PCR primer group, probe and kit and detection method for detecting HIV (human immunodeficiency virus)-1 2-LTR DNA

A technology for HIV-12-LTR DNA, detection primers, applied in biochemical equipment and methods, DNA/RNA fragments, recombinant DNA technology, etc. specific effect

Active Publication Date: 2017-09-01
GUANGZHOU SUPBIO BIO TECH & SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

That is to say, under the condition that the plasma virus level is lower than the detection limit, trying to describe the degree of HIV-1 virus replication under the condition of effective suppression and determine an accurate dynamic process cannot be achieved with existing detection methods

Method used

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  • PCR primer group, probe and kit and detection method for detecting HIV (human immunodeficiency virus)-1 2-LTR DNA
  • PCR primer group, probe and kit and detection method for detecting HIV (human immunodeficiency virus)-1 2-LTR DNA
  • PCR primer group, probe and kit and detection method for detecting HIV (human immunodeficiency virus)-1 2-LTR DNA

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0063] Example 1 Detection primer set and detection probe for detecting human immunodeficiency virus type 1 2-LTR DNA

[0064] The detection primer set includes: primer 1: 5'-TGGCTAACTAGGGAACCCACT-3';

[0065] Primer 2: 5'-AGACAAGATATCCTTGATCTGT-3';

[0066] Primer 3: 5’-AACTAGGGAACCCACTGCTTAAGCC-3’;

[0067] Primer 4: 5'-CCACAGATCAAGGATATCTTGTC-3';

[0068] Primer 5: 5'-CTCAGATCCTGCATATAAGC-3';

[0069] And primer 6: 5'-CAAGGCTACTTCCCTGATTA-3'.

[0070] Detection probes include:

[0071] Probe 1: Its sequence is 5'-AAGCCTCAATAAAGCTTGCCTTGAGTGC-3', the 5'end of the nucleotide sequence of probe 1 is labeled with FAM fluorophore, and the 3'end of the nucleotide sequence of probe 1 is labeled with TAMRA quenching Group.

Embodiment 2

[0072] Example 2 Detection primer set and detection probe for detecting human immunodeficiency virus type 1 2-LTR DNA

[0073] The detection primer set includes:

[0074] Primer 1: 5’-TGGCTAACTAGGGAACCCACT-3’;

[0075] Primer 2: 5'-AGACAAGATATCCTTGATCTGT-3';

[0076] Primer 3: 5’-AACTAGGGAACCCACTGCTTAAGCC-3’;

[0077] Primer 4: 5'-CCACAGATCAAGGATATCTTGTC-3';

[0078] Primer 5: 5'-CTCAGATCCTGCATATAAGC-3';

[0079] And primer 6: 5'-CAAGGCTACTTCCCTGATTA-3'.

[0080] Detection probes include:

[0081] Probe 2: Its sequence is 5'-ACACTACTTGAAGCACTCAAGGCAAGC-3', the 5'end of the nucleotide sequence of probe 2 is labeled with HEX fluorophore, and the 3'end of the nucleotide sequence of probe 2 is labeled with quencher The group MGB.

Embodiment 3

[0082] Example 3 Detection primer set and detection probe for detecting human immunodeficiency virus type 1 2-LTR DNA

[0083] The detection primer set includes:

[0084] Primer 1: 5’-TGGCTAACTAGGGAACCCACT-3’;

[0085] Primer 2: 5'-AGACAAGATATCCTTGATCTGT-3';

[0086] Primer 3: 5’-AACTAGGGAACCCACTGCTTAAGCC-3’;

[0087] Primer 4: 5'-CCACAGATCAAGGATATCTTGTC-3';

[0088] Primer 5: 5'-CTCAGATCCTGCATATAAGC-3';

[0089] And primer 6: 5'-CAAGGCTACTTCCCTGATTA-3'.

[0090] Detection probes include:

[0091] Probe 1: Its sequence is 5'-AAGCCTCAATAAAGCTTGCCTTGAGTGC-3', the 5'end of the nucleotide sequence of probe 1 is labeled with FAM fluorophore, and the 3'end of the nucleotide sequence of probe 1 is labeled with TAMRA quenching Kill group

[0092] And probe 2: its sequence is 5'-ACACTACTTGAAGCACTCAAGGCAAGC-3', the 5'end of the nucleotide sequence of probe 2 is labeled with FAM fluorophore, and the 3'end of the nucleotide sequence of probe 2 is labeled with quenching Kill the group TAMRA.

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Abstract

The invention belongs to the field of biotechnology, particularly relates to the field of virus detection, and more particularly relates to a PCR primer group, probe and kit and detection method for detecting human immunodeficiency virus-1 2-LTR DNA. The detection primer group provided by the invention comprises nucleotide sequences shown as SEQ ID NO: 1-6; and the detection probe comprises nucleotide sequences shown as SEQ ID NO: 7-8. The detection primer group provided by the invention is different from the conventional real-time fluorescence quantitative PCR; through introduction of 3 pairs of real-time fluorescent quantitative PCR primers, that is, 6 primers, adoption of multiple cycles, and increase of the primer annealing temperature, the PCR detection method for detecting the HIV-1 2-LTR DNA, provided by the invention, achieves extremely high sensitivity and specificity, and the minimum detection limit can reach 2 copies per million cells in each PCR; and moreover, when a quantitative cell reaction system is further added in the PCR reaction, the HIV-1 2-LTR DNA and the number of cells can be quantified at the same time in the same PCR reaction (in the conventional method, the cell count is additionally required).

Description

Technical field [0001] The invention belongs to the field of biotechnology, and specifically relates to the field of virus detection, and more specifically to a PCR primer set, probes, kits and detection methods for detecting human immunodeficiency virus type 1 2-LTR DNA. Background technique [0002] Human Immunodeficiency Virus (HIV) is a kind of Lentivirus (Lentivirus) that infects cells of the human immune system, which is a kind of retrovirus. In 1983, the human immunodeficiency virus was first discovered in the United States. The virus destroys the immune system of the human body and causes the immune system to lose resistance, which leads to various diseases and cancers to survive in the human body, and eventually leads to AIDS. According to incomplete statistics, there are an estimated 3590-44.3 million people living with human immunodeficiency virus worldwide, among which 4.3-6.4 million are new infections, and 2.8-3.5 million people have died of AIDS. [0003] In recent...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/70C12Q1/68C12N15/11C12R1/93
CPCC12Q1/6851C12Q1/703C12Q2531/113C12Q2545/113C12Q2563/107
Inventor 王法张晓玮彭春梅邓可基乐小炎张嘉李家导陈观芝林敏深林若琳石壮壮罗园香莫静嫣李海茵张新王星吴彩虹
Owner GUANGZHOU SUPBIO BIO TECH & SCI
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