26-09-2019 дата публикации
Номер: US20190293646A1
The invention provides methods for rapid isolation of microorganisms from positive culture sterile body fluids, including blood, cerebrospinal fluid (CSF), pleural fluid, ascitic fluid, pericardial effusion, joint cavity fluid, vitreous fluid, and amniotic fluid for mass spectrometry identification. Whenever the subject of blood culture is discussed, the intended sample used is always related to blood sample. However, it is also necessary to be aware that other than the blood sample, sterile fluids can also be inoculated as samples for blood culture testing. Among the sterile fluids that are commonly known are CSF, pleural fluid, ascitic fluid, pericardial effusion, joint cavity fluid, vitreous fluid, amniotic fluid etc. The methods involve combining micro-volume positive blood culture sample with detergent solution to lyse human blood cells, then isolating the microorganism by differential centrifugation process that first removes interfering substances such as charcoal (when present), resins and human blood cellular debris through a low speed centrifugation, then isolates the microorganisms in the sample supernatant through a fast centrifugation. The methods not only apply to regular blood culture media but also apply to antimicrobial removal containing media such as resin containing BD BACTEC™ Plus-Aerobic media and charcoal-containing Biomerieux BacT/Alert® FA media. In addition, the methods can isolate a variety of Gram-positive bacteria, Gram-negative bacteria, and yeast in clinical settings. The isolated microorganism(s) from positive blood culture can be used for multiple downstream analyses, including identification of the microorganism(s) by mass spectrometry, phonotypical, or molecular identification methods.
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