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Full-automatic rapid hepatitis B virus nucleic acid testing reagent tube and application method thereof

A hepatitis B virus and detection reagent technology, which is applied in the field of fully automatic rapid nucleic acid detection reagent tubes, can solve the problems of increasing the cost of PCR detection, showing limitations, and cumbersome procedures, etc., to eliminate the risk of false positive contamination and to operate easily OK, high sensitivity effect

Inactive Publication Date: 2014-03-12
ZHUJI MAIDA BIOTECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] However, with the popularization and application of molecular diagnostic techniques such as PCR in clinical testing, its limitations are increasingly apparent.
It is mainly manifested in two aspects. First, due to the high sensitivity of conventional PCR technology, it is very easy to produce false positive results due to trace sample contamination. Therefore, conventional PCR detection requires high quality of operators and operating environment. According to the standards of the Ministry of Health If domestic hospitals use conventional PCR technology in clinical testing items, they must establish a dedicated PCR laboratory and provide professional training for operators before proceeding, which will inevitably increase the cost of PCR testing.
Second, before performing PCR reaction detection, it is still necessary to rely on traditional manual methods to extract and purify nucleic acids in clinical samples, and then mix them with PCR reagents for detection reactions. The procedures are cumbersome, time-consuming and labor-intensive, which limits the The popularization and application of PCR detection in clinical diagnosis, especially for the detection of large batches of samples

Method used

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  • Full-automatic rapid hepatitis B virus nucleic acid testing reagent tube and application method thereof
  • Full-automatic rapid hepatitis B virus nucleic acid testing reagent tube and application method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0052] Such as figure 1 , 2 As shown, it is a fully automatic rapid nucleic acid detection reagent tube for hepatitis B virus of the present invention, including a serum sample cavity 1 for loading a serum sample to be tested and a detection reagent cavity, and the detection reagent cavity includes:

[0053] Lysis buffer chamber 2 for filling the lysis buffer,

[0054] Nucleic acid extraction magnetic bead cavity 4 for loading nucleic acid extraction magnetic beads,

[0055] Nucleic acid extraction washing liquid cavity 3 for loading nucleic acid extraction washing liquid,

[0056] Nucleic acid extraction eluent cavity 5 for loading nucleic acid extraction eluent,

[0057] Fluorescent quantitative PCR reaction solution cavity 6 for loading fluorescent quantitative PCR reaction solution,

[0058] And PCR reaction primers and probe chamber 7 for loading PCR reaction primers and probes;

[0059] The sequence of the PCR reaction primer is:

[0060] HBV1:5'-CGCACCTCTCTTTACG-3...

Embodiment 2

[0081] The fully automatic rapid nucleic acid detection reagent tube for hepatitis B virus in Example 1 of the present invention was used to detect the plasmid standard containing hepatitis B virus DNA.

[0082] Add 50 μL of Escherichia coli liquid containing hepatitis B virus DNA plasmid standard to the fully automatic rapid nucleic acid detection reagent tube of hepatitis B virus in Example 1 of the present invention, and then put the detection reagent tube into a tube type fully automatic nucleic acid Extract the amplification instrument, set the reaction program according to the method used in Example 1, lyse for 30 seconds; magnetic beads bind for 30 seconds; wash for 40 seconds; Carry out 40 variable temperature cycles, a single variable temperature cycle includes 95° C. for 4 seconds and 60° C. for 8 seconds, and the entire reaction time is about 20 minutes to complete.

[0083] The detection data of Example 2 were compared with the following comparative experiments 1 a...

Embodiment 3

[0089] The fully automatic rapid nucleic acid detection reagent tube for hepatitis B virus in Example 1 of the present invention was used for the detection of DNA in serum samples of clinical hepatitis B patients.

[0090] 100 μL containing HBV 2 x 10 5 The copies / mL clinical serum sample is added to the fully automatic rapid nucleic acid detection reagent tube of hepatitis B virus in the embodiment 1 of the present invention, and then the detection reagent tube is put into a tube type automatic nucleic acid extraction and amplification instrument, according to the embodiment 1 How to use Set the reaction program, cleavage for 30 seconds; magnetic beads binding for 30 seconds; wash for 40 seconds; 95°C for 4 seconds and 60°C for 8 seconds, the total reaction time is about 20 minutes to complete.

[0091] The detection data of Example 3 was compared with the following comparative experiments 3 and 4. Each experiment was repeated three times, and the Ct value detection data of...

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Abstract

The invention discloses a full-automatic rapid hepatitis B virus (HBV) nucleic acid testing reagent tube which is simple to operate, rapid to react and high in sensitivity. The invention further discloses an application method of the full-automatic rapid hepatitis B virus (HBV) nucleic acid testing reagent tube. According to the reagent tube, a straight-through type quantitative PCR (Polymerase Chain Reaction) technique is adopted, automatic nucleic acid extraction mainly based on nano magnetic beads is organically combined with fluorogenic quantitative PCR reaction, and full-sealed full-automatic rapid detection on hepatitis B virus (HBV) nucleic acid is realized, and the reagent tube is simple, convenient and easy to operate, rapid in reaction and high in sensitivity, the operation environment is relatively not strict, no special PCR laboratory is needed to be constructed, and labor and materials are saved.

Description

technical field [0001] The invention relates to a fully automatic rapid nucleic acid detection reagent tube for hepatitis B virus (HBV), and also relates to a method for using the fully automatic rapid nucleic acid detection reagent tube for hepatitis B virus (HBV). Background technique [0002] Hepatitis B virus (HBV) belongs to the hepadnaviridae family and mainly invades liver cells, colonizes and replicates in liver cells and damages liver cells, causing inflammation, necrosis, and fibrosis of liver cells. The current hepatitis B virus carrier rate in my country is 7.18%, and about one-third of them have recurrent liver damage, manifested as active hepatitis B or cirrhosis. The hepatitis B virus genome is about 3.2kb in length and is a partially double-stranded circular DNA with four open reading frames encoding the core nucleocapsid protein and S envelope protein of the virus, the viral replicase (polymerase) and a Protein X involved in viral gene expression. [0003]...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/70C12Q1/68
CPCC12Q1/70C12Q1/686
Inventor 夏放
Owner ZHUJI MAIDA BIOTECH
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