45 results about "Resazurin" patented technology

The invention discloses a method of screening a 1-deoxyxylulose-5-phosphate racemase inhibitor from a marine microorganism fermentation broth. The method includes: 1) a step of preparing a marine microorganism fermentation broth sample, namely, a step of inoculating a fungus, fermenting, extracting metabolites, concentrating and eluting; 2) a step of designing a primer and performing PCR amplification by adopting the whole genome of vibrio vulnificus as a template, cloning a vector, converting escherichia coli, inducing expression of a target protein, and subjecting the target protein to elution, dialysis, ultrafiltration and desalting to obtain His-DXR; 3) a step of reacting a bacterial suspension, a resazurin solution and the marine microorganism fermentation broth sample obtained by the step 1), measuring the fluorescence value, calculating the inhibition rate through an equation, and screening the fraction the inhibition rate of which is higher than 80%; and 4) reacting the fraction screened in the step 3) and the His-DXR obtained in the step 2), calculating the inhibition rate, and screening the fraction the inhibition rate of which is not less than 50%. The method is simple and accurate, and lays a foundation for development of novel antibacterial medicines adopting Dxr as a therapeutic target or development of leading compounds.

The invention discloses a high-sensitivity and high-selectivity fluorescent analysis method by introducing morpholine oligonucleotide to detect a specific DNA fragment. The method is as below: using morpholine oligonucleotide covalently attached to the surface of a magnetic microspheres as a capture probe, hybridizing the morpholine oligonucleotide with a long target DNA fragment; hybridizing an un-hybridized segment of the target DNA signal with a signal DNA with terminal modified with biotin; using thecombination effect of streptavidin and biotin, introducing alkaline phosphatase into the terminal of a signal probe DNA; catalyzing L-ascorbic acid 2-phosphate to generate L-ascorbic acid by alkaline phosphatase; and reducing resazurin by L-ascorbic acid to form strong fluorescent resorufin. The fluorescence intensity is positively correlated with the target DNA; through fluorescence analysis, high-sensitivity and high-selectivity detection of DNA fragments is achieved; and the method can be used in the detection of DNA samples in the fields of molecular recognition, clinical diagnosis, environmental monitoring and food safety.

The invention discloses a mycobacterium tuberculosis medicament sensitive phenotype detection method, which comprises the following steps of: 1) adding 2.5 to 6mul of 0.2 percent of resazurin dissolved by using methanol with mass percentage concentration into the inner side of a tube cover of a sterile centrifuge tube, and sterilizing the centrifuge tube after the methanol is volatilized; 2) inoculating 5 to 50mg of mycobacterium tuberculosis to be detected into a 7H9-S culture medium, regulating the concentration of the bacteria solution by adopting turbidimetry, regulating the concentration of the bacteria solution to the concentration of a Mcburney turbidimetric 1 tube by using sterile water or 0.9-percent physiological saline, and diluting the bacteria solution by 20 times by using the 7H9-S culture medium; 3) inoculating 50 to 150mu of diluted bacteria solution into the centrifuge tube of the step 1); 4) adding 100mul of anti-tubercular medicament solution diluted by 7H9-S and to be detected into the centrifuge tube of the step 3), wherein the concentration of the medicament to be detected is 0.01mug / ml to 2.5mg / ml; 5) after the centrifuge tube of the step 4) is covered closely, culturing the solution for 6 to 7 days at the temperature of 37 DEG C; and 6) inverting and oscillating the centrifuge tube of the step 5), and culturing the solution for 12 to 48 hours at the temperature of 37 DEG C after the solution becomes blue. Proved by experiments, in the mycobacterium tuberculosis medicament sensitive phenotype detection method and application of the mycobacterium tuberculosis medicament sensitive phenotype detection method in anti-tubercular medicament screening, the method has low workload and short detection time; compared with a traditional L-J medium medicament sensitive detection method, the accuracy reaches 90 to 99 percent; and the method has good biological safety, does not cause laboratory propagation of tuberculosis, is suitable for large-scale.

The invention discloses an Ackerman myxobacteria culture medium and a preparation method thereof. The Ackerman myxobacteria culture medium per liter is prepared from the following raw materials: 0.4g of monopotassium phosphate, 0.53g of disodium hydrogen phosphate, 0.3g of ammonium chloride, 0.3g of sodium chloride, 0.11g of calcium chloride, 0.1g of magnesium chloride hexahydrate, 4g of sodium bicarbonate, 0.5mg of resazurin, 1mL of an acid trace solution, 1mL of an alkali trace solution, 1mL of a vitamin solution, 37g of a brain heart infusion, 1.5g of xylooligosaccharide, 5g of cyclodextrin and 30g of solarbio protease-enzymolyzed egg white. The culture medium is capable of solving the problem of slow growth of Ackerman myxobacteria, and significantly improving the growth rate and the bacteria solution concentration of Ackerman myxobacteria.