Method of identifying cyanobacteria cells by means of time of flight mass spectrometry
A technology of time-of-flight mass spectrometry and cyanobacteria, which is applied in the field of proteomics and environmental analytical chemistry, can solve the problems that there is no suitable method for cyanobacteria identification, and achieve the effects of small sample dosage requirements, simplified steps, and strong characteristic peak signals
Pending Publication Date: 2017-06-13
SOUTHEAST UNIV
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Problems solved by technology
Matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS) is a soft ionization technique, and this method of analytical chemistry has been successfully applied in the rapid and accurate identification of other microorganisms, but the identification of cyanobacteria has not yet been reported. Appropriate methods, especially pretreatment steps, matrix selection and protein crystallization methods have always been the focus of research
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Embodiment 1
[0029] In this example, water samples were taken outside the Meiyuan Water Plant in Inner Taihu Lake on May 30, 2016, during the outbreak of water blooms, as samples to be tested, and 100 mL of water samples were collected in plastic bottles through the phytoplankton net. Transfer the water samples into 15ml centrifuge tubes and number them ① and ② to prepare for the subsequent comparison and reference. After the marked samples are debubbled slightly by ultrasonic waves, put them in a centrifuge at 9100g and centrifuge at 4°C for 10 minutes. After centrifugation, remove the supernatant, use a pipette gun to remove the bottom sediment into a 1.5mL centrifuge tube (weigh the self-weight number of the centrifuge tube in advance and record it), and centrifuge at 20400g, 4°C for 10 minutes. After the previous step, add 160μL of Mix TMA-1 buffer, TMA-1 buffer contains 10mM trishydrochloride (pH 7.8), 30mM ammonium chloride, 10mM magnesium chloride and 6mM 2-mercaptoethanol, prepared ...
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The invention discloses a method of identifying cyanobacteria cells by means of time of flight mass spectrometry. The method comprises the following steps of: (1) pre-treating a to-be-identified cyanobacteria cell sample; (2) dropping the sample treated in the step (1) onto a sample board, then dropping a substrate solution, and airing indoors; and (3) putting the sample plate obtained in the step (2) into a time of flight mass spectrometry spectrometer for detection, and comparing the detected map with the selected characteristic ribosomal protein theoretical peak value by taking a ribosomal protein as a bio-identifier to identify the types of the cyanobacteria cells. By applying the method, the cyanobacteria cells can be quickly, simply and directly detected and sorted and identified, the result is accurate, the manpower and time cost are reduced, the testing program can be standardized, and finally, an online programmed operation of identifying blue-green algae is realized.
Description
technical field [0001] The invention relates to a detection technology for directly measuring cyanobacteria cells by time-of-flight mass spectrometry, and belongs to the technical fields of proteomics and environmental analysis chemistry. Background technique [0002] Cyanobacteria, as primitive and ancient algae prokaryotes, tend to multiply in large numbers in eutrophic lakes, reservoirs and rivers to form cyanobacteria blooms when the temperature is high, especially in summer, under suitable hydrological and meteorological conditions. Many cyanobacteria, especially the most common Microcystis aeruginosa in freshwater cyanobacteria, produce toxic secondary metabolites represented by microcystins, which cause great damage to the liver and skin of humans and animals, and seriously endanger the water environment and human health and well-being. life safety. Therefore, it is particularly important to identify the dominant species of Microcystis aeruginosa in cyanobacterial bl...
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IPC IPC(8): G01N27/62
CPCG01N33/6848
Inventor 孙丽伟江文静
Owner SOUTHEAST UNIV