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PCR primer set, probe, kit and detection method for detecting hiv-1 2-LTR DNA

A technology for detecting HIV-12-LTR DNA and primers, which is applied in biochemical equipment and methods, DNA/RNA fragments, recombinant DNA technology, etc., can solve the problems that detection methods cannot be realized, and achieve improved primer annealing temperature, high sensitivity and specific effect

Active Publication Date: 2020-07-31
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 set, probe, kit and detection method for detecting hiv-1 2-LTR DNA
  • PCR primer set, probe, kit and detection method for detecting hiv-1 2-LTR DNA
  • PCR primer set, probe, kit and detection method for detecting hiv-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'-CAAGGCTACTTCCCCTGATTA-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 a FAM fluorescent group, and the 3' end of the nucleotide sequence of probe 1 is marked with a TAMRA quencher group.

Embodiment 2

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

[0073] Detection primer sets include:

[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'-CAAGGCTACTTCCCCTGATTA-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 marked with a HEX fluorescent group, and the 3' end of the nucleotide sequence of probe 2 is marked with a quencher Group MGB.

Embodiment 3

[0082] Embodiment 3 detects the detection primer set and detection probe of human immunodeficiency virus type 1 2-LTR DNA

[0083] Detection primer sets include:

[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'-CAAGGCTACTTCCCCTGATTA-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 marked with a FAM fluorescent group, and the 3' end of the nucleotide sequence of probe 1 is marked with a TAMRA quencher killing group;

[0092] and probe 2: its sequence is 5'-ACACTACTTGAAGCACTCAAGGCAAGC-3', the 5' end of the nucleotide sequence of probe 2 is labeled with a FAM fluorescent group, and the 3' end of the nucleotide sequence of probe 2 is labeled wit...

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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, specifically relates to the field of virus detection, and more specifically relates to a PCR primer set, a probe, a kit thereof and a detection method for detecting human immunodeficiency virus type 1 2-LTR DNA. Background technique [0002] Human Immunodeficiency Virus (HIV) is a lentivirus that infects cells of the human immune system and is a type of retrovirus. Human immunodeficiency virus was first identified in the United States in 1983. The virus destroys the immune ability of the human body, causing the immune system to lose its resistance, which leads to the survival of various diseases and cancers in the human body, and eventually leads to AIDS. According to incomplete statistics, it is estimated that 35.9 to 44.3 million people in the world live with HIV, of which 4.3 to 6.4 million are new infections, and 2.8 to 3.5 million die from AIDS. [0003] In recent years, combined antiretrovira...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/70C12Q1/6851C12N15/11C12R1/93
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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