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Human immunodeficiency virus (HIV) nucleic acid detection kit

A technology for human immunodeficiency and detection kits, which is applied in the determination/inspection of microorganisms, microorganisms, biochemical equipment and methods, etc. To improve detection sensitivity, reduce the probability of missed detection, and achieve the effect of accurate quantification

Active Publication Date: 2013-05-01
SANSURE BIOTECH INC
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The diagnostic system has the advantages of high degree of automation, easy operation, and high detection sensitivity (greater than 40copies / ml); however, the sample requirement (1ml) is too large, and the cost of reagents and consumables for fully automated operation is too high, making it difficult to apply in clinical practice. Widely carried out

Method used

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  • Human immunodeficiency virus (HIV) nucleic acid detection kit

Examples

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Embodiment 1

[0022] This example is a total of six sets of primer probe sequences designed in the three conserved regions of LTR, GAG and POL of HIV virus. In the present invention, the primer probe design method is to download the HIV-1 sequence announced by NCBI, HIV database, etc., and select the conserved region for the gene conservative region LTR, GAG and POL gene of HIV-1; through sequence comparison, the following primers are respectively designed Probe:

[0023] LTR1:

[0024] Upstream primer LTR-F1: 5'-AGCTTGCCTTGAGTGCTTCA-3',

[0025] Downstream primer LTR-R1: 5'-AGTGGTCTGAGGGATCTCTAGTTAC-3',

[0026] Probe LTR-P1: 5'-AGTCACACAACAGACGGGCACACACT-3';

[0027] LTR2:

[0028] Upstream primer LTR-F2: 5'-CCTCAATAAAGCTTGCCTTGA-3',

[0029] Downstream primer LTR-R2: 5'-GGCGCCACTGCTAGAGATT-3',

[0030] Probe LTR-P2: 5'-ACTCTGGTAACTAGAGAGATCCCTCAGACC-3';

[0031] GAG1:

[0032] Upstream primer GAG-F1: 5'-AGCCCAGAAGTAATACCCATGTT-3',

[0033] Downstream primer GAG-R1: 5'-CATTCTGCAG...

Embodiment 2

[0049] This embodiment is the composition and operation steps of an HIV detection kit using two sets of primer probe sequences, namely LTR1+POL1.

[0050] The composition of the kit includes the following components ①~⑩,

[0051] ①RNA extraction solution Ⅰ: composed of sodium lauryl sulfate 0.2~1.0% (mass / volume), triton 1.0~4.0% (volume / volume), guanidine isothiocyanate 0.2~1.0mol / L and 100~400μg / ml of magnetic beads; this component can lyse virus particles in serum and plasma samples and release nucleic acids;

[0052] ②RNA extraction solution Ⅱ: including 4-hydroxyethylpiperazineethanesulfonic acid 100~300mmol / L, sodium chloride 100~300mmol / L, the pH value of solution Ⅱ is 6.5±0.2; it is a buffer for magnetic beads to adsorb nucleic acid solution;

[0053] ③ RNA extraction solution III: containing Triton 0.1~1.0% (vol / vol) and sodium chloride 100~300mmol / L;

[0054] ④ RNA extraction solution IV: containing mineral oil;

[0055] Wherein, RNA extraction solutions III and...

Embodiment 3~13

[0092] It is a detection kit for the other eleven kinds of primer-probe sequence combinations except that the two sets of primer-probe sequences in Example 1 are LTR1+POL1.

[0093] The components of the kit are the same as those in Example 2 except that the primer probe sequences of the target nucleotides are different (see Example 1 for the respective primer probe sequences), and the rest of the components are the same; the operation method of the kit is also the same. Consistent with the method of operation in Example 2.

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Abstract

The invention provides a human immunodeficiency virus (HIV) nucleic acid detection kit. The kit comprises at least two sets of primer probes which are respectively from different conservative regions. Preferably, the conservative regions comprise the relative conservation regions of HIV gene, i.e. GAG, POL and LTR regions. According to the invention, the two sets of designed and combined primer probe preparation reaction systems from two different regions jointly detect HIV-1RNA to identify the existence of positive reaction, effectively lower the probability of detection miss, can more comprehensively cover all subtypes, including all the subtypes of M, N and O groups of HIV-1, and further improve detection sensitivity and quantification accuracy, wherein the detection sensitivity can reach 50 IU / ml, and the linear range of the quantification is 50-1.0*108 IU / ml. The human immunodeficiency virus (HIV) nucleic acid detection kit provided by the invention further comprises a set of primer probe sequence, which is selected from any one of six sets of sequences including LTR1, LTR2, GAG1, GAG2, POL1 and POL2.

Description

technical field [0001] The invention belongs to the technical field of diagnostic reagents, in particular to a fluorescent PCR detection kit for human immunodeficiency virus HIV-1. Background technique [0002] Acquired immunodeficiency syndrome (AIDS) is a disease caused by human immunodeficiency virus (human immunodeficiency virus, HIV) infection, also known as AIDS. The immune function of patients with this disease is partially or completely lost, and the number of CD4+ cells (CD4 and CD8 are antigenic proteins on T lymphocytes, and CD4+ is called helper T cells) decreases, followed by opportunistic infections, tumors, etc., clinical manifestations Various. [0003] Since the first AIDS case was reported in the world in 1981, in just 30 years, AIDS has ravaged the world, killing more than 25 million people. UNAIDS released its annual report on the 21st. In 2010, there were about 34 million people living with HIV worldwide, an increase of 700,000 compared with 2009. Sin...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/70C12Q1/68C12R1/93
Inventor 戴立忠熊晓燕邓中平
Owner SANSURE BIOTECH INC
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