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A technology of 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, rapid detection, and good patient compliance.
Active Publication Date: 2015-04-29
SHENZHEN BLOT BIOTECH
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[0007] In view of this, the object of the present invention is to provide a test kit for the detection of Helicobacter pylori t...
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Embodiment 1
[0044] Example 1: Preparation of Helicobacter pylori (HP) imprinted membrane
[0045] 1. Electrophoresis separation of HP whole bacterial protein
[0046] 1. Prepare sample buffer: use a sampling gun to draw 10 mL of 0.05M pH7.4 Tris-HCL, 0.5 mL of 2-mercaptoethanol, 2 mL of glycerin, 0.5 mL of 0.1% bromophenol blue, 2 mL of 10% sodium dodecyl sulfate, Mix well and serve.
[0047] 2. Dilute the whole HP bacterial protein with sample buffer to 0.04±0.005mg / mL, after 100°C for 3 minutes, cool at 4°C for later use.
[0050] 3.2 Glue filling: Measure 30±0.5mL with a graduated cylinder, absorb 1mL of n-butanol with a pipette, add it to the top of the glue, seal it and wait for solidification.
[0051] ...
Embodiment 2
[0061] Embodiment 2: kit of the present invention
[0062] A test kit for typing and detecting Helicobacter pylori, comprising the imprinted membranes prepared in Example 1, each with a length of 98±2mm and a width of 150±5mm.
Embodiment 3
[0063] Embodiment 3: kit of the present invention
[0064] A kit for detecting Helicobacter pylori typing, comprising the blotted membrane prepared in Example 1, an enzyme-labeled antibody and an enzyme substrate, as shown in Table 1.
[0065] Table 1 Kits for Helicobacter pylori typing and detection
[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 with an application date of November 26, 2012, an application number of 201210486563.7, and an invention titled "a kit for detection of Helicobacter pylori typing". technical field [0002] The invention belongs to the field of biotechnology, and in particular relates to a test kit for typing and detecting Helicobacter pylori. Background technique [0003] Helicobacter Pylori, referred to as HP, was first isolated from gastric mucosal tissue by Australian scholars Barry Marshall and Robin Warren in 1983. Years of research by scholars from various countries have confirmed that Hp plays an important role in the pathogenesis of chronic gastritis, peptic ulcer, gastric cancer and other gastrointestinal diseases. The discovery of HP was a revolutionary development in the history of gastroenterology, and Robin Warren and Barry Marshall won the Nobel Prize in Medicine for it in 2005. [0004] The relationship between HP and gastr...
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