Application of miR-331-5p as acute HIV infection detection marker
An acute and material technology, applied in the field of acute HIV infection detection markers, can solve problems such as high price, and achieve the effects of convenient operation, high sensitivity and favorable clinical application.
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Embodiment 1
[0040] Example 1 Practical verification of the correlation between miR-331-5p expression in clinical plasma samples and acute HIV infection
[0041] 1. Material and Specimen Collection Description
[0042] The samples were all from the dynamic follow-up cohort of high-risk gay men in You'an Hospital, followed up once every 2 months. After the whole blood was collected, the plasma and peripheral blood mononuclear cells were separated and frozen. The samples of 20 cases of acute HIV infection before and after infection were selected. , These infected persons were excluded from the merger of syphilis, hepatitis B and C.
[0043] 2. Plasma miRNA extraction
[0044] Plasma RNA was extracted and purified by Qiagen's miRNeasySerum / PlasmaKit kit according to the kit instructions. The specific steps are as follows:
[0045] 2.1 Thaw frozen plasma samples.
[0046] 2.2 Add 500 μL of QIAzolLysis Reagent to each 100 μL sample, and use a pipette to repeatedly blow and mix.
[0047] 2.3...
Embodiment 2
[0082] Embodiment 2 Preparation and use method of kit of the present invention
[0083] The composition of kit of the present invention:
[0084] 1. The primer sequence used to detect the expression level of miR-331-5p, the detection primer sequence is shown in 1 μL of the upstream primer SEQIDNo.3 (10 μM) and 1 μL of the downstream primer SEQIDNo.4 (10 μM).
[0085] 2. The PCR reaction solution can be a composition known to those skilled in the art. The recommended composition of the present invention is: 12.5 μL of PCR reaction solution, which contains SybergreenI fluorescent dye 0.5-1X, DNA polymerase 100U / mL, dNTPs0.2mM, trimethylol Aminomethane hydrochloride 16.5mM, potassium chloride 89.3mM, magnesium chloride 6mM and sterile water 5.5μL.
[0086] The method for using the kit of the present invention:
[0087] 1. Extraction of miRNA in the sample to be tested: Purchase Qiagen's miRNeasySerum / PlasmaKit kit and follow the kit instructions. Refer to the plasma miRNA extr...
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