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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

Inactive Publication Date: 2020-03-20
AUTOBIO DIAGNOSTICS CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, there are few kits for detecting MG based on PCR technology in China, and the degree of automation is not high, the detection time is long, and the false negative is high. Therefore, it is necessary to develop a simple and fast reagent with high sensitivity, short detection time, low false positive The box is the problem that needs to be solved

Method used

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  • Primers, probe and kit for detecting mycoplasma genitalium and detection method
  • Primers, probe and kit for detecting mycoplasma genitalium and detection method
  • Primers, probe and kit for detecting mycoplasma genitalium and detection method

Examples

Experimental program
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Effect test

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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Abstract

The invention relates to the technical field of medical detection, in particular to primers for detecting mycoplasma genitalium, a probe for detecting mycoplasma genitalium, a kit for detecting mycoplasma genitalium and a detection method. The primers and the probe comprise a primer pair of nucleotide sequences shown in SEQ ID NO:1-2 as shown in the description and a probe of a nucleotide sequenceshown in SEQ ID NO:3 as shown in the description. The provided primers and probe can specifically bound to a mycoplasma genitalium outer membrane protein B gene and a human beta-globin gene, the detection sensitivity and specificity are significantly improved, the positive rate of low-value samples reaches 100%, and detection results are accurate and reliable so that the primers, probe and kit for detecting mycoplasma genitalium and the detection method can be applied to clinical detection of mycoplasma genitalium.

Description

technical field [0001] The invention relates to the technical field of medical detection, in particular to a primer, a probe, a kit and a detection method for detecting mycoplasma genitalium. Background technique [0002] Mycoplasma genitalium (Mycoplasma genitalium, MG) belongs to Mycoplasma family Mollicola, is one of the pathogens that cause non-gonococcal urethritis (NGU). In addition, MG is closely related to sexually transmitted diseases such as pelvic inflammatory disease (PID), obstetric complications, prostatitis, epididymitis, and balanitis. In recent years, studies have found that MG infection may also be related to the occurrence and development of tumors and human immunodeficiency virus (HIV). [0003] The 2016 European guidelines for Mycoplasma genitalium infection state that the prevalence of Mycoplasma genitalium in men with non-Chlamydial nongonococcal urethritis (NCNGU) ranges from 10% to 35%. In China, the infection rate of Mycoplasma genitalium in STD c...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/689C12Q1/6851C12N15/11C12Q1/04C12R1/35
CPCC12Q1/689C12Q1/6851C12Q2600/166C12Q2531/113C12Q2545/101C12Q2563/107
Inventor 胡进李振红李静静杜美鲁清月付光宇吴学炜苗拥军
Owner AUTOBIO DIAGNOSTICS CO LTD
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