Becton Dickinson and Company Limited

Vanda White

Adrien P Malick

Susan M Kircher

William B Brasso

Liping Feng

John D Mantlo

Song Shi

Ben Turng

Julie L Rosales

Jon E Salomon

Xiao Mo

Dyan Luper

Tuan-Linh Ngoc Nguyen

Patrick Shawn Beaty

Donald Ray Callihan

Curtis M Gosnell

James Y Zhou

Jeffery H Bruton

John P Douglass

Gretta Campbell

Donald R Callihan

Shawn Beaty

Various embodiments disclosed herein provide for reagents and methods for rapidly isolating viable microbial cells, including, from positive blood culture samples. The resulting microbial pellet can be used for both identification and growth-based methods such as antimicrobial susceptibility testing. The buffers described herein may contain a base solution, non-ionic detergents, thiols, and optionally, ammonium chloride. The disclosed methods provide a process for rapidly isolating and concentrating viable microorganism(s) from PBC samples using only one sample preparation tube and centrifugation while removing cellular debris from the mammalian blood cells that may interfere with identification methods.

Formulations and process for isolating viable microorganisms from positive blood cultures

Rapid methods that identify sepsis-causing bacteria or yeast aid the physician in critical therapeutic decision-making, thus decreasing patient mortality rates. The methods described herein employ plating microorganisms directly on to a MALDI-MS plate, adding concentrated formic acid, and identifying the microorganism by mass spectrometry. Optionally, an organic solvent may be combined with the formic acid, or added to the sample before or after the concentrated formic acid is added thereto. The methods enable direct extraction of proteins from microorganisms without the need for liquid protein extraction methods and yields positive identification results for gram-positive bacteria, gram-negative bacteria and yeast in minutes.

Identification of microorganisms using MALDI-TOF-MS on-plate extraction

Various embodiments disclosed herein provide for reagents and methods for rapidly isolating viable microbial cells, including, from positive blood culture samples. The resulting microbial pellet can be used for both identification and growth-based methods such as antimicrobial susceptibility testing. The buffers described herein may contain a base solution, non-ionic detergents, thiols, and optionally, ammonium chloride. The disclosed methods provide a process for rapidly isolating and concentrating viable microorganism (s) from PBC samples using only one sample preparation tube and centrifugation while removing cellular debris from the mammalian blood cells that may interfere with identification methods.

Methods of the invention include the isolation of intact, viable microorganism(s) from positive blood culture ('PBC') samples for use in downstream analyses such as identification and antimicrobial susceptibility testing ('AST'). The methods involve collecting a portion of the PBC sample, adding a choline-containing solution, lysing the blood cells, isolating the viable microorganism, and performing downstream analysis of the isolated, viable microorganism. The methods can be applied to a variety of gram-positive bacteria, gram-negative bacteria, and/or yeast, and particularly to strains of

Method for direct and rapid identification of microorganisms and antimicrobial susceptibility testing from positive blood cultures

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Baltimore, MD, United States of America

Vanda White