิทธิพล | "Rapid enzyme detection and screening process" |
| US20110076716A1 (en) * | 2009-09-28 | 2011-03-31 | Sysmex Corporation | Bacteria analysis apparatus, bacteria analysis method and computer program product |
| US20180128828A1 (en) * | 2016-11-04 | 2018-05-10 | James W. Stave | Direct detection of microorganisms in patient samples by immunoassay |
| CN209412237U (en) * | 2018-06-04 | 2019-09-20 | 上海市食品药品检验所 | A kind of bacterial identification kits based on DNA characteristics sequence |
Non-Patent Citations (2)
| Title |
|---|
| 朱奇等: "溶菌酶及其应用", 《生物学通报》 * |
| 潘若男等: "尿素酶试验的最佳pH", 《微生物学通报》 * |
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| PB01 | Publication | ||
| PB01 | Publication | ||
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| RJ01 | Rejection of invention patent application after publication |
Application publication date: 20191220 |
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| RJ01 | Rejection of invention patent application after publication |