Primers, probe and kit for detecting mycoplasma genitalium and detection method
A technology of Mycoplasma genitalium and detection method, which is applied in biochemical equipment and methods, microorganism-based methods, microorganisms, etc., can solve the problems of low degree of automation, long detection time, and few kits, and achieves short detection time and high efficiency. Sensitive, highly specific effects
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Embodiment 1
[0038] Embodiment 1 Primer and probe of the present invention
[0039] The sequences of primers and probes of the present invention are as follows:
[0040] Table 1 Primer and probe sequences of the present invention
[0041]
Embodiment 2
[0042] Embodiment 2 Kit of the present invention
[0043] The components of the kit of this embodiment are shown in Table 2.
[0044] Table 2
[0045]
[0046] Wherein, reaction solution 1 comprises base buffer, cation, enhancer and enzyme, and described base buffer is the Tris-HCl buffer solution containing BSA, and described cation is Mn 2+ 、K + One or both of them, the enhancer includes one or more of glycerol, DMSO, Tween20, Tricine X-100, potassium acetate and betaine, and the enzyme is a bifunctional enzyme and / or UNG enzyme
Embodiment 3
[0047] Embodiment 3 utilizes the method for kit of the present invention to detect Mycoplasma genitalium
[0048] (1) Sample preparation: extract the nucleic acid of the sample to be tested, and prepare negative and positive quality controls at the same time,
[0049](2) PCR amplification system:
[0050] Table 3 PCR amplification system
[0051]
[0052] In the PCR amplification system, the concentration of Primer 1-Primer 4 is 20-35 pmol / μl, the concentration of Probe 1-Probe 2 is 10-20 pmol / μl, and the concentration of each component in the reaction solution 1 is: bifunctional enzyme 5- 10U / ul, UNG enzyme 0.1~0.5U / ul, 20mM dNTP, 2.8% glycerol, 3.7% DMSO, 0.01% Tween 20, 60mM Tricine X-100, 110mM potassium acetate, 0.5M betaine, basic buffer; PCR reaction Solution 2 contains 20~30mM MnCl 2 , P300.
[0053] (3) PCR amplification detection:
[0054] The amplification system in Table 2 was tested on the computer. The amplification program was 50°C for 2 minutes, 96°C fo...
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