Apparatus for integrated real-time nucleic acid analysis, and method for detecting a target nucleic acid using same
An integrated device and nucleic acid analysis technology, which is applied in the field of real-time nucleic acid analysis integrated devices, can solve the problems of 96-well nucleic acid amplification reactor waste, economic time limit, wasteful device operation, etc., and achieve shortened test time, efficient use, and saving The effect of fees
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Embodiment 1
[0060] Embodiment 1 relates to the real-time nucleic acid analysis integrated device of the present invention, figure 1 A schematic diagram of the real-time nucleic acid analysis integrated device of the present invention is shown, and the present invention will be described in more detail below.
[0061] The integrated real-time nucleic acid analysis device includes a plurality of automatic purification and dispensation devices 100 for separating and purifying different nucleic acids from multiple types of biological samples containing nucleic acids. The above-mentioned biological sample is a biological substance isolated in a natural environment and containing nucleic acid. The biological sample of the present invention may be an isolated or purified nucleic acid solution, microorganism, tissue, biological fluid or cell. Biological fluids include, but are not limited to, blood, plasma, sputum, urine, cerebrospinal fluid, gastric washings, leukophoresis, and various specimen...
Embodiment 2
[0106] Embodiment 2 relates to the target nucleic acid detection method using the real-time nucleic acid analysis integrated device of the present invention, which will be described in more detail below.
[0107] The present invention provides a target nucleic acid detection method using a real-time nucleic acid analysis integrated device, which is characterized in that it comprises:
[0108] 1) In a plurality of automatic purification and dispensing devices 100, the nucleic acid contained in each sample is separated and purified from the standard sample and various biological samples containing nucleic acid, and then the separated and purified solution is added to the nucleic acid amplification reaction mixture The step of mixing the reaction tube;
[0109] 2) By making the porous temperature circulation block 210 in which the reaction tube of the nucleic acid amplification reaction mixture is installed correspond to each target nucleic acid separated and purified from the au...
Embodiment 3
[0159] Example 3 relates to the internal control for the separation and purification of nucleic acid and the simultaneous detection of multiple samples in the target nucleic acid detection method using the integrated real-time nucleic acid analysis device of the present invention. The present invention will be described in more detail below.
[0160] (1) Design and application of plant internal control primers and probes
[0161]Plant internal control (Internal Positive Control, IPC) is the base sequence 4903 to 5709 (Movement protein gene, MP) in the genome RNA (GeneBank.ACCESSION No.NC001367) of tobacco mosaic virus (Tobacco mosaic virus, TMV), Between 5712 and 6191 (Coat protein gene, CP), base sequences with a length of 18-28 bp and a Tm value of 55° C. to 62° C. were randomly selected as forward and reverse primers.
[0162] In addition, among the above-mentioned base sequences, base sequences with a length between 19 and 30 bp and a Tm value between 67 and 72°C were rand...
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