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A kit for detecting hepatitis B virus and a detection method for hepatitis B virus

A hepatitis B virus and kit technology, applied in biochemical equipment and methods, microbial measurement/inspection, etc., can solve problems such as time-consuming, difficult detection, false positives, etc., and achieve improved detection sensitivity, high sensitivity, and low cost Effect

Active Publication Date: 2021-06-04
NANJING AGRICULTURAL UNIVERSITY +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Therefore, these detection methods require laboratory-specific equipment and professional experimenters, take a long time, and sometimes produce false positive results, which brings certain difficulties to the detection

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  • A kit for detecting hepatitis B virus and a detection method for hepatitis B virus
  • A kit for detecting hepatitis B virus and a detection method for hepatitis B virus
  • A kit for detecting hepatitis B virus and a detection method for hepatitis B virus

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preparation example Construction

[0029] In the present invention, the liposome-QD complex is carboxyl functionalized liposome quantum dot microspheres. The preparation method of the liposome-QD complex comprises: mixing distearoyl phosphatidylcholine, phospholipid-polyethylene glycol-carboxyl, cholesterol and CdSe quantum dots in chloroform, and preparing lipid by film hydration method. Plastid-QD complexes. Described distearoyl phosphatidylcholine, phospholipid-polyethylene glycol-carboxyl and cholesterol are the membrane material of making liposome, described distearoyl phosphatidylcholine, phospholipid-polyethylene glycol-carboxyl, The molar ratio of cholesterol to the quantum dots is 9-12:2-4:0.8-1.5:4-6.5, preferably 10-11:2.5-3.5:0.9-1.2:4.5-6. The mixing temperature is preferably 35-45°C, more preferably 38-42°C. In the present invention, the emission wavelength of the CdSe quantum dots is 488nm, 561nm or 647nm. Different emission wavelengths can be excited with the same excitation light, observed i...

Embodiment 1

[0045] A test kit for detecting hepatitis B virus, comprising: a liposome-QD complex-labeled reporter probe, a magnetic bead-modified capture probe and a buffer; the reporter probe nucleic acid sequence and the capture probe nucleic acid sequence Complementary to HBV DNA respectively. Its preparation method is as follows:

[0046] 1. Preparation of liposomal quantum dot microspheres:

[0047] Add 5mL chloroform to the long-necked flask, then add 0.1M distearoyl phosphatidylcholine 100μl, 0.1M phospholipid-polyethylene glycol-carboxyl 36ul, 50mM cholesterol 22ul, 58ul, 0.1M CdSe quantum dots with an emission wavelength of 488nm, Mix well at a temperature of 40°C. The chloroform was evaporated under reduced pressure with a rotary evaporator, and when a film appeared on the inner wall of the long-necked flask, it was dried with argon. Then add 2 mL of LPBS (pH7.4), shake and dissolve the film at 30°C, and filter the dissolved solution three times with a 0.2 μm polycarbonate me...

Embodiment 2

[0055] A test kit for detecting hepatitis B virus, comprising: a liposome-QD complex-labeled reporter probe, a magnetic bead-modified capture probe and a buffer; the reporter probe nucleic acid sequence and the capture probe nucleic acid sequence Complementary to HBV DNA respectively. Its preparation method is as follows:

[0056] 1. Preparation of liposomal quantum dot microspheres:

[0057] Add 5mL chloroform to the long-necked flask, then add 0.1M distearoylphosphatidylcholine 100μl, 0.1M phospholipid-polyethylene glycol-carboxylate 36ul, 50mM cholesterol 22ul, 58ul, 0.1M CdSe quantum dots with an emission wavelength of 561nm, Mix well at a temperature of 40°C. The chloroform was evaporated under reduced pressure with a rotary evaporator, and when a film appeared on the inner wall of the long-necked flask, it was dried with argon. Then add 2 mL of LPBS (pH7.4), shake and dissolve the film at 30°C, and filter the dissolved solution three times with a 0.2 μm polycarbonate ...

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Abstract

The invention relates to the technical field of hepatitis B virus detection, in particular to a kit for detecting hepatitis B virus and a detection method thereof. The test kit of the present invention comprises: a liposome-QD complex-labeled reporter probe, a capture probe modified by magnetic beads and a buffer; DNA is complementary. In the invention, the liposome is used to wrap the quantum dot, so that the fluorescent signal is more amplified and concentrated, which is beneficial to the detection sensitivity. The kit of the invention has high detection sensitivity and good specificity, the detection limit is 10 fmol, and the detection result can be obtained within one hour.

Description

technical field [0001] The invention relates to the technical field of hepatitis B virus detection, in particular to a method for quantitatively detecting hepatitis B virus by using liposome quantum dots. Background technique [0002] Hepatitis B virus (HBV) is a partially double-stranded DNA virus that is prevalent widely. It is the pathogenic factor that causes acute and chronic hepatitis and hepatocellular carcinoma, and seriously endangers the safety of human public health. Patients with chronic HBV infection do not show any clinical symptoms, but are highly contagious and may develop serious diseases such as liver cirrhosis and liver cancer. Therefore, detection and screening of HBV infection, evaluation of treatment effect after infection and monitoring of immune level are arduous projects. [0003] At present, the methods for monitoring hepatitis B virus in hospitals and laboratories are mainly fluorescent quantitative PCR and detection of HBsAg positive as a sign of...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/6816C12Q1/70
CPCC12Q1/6816C12Q1/706
Inventor 刘斐吴旭平李嘉豪
Owner NANJING AGRICULTURAL UNIVERSITY