Mycoplasma pneumoniae rapid detection kit and use method thereof

A technology of mycoplasma pneumoniae and detection kits, which is applied in the direction of microorganism-based methods, biochemical equipment and methods, and microorganism measurement/inspection. It can solve the problems that traditional detection methods cannot meet clinical diagnosis, and achieve intuitive observation and specificity. Strong, highly sensitive effects

Inactive Publication Date: 2010-03-10
ZHUHAI ENCODE MEDICAL ENG
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Problems solved by technology

[0005] The purpose of the present invention is to provide a method for the preparation and use of a Mycoplasma pneumoniae rapid detection kit (LAMP), to improve the diagnostic efficiency of Mycoplasma pneumoniae, save time, and solve the problem that traditional detection methods cannot meet the development requirements of clinical diagnosis. After several experiments and tests, a method for clinical Mycoplasma pneumoniae detection using loop-mediated constant temperature amplification technology was finally explored. This method has the characteristics of rapid detection, convenience, and low cost.

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Embodiment Construction

[0030] The following implementation examples do not illustrate the present invention, but should not be regarded as limiting the present invention.

[0031] Make Mycoplasma pneumoniae rapid detection kit (LAMP) according to the following formula:

[0032] (1) LAMP reaction tube

[0033] Contains 2.5 μL 10× ThermoPol reaction buffer, 1 μL 10 μmol / L primer 1, 1 μL 10 μmol / L primer 2, 1 μL 40 μmol / L primer 3, 1 μL 40 μmol / L primer 4, 3.5 μL 10 mmol / L dNTP, 1 μL 100 mmol / L magnesium sulfate , 5 μL 5mol / L betaine, 1 μL 12.5mmol / L manganese chloride, 1 μL 625 μmol / L calcein, 4 μL sterile double distilled water. Wherein, the 10×ThermoPol reaction buffer contains tris-hydrochloric acid of 200mM pH8.8, the potassium chloride of 100mM, the ammonium sulfate of 100mM, the magnesium sulfate of 20mM and the triton X-100 of 1%;

[0034] Wherein the mass ratio of dATP, dTTP, dCTP and dGTP contained in the dNTP is 1:1:1:1;

[0035] Primer 1 described therein: 5-ACCAATGCCATCAACCCG-3,

[003...

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Abstract

The invention relates to a mycoplasma pneumoniae rapid detection kit and a use method thereof, wherein the kit holds a loop-mediated isothermal amplification reaction tube and BstDNA polymerase; the reaction tube holds reaction buffer, dNTP, magnesium sulfate, a primer 1:5-ACCAATGCCATCAACCCG-3, a primer 2:5-TACCGGCGTAACGCAAAG-3, a primer 3:5-ATTTTCACCCGTGAGGGGGAGTTTTCGCTTAACCCCGTGAACG-3, a primer4:5-ACAGCGCTAAGGGCATCACTGTTTTTCAAAGCCGCTTCGGTTC-3, lycine, manganese chloride and calcein. The method for detecting mycoplasma pneumoniae comprises the following steps: extraction of a sample to be detected or bacterium DNA to be detected, loop-mediated isothermal amplification reaction of mycoplasma pneumoniae and colour development detection. The invention has the characteristics of accurate detection, high sensitivity, strong specificity, simpleness, convenience and rapidness.

Description

technical field [0001] The invention relates to a method for rapid detection of bacterial samples using a loop-mediated constant temperature amplification (LAMP) technology, in particular to a rapid detection kit (LAMP) for mycoplasma pneumoniae and a use method thereof. Background technique [0002] Mycoplasma pneumoniae (MP) is the pathogen of human mycoplasma pneumonia and one of the main pathogens causing respiratory tract infection and atypical pneumonia in children. More than 1 / 3 of non-bacterial pneumonia is caused by MP. MP is also an important pathogen of community-acquired pneumonia, and the infection rate in densely populated places is as high as 50%. Rapid and accurate detection of Mycoplasma pneumoniae is a prerequisite for effective prevention of Mycoplasma pneumoniae infection. At present, the clinical identification of Mycoplasma pneumoniae is usually carried out using traditional physiological and biochemical methods. However, due to the high requirements o...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68G01N21/78C12R1/35
Inventor 孙宜峰曾冰冰胡荣刘义江
Owner ZHUHAI ENCODE MEDICAL ENG
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