阅读说明：本技术 含有溶菌酶的尿素酶生化鉴定管 (Urease biochemical identification tube containing lysozyme ) 是由 李立和 刘冰 魏志斌 于 2019-10-05 设计创作，主要内容包括：本发明公开了一种含有溶菌酶的尿素酶生化鉴定管,属于利用可见光,通过测试反应结果颜色的变化来测试材料的方法。本发明的技术方案是：细菌生化鉴定管中含有溶菌酶,通过溶菌酶作用使细菌细胞壁溶解,细胞胞内酶解离出来,进行生化酶学反应,根据每种细菌含有的酶特性进行细菌学鉴定。本发明方法使细菌胞内酶解离快,细菌胞内酶能够充分反应,缩短了鉴定时间,胞内酶鉴定种类多,准确的特点。(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.)

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.

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.