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Blood screening three-item multi-color single tube detection kit

A detection kit and single-tube technology, which is applied in the field of three multi-color single-tube detection kits for blood screening, can solve the problems of low detection sensitivity, long detection time, and poor specificity, so as to improve sensitivity, specificity, and reduce The effect of intensity

Inactive Publication Date: 2017-10-24
GUANGZHOU HEAS BIOTECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] In order to solve the problems of low detection sensitivity, poor specificity, long detection time and cumbersome process of ordinary HBV, HCV and HIV virus detection kits on the market

Method used

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  • Blood screening three-item multi-color single tube detection kit
  • Blood screening three-item multi-color single tube detection kit
  • Blood screening three-item multi-color single tube detection kit

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0035] Example 1: Operation steps of a three-item multicolor single-tube detection kit for blood screening

[0036] 1) Nucleic acid sample extraction:

[0037] It is used in conjunction with Guangzhou Heshi biological nucleic acid extraction reagent (magnetic bead method) extraction reagent, and is operated in strict accordance with the instructions for automatic nucleic acid extraction.

[0038] 2) Preparation of positive quality control products:

[0039] Pseudoviruses containing specific amplification fragments of HIV, HBV and HCV were respectively constructed as positive quality control products, and the reference for the construction method was "Construction and Expression of Ribonuclease-resistant Virus-Like Particles Containing Long Fragment Chimeric RNA (2008)" conduct;

[0040] The negative quality control substances of HIV, HBV and HCV are all H 2 O.

[0041] 3) Prepare PCR reaction system:

[0042] Put 3U Fast Taq enzyme, 3U reverse transcriptase, 12mM Tris-HCL...

Embodiment 2

[0059] Embodiment 2: three kinds of virus-specific primer probe sequence screening processes in the present invention

[0060] 1) Nucleic acid sample extraction:

[0061] It is used in conjunction with Guangzhou Heshi biological nucleic acid extraction reagent (magnetic bead method) extraction reagent, and is operated in strict accordance with the instructions for automatic nucleic acid extraction.

[0062] 2) Preparation of positive quality control products:

[0063] Pseudoviruses containing specific amplification fragments of HIV, HBV and HCV were respectively constructed as positive quality control products, and the reference for the construction method was "Construction and Expression of Ribonuclease-resistant Virus-Like Particles Containing Long Fragment Chimeric RNA (2008)" conduct;

[0064] The negative quality control substances of HIV, HBV and HCV are all H 2 O.

[0065] 3) Prepare PCR reaction system:

[0066] Put 3U Fast Taq enzyme, 3U reverse transcriptase, 12...

Embodiment 3

[0078] Example 3: Comparison of detection sensitivity between the MGB probe system used in the present invention and the common probe system.

[0079] 1) Preparation of positive quality control:

[0080] Pseudoviruses containing specific amplification fragments of HIV, HBV and HCV were respectively constructed as positive quality control products, and the reference for the construction method was "Construction and Expression of Ribonuclease-resistant Virus-Like Particles Containing Long Fragment Chimeric RNA (2008)" Carried out, the nucleic acid sequence of the constructed pseudovirus was verified by Sanger sequencing, and could be recognized by the aforementioned three primers and probes;

[0081] The negative quality control substances of HIV, HBV and HCV are all H 2 O;

[0082] Take the three pseudoviruses constructed above and carry out concentration calibration respectively, so that the concentration of each pseudovirus contained in the mixture is 1×10 5 IU / ml, divided...

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Abstract

The invention discloses a blood screening three-item multi-color single tube detection kit. A blood screening three-item multi-color single tube detection reagent method comprises the steps of (1) using matched Guangzhou and real biological nucleic acid extraction reagents (a paramagnetic particle method), strictly according to the specification and carrying out automatic nucleic acid extraction; (2) designing a specific primer and a corresponding MGB fluorescent probe for an HBV, an HCV and an HIV (blood screening three items); (3) mixing the specific primer, the MGB probe and a fast enzyme system for detecting blood screening three items and carrying out amplification; and (4) detecting a fluorescent change of a reaction system and determining whether HBV, HCV and HIV infection exists or not. The method is simple in operation, and is capable of quickly detecting whether HBV, HCV and HIV infection exists or not at the same time. The detection method is multi-color single tube detection, three items of the HBV, the HCV and the HIV can be detected in PCR reaction of a same tube at the same time, and the three-item detection process is greatly simplified. The probe used for the detection method is the MGB probe, so that the detection sensitivity and specificity are greatly improved. The fast enzyme system is also applied to the detection method, so that the time required for reaction is greatly shortened and the work efficiency is improved.

Description

technical field [0001] The invention relates to a three-item multicolor single-tube detection kit for blood screening, in particular to a detection kit for HBV, HCV and HIV viruses. Background technique [0002] Hepatitis B virus is called hepatitis B virus for short. Hepatitis B virus DNA is the deoxyribonucleic acid of hepatitis B virus, which is the core substance of hepatitis B virus and the basis of virus replication. In the genetic gene carried by the virus DNA, the hepatitis B virus DNA replicates a new virus shell and DNA core respectively, forming a large number of new viruses, which are released and continue to infect other liver cells, causing hepatitis B virus disease. Hepatitis C is caused by hepatitis C virus (HCV) infection. According to the World Health Organization, 170 million people worldwide are infected with HCV. The anti-HCV positive rate of healthy people in my country is 0.7%~3.1%, about 38 million people. Due to various factors such as the biologi...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/70C12Q1/68C12N15/11C12R1/93
CPCC12Q1/686C12Q1/703C12Q1/706C12Q1/707C12Q2521/101C12Q2563/107
Inventor 陈华云丁渭刘淑园陆同山
Owner GUANGZHOU HEAS BIOTECH CO LTD
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