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A kind of test kit for typing detection of Helicobacter pylori

A technology for Helicobacter pylori and bacterial protein, which is applied in measurement devices, analytical materials, instruments, etc., can solve problems such as HP typing and typing, and achieve the effects of small sample demand, high sensitivity and wide application range.

Active Publication Date: 2016-08-17
SHENZHEN BLOT BIOTECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0007] In view of this, the object of the present invention is to provide a test kit for the detection of Helicobacter pylori typing in view of the defect that the detection of HP in the prior art can only detect whether there is HP infection but cannot type HP

Method used

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  • A kind of test kit for typing detection of Helicobacter pylori
  • A kind of test kit for typing detection of Helicobacter pylori
  • A kind of test kit for typing detection of Helicobacter pylori

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0044] Example 1: Preparation of Helicobacter pylori (HP) blotting membrane

[0045] 1. HP whole cell protein electrophoresis separation

[0046] 1. Configure sample buffer: Use a sample gun to draw 10 mL of 0.05M pH7.4 Tris-HCL, 0.5 mL of 2-mercaptoethanol, 2 mL of glycerol, 0.5 mL of 0.1% bromophenol blue, and 2 mL of 10% sodium lauryl sulfate. Mix well and get it.

[0047] 2. Dilute HP whole cell protein with sample buffer to 0.04±0.005mg / mL, treat it at 100°C for 3 minutes, and cool it at 4°C for later use.

[0048] 3. Gradient discontinuous electrophoresis

[0049] 3.1. Preparation of separation gel: measure 40% acrylamide 4mL, 40% methylene-bisacrylamide 0.1mL, pH9.2 0.5M Tris-HCL 5mL, DW 5mL, TEMED (tetramethylethylenediamine) 50μL, 1 % Ammonium persulfate 5mL, H 2 O 12.5mL.

[0050] 3.2 Glue filling: Measure 30±0.5mL with a graduated cylinder, pipette 1mL of n-butanol, add it to the glue and seal it for solidification.

[0051] 3.3 After the separation gel has solidified (20±2 m...

Embodiment 2

[0061] Example 2: The kit of the present invention

[0062] A kit for typing and detecting Helicobacter pylori includes the imprinted membrane prepared in Example 1, each with a length of 98±2mm and a width of 150±5mm.

Embodiment 3

[0063] Example 3: The kit of the present invention

[0064] A kit for typing detection of Helicobacter pylori includes the blotting membrane prepared in Example 1, an enzyme-labeled antibody and an enzyme substrate, as shown in Table 1.

[0065] Table 1 Kits for typing detection of Helicobacter pylori

[0066]

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Abstract

The invention belongs to the field of biotechnology, and discloses a kit for typing detection of helicobacter pylori. The kit comprises a blotting membrane transferred with helicobacter pylori whole cell lysate protein obtaind by electrophoretic separation. Various Hp antibodies including pathogenic factor cytotoxicity (CagA), vacuole (VacA) and urease subunit A and antibody B contained in the serum of the helicobacter pylori infected patients can be detected by adopting an immunoblotting method, whether infected HP produces strain can be determined according to different HP antibodies so as to further judge the serology type of the infected Hp strain of the patients. According to the kit, patients to be detected only needs to provide serum, tests belong to non-invasive examination, the compliance of the patients is good, the detection is fast as the whole time only lasts for 2 hours, the sample size is small as only 10mu L of to-be-detected serum is needed, the specificity is strong, the sensitivity is high, the application range is wide, and the kit is convenient for popularization and use in hospitals and hygiene departments at all levels.

Description

[0001] This application is a divisional application whose application date is November 26, 2012, the application number is 201210486563.7, and the invention is titled "a kit for typing detection of Helicobacter pylori". Technical field [0002] The invention belongs to the field of biotechnology, and specifically relates to a kit for typing and detecting Helicobacter pylori. Background technique [0003] Helicobacter Pylori, referred to as HP, was first isolated from gastric mucosal tissue in 1983 by Australian scholars Barry Marshall and Robin Warren. After years of research by scholars from various countries, it has been confirmed that Hp plays an important role in the pathogenesis of digestive tract diseases such as chronic gastritis, peptic ulcer and gastric cancer. The discovery of HP is a revolutionary development in the history of gastroenterology. In 2005, Robin Warren and Barry Marshall won the Nobel Prize in Medicine. [0004] The relationship between HP and gastroduodena...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N33/569G01N33/544
CPCG01N33/544G01N33/56911
Inventor 马伟民张永顶马新民
Owner SHENZHEN BLOT BIOTECH
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