Mycoplasma urealytium virus nucleic acid detection kit

A technology for detection of Ureaplasma urealyticum and viruses, which is applied in the determination/inspection of microorganisms, microorganisms, biochemical equipment and methods, etc. It can solve the problems of long detection time, poor specificity, and low detection sensitivity, and achieve improved sensitivity and specificity , the effect of short detection time

Inactive Publication Date: 2017-08-18
GUANGZHOU HEAS BIOTECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] The technical problem mainly solved by the present invention is to provide a ureaplasma virus nucleic acid detection kit, which solves the problems of low detection sensitivity, poor specificity and long detection time of common ureaplasma virus detection kits on the market

Method used

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  • Mycoplasma urealytium virus nucleic acid detection kit
  • Mycoplasma urealytium virus nucleic acid detection kit
  • Mycoplasma urealytium virus nucleic acid detection kit

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0035] Example 1 The operating method of the Ureaplasma urealyticum virus nucleic acid detection kit:

[0036] 1) DNA extraction:

[0037] Take 500~1000μl secretions into a 1.5ml centrifuge tube, centrifuge at 12000rpm for 1 minute, pour off the supernatant, then add 50μl of DNA extraction solution, boil at 100°C for 5 minutes, centrifuge the centrifuge tube at 12000rpm for 2 minutes, and take 10μl The supernatant was added to the PCR reaction tube;

[0038] 2) Rapid PCR:

[0039] In 50 μl fluorescent PCR system, 3U fast Taq enzyme, 12mM Tris-HCL, 45mM KCL, 0.5U UNG enzyme, 0.25mM dNTPS (U), 2.5mM MgCl2, 10pM each for SEQ1 and SEQ2, 3pM for SEQ3. 50°C for 3 minutes, 95°C for 5 minutes, 95°C for 5 seconds, 58°C for 15 seconds, the number of cycles is 40 cycles;

[0040] 3) Monitor the fluorescence change of the reaction system. If the fluorescence signal increases, there is Ureaplasma urealyticum virus infection.

Embodiment 2

[0041] Embodiment 2 Comparative experiments of the fast Taq enzyme system used in the present invention and the common Taq enzyme system:

[0042] 1) DNA extraction:

[0043] Take 500~1000μl secretions into a 1.5ml centrifuge tube, centrifuge at 12000rpm for 1 minute, pour off the supernatant, then add 50μl of DNA extraction solution, boil at 100°C for 5 minutes, centrifuge the centrifuge tube at 12000rpm for 2 minutes, and take 10μl The supernatant was added to the PCR reaction tube;

[0044] 2) Rapid Taq Enzyme System PCR:

[0045] In 50 μl fluorescent PCR system, 3U fast Taq enzyme, 12mM Tris-HCL, 45mM KCL, 0.5U UNG enzyme, 0.25mM dNTPS (U), 2.5mM MgCl2, 10pM each for SEQ1 and SEQ2, 3pM for SEQ3. 50°C for 3 minutes, 95°C for 5 minutes, 95°C for 5 seconds, 55°C for 15 seconds, the cycle number is 40 cycles;

[0046] 3) Common Taq enzyme system PCR:

[0047] In 50 μl fluorescent PCR system, 3U hot start Taq enzyme, 12mM Tris-HCL, 45mM KCL, 0.5U UNG enzyme, 0.25mM dNTPS (U...

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Abstract

The invention discloses a mycoplasma urealytium virus nucleic acid detection kit which comprises DNA extract, a quick Taq enzyme system and a mycoplasma urealytium virus detection primer probe. The mycoplasma urealytium virus detection primer probe is composed of a specific primer designed for mycoplasma urealytium and a corresponding fluorescent probe, and one end of the probe is marked with MGB group. The kit is used for qualitatively detecting whether a sample is infected with mycoplasma urealytium virus and is simple in detection operation. By using the MGB probe, sensitivity and specificity of detection are improved effectively. By using the quick Taq enzyme system, time needed for reaction is shortened remarkably, and working efficiency is improved.

Description

technical field [0001] The invention relates to a pathogen nucleic acid detection kit, in particular to a Ureaplasma urealyticum virus nucleic acid detection kit. Background technique [0002] Ureaplasma urealyticum (UU) is one of the common parasites in the human genitourinary tract, which can cause disease under certain circumstances. It is mainly transmitted through sexual life, and can also be transmitted vertically through the placenta or spread upward from the lower genital tract infection of pregnant women, causing intrauterine infection. Its infection can lead to a series of adverse consequences such as infertility, miscarriage, premature delivery, and fetal growth retardation. . The number of UU colonization in the human body has two peak periods, that is, the mother's birth canal infects the newborn during childbirth, and then decreases rapidly, and gradually increases after sexual life. The size of UU is generally 50-300 nnl , the structure is simple, there is ...

Claims

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Application Information

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IPC IPC(8): C12Q1/70C12Q1/68C12R1/93
CPCC12Q1/701
Inventor 刘淑园陈华云丁渭肖湘文陆同山
Owner GUANGZHOU HEAS BIOTECH CO LTD
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