Method for enriching filamentous fungi from blood sample

Abstract

The invention relates to a method for enriching filamentous fungi from a blood sample. The method comprises a step (1) of extracting 1-2 ml of blood sample from a blood culture positive culture bottle, and injecting the blood sample into a centrifuge tube; a step (2) of adding 50-150 mg of quartz sand and 10-20 ml of enriched diluent into the centrifuge tube, performing mixing evenly, standing, and performing centrifugation to obtain sediment; a step (3) of vibrating the sediment, performing centrifugation, and adding sodium dodecyl sulfate SDS solution and sterilized distilled water, and performing vibration to remove the quartz sand; performing centrifugation and washing, and obtaining the filamentous fungi. The method is high in efficiency of enriching the filamentous fungi, impurities in the enriched fungi are fewer, and the method can be used for fungus nucleic acid extraction and can also be used for direct identification of the fungi through a mass spectrum technology and have wide application space.

Description

technical field [0001] The invention belongs to the field of blood sample processing, in particular to a method for enriching filamentous fungi from blood samples. Background technique [0002] With the acceleration of the aging process of the social population, the number of people suffering from senile chronic diseases is increasing. At the same time, broad-spectrum antibiotics are widely used, and invasive medical operations such as catheter interventional therapy and tissue biopsy are increasing, as well as tumors, AIDS, leukemia, and organ transplants. The number of immunocompromised patients continues to expand. Therefore, in recent years, the incidence of deep fungal infection has increased significantly, especially the incidence of deep filamentous fungal infection is increasing year by year, and its incidence is even higher than that of yeast infection in some specific populations. Due to the lack of characteristic clinical symptoms in the early stage of deep funga...

AI Technical Summary

Problems solved by technology

Filamentous fungi, however, form thin hyphae in the bloodstream and are present in small amounts in the patient's circulating blood and grow slowly

Mycelium is different from bacteria or fungal spores. Using the same experimental conditions, it is difficult to efficiently enrich and isolate such mycelia from blood.

[0005] At present, in the commonly used clinical blood sample digestion treatment solution, the hemolytic agent is mostly ammonium chloride, ammonium acetate, etc., which are toxic or slightly toxic substances, such as irritating the skin, mucous membranes, eyes, nasal cavity, and throat, and damaging the eyes; high concentrations stimulate the lungs, which can cause Pneumonia; moreover, there is a high concentration of protein in the blood sample, and even lymphocytes continue to grow and multiply in the blood culture bottle. Ammonium chloride and ammonium acetate are likely to cause protein precipitation, making the solution turbid and increasing the viscosity. Facilitate the isolation of filamentous fungi

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