PCR primer groups and probe used for detecting human immunodeficiency virus type I, kit containing PCR primer groups and probe, and detection method
A technology for human immunodeficiency and detection kits, applied in biochemical equipment and methods, microorganism-based methods, microorganisms, etc. Specific, clinically applicable effects
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Embodiment 1
[0060] Example 1: PCR primer set for detection of human immunodeficiency virus type 1 DNA
[0061] In the early stage of the present invention, a large number of PCR primers for detecting human immunodeficiency virus type 1 DNA were designed based on the principle of primer design and combined with the actual situation, and then high-sensitivity and high-specificity primer sequences and detection probes were screened out through a large number of experimental studies, and finally selected The PCR primer sets with the best effect for detecting human immunodeficiency virus type 1 DNA include 3 sets, and their nucleotide sequences are as follows:
[0062] Primer set 1:
[0063] SEQ ID NO: 1: TCTGGCTAACTAGGGAACCCACTGCT;
[0064] SEQ ID NO: 2: TGCGCGCTTCAAGCCGAGTCCTGCGT;
[0065] SEQ ID NO: 3: AGGGAACCCACTGCTTAAGCCTCAATAAAGCT;
[0066] SEQ ID NO: 4: AGCAAGCCGAGTCCTGCGTCGAGA;
[0067] SEQ ID NO: 5: AGCCTCAATAAAGCTTGCCT;
[0068] SEQ ID NO: 6: CCGCCACTGCTAGAGATTTTCCA;
[0069...
Embodiment 2
[0095] Example 2: PCR detection kit for detection of human immunodeficiency virus type 1
[0096] The PCR test kit for detection of human immunodeficiency virus type 1 includes the following components:
[0097] 1) containing at least one set of primers described in Example 1;
[0098] 2) Contains detection probes:
[0099] When the kit contains primer set 1, the kit also contains probe SEQ ID NO: 15 or its nucleotide complement;
[0100] When the kit contains primer set 2, the kit also contains at least one of the probes SEQ ID NO: 16, SEQ ID NO: 17, SEQ ID NO: 18 or the nucleotide complementary sequences of these sequences probe;
[0101] When primer set 3 is contained in the kit, the kit also contains probes SEQ ID NO: 19, SEQ ID NO: 20, SEQ ID NO: 21, SEQ ID NO: 22 or nucleotides of these sequences at least one probe in the complementary sequence.
[0102] The fluorescent group labeled at the 5' end of the probe sequence is any one of FAM, HEX, VIC, CY5, and TET...
Embodiment 3
[0114] Example 3: Detection method of human immunodeficiency virus type 1 PCR detection kit
[0115] Utilize the detection kit established in Example 2 to detect the sample to be detected, and the steps are as follows:
[0116] 1) DNA extraction:
[0117] Take 200 μl of the whole blood sample to be tested, negative quality control material, weak positive quality control material, strong positive quality control material, and positive quantitative reference material, use the QIAamp DNA Blood Mini Kit extraction kit, and follow the whole blood sample extraction steps in the instruction manual. , the elution volume is 100uL, and the DNA in each group of samples is extracted and used as the subsequent PCR template;
[0118] 2) PCR reaction system:
[0119] PCR reaction solution 29.2μL;
[0121] DNA template 20μL;
[0122] 3) PCR reaction program:
[0123] Taking ABI7500 as an example, the first step: 37°C for 5 minutes; the second step: 95°C f...
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