公开(公告)号：US20200149085A1

公开(公告)日：2020-05-14

申请号：US16734826

申请日：2020-01-06

Applicant: BECTON DICKINSON AND COMPANY

Inventor： Liping Feng ,  William B. Brasso ,  Susan M. Kircher ,  Vanda White ,  Song Shi ,  Xiao Mo ,  Tuan-Linh Ngoc Nguyen ,  Adrien P. Malick ,  Jon E. Salomon ,  John D. Mantlo

IPC: C12Q1/04 ,  G01N33/68 ,  G01N33/483

Abstract: 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.

公开(公告)号：US10557162B2

公开(公告)日：2020-02-11

申请号：US15472759

申请日：2017-03-29

Applicant: Becton, Dickinson and Company

Inventor： Liping Feng ,  William B. Brasso ,  Susan M. Kircher ,  Vanda White ,  Song Shi ,  Xiao Mo ,  Tuan-Linh Ngoc Nguyen ,  Adrien P. Malick ,  Jon E. Salomon ,  John D. Mantlo

IPC: C12Q1/04 ,  G01N33/68 ,  G01N33/483 ,  H01J49/16 ,  H01J49/40

Abstract: 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.

公开(公告)号：US10519482B2

公开(公告)日：2019-12-31

申请号：US14381388

申请日：2013-02-28

Applicant: Becton, Dickinson and Company

Inventor： Susan M. Kircher ,  Vanda White ,  William B. Brasso ,  Dyan Luper ,  James Y. Zhou ,  Julie L. Rosales ,  Jeffery H. Bruton ,  John D. Mantlo ,  Adrien P. Malick ,  Donald R. Callihan ,  Ben Turng ,  Liping Feng ,  Curtis M. Gosnell ,  Patrick Shawn Beaty ,  John P. Douglass

IPC: C12Q1/14 ,  C12Q1/24 ,  C12N1/02 ,  C12N1/20 ,  C12N1/04 ,  C12Q1/06

Abstract: Various embodiments disclosed herein provide for reagents and methods for rapidly isolating viable microbial cells, including S. pneumoniae, 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.

公开(公告)号：US20150125895A1

公开(公告)日：2015-05-07

申请号：US14381388

申请日：2013-02-28

Applicant: Becton, Dickinson and Company

Inventor： Susan M. Kircher ,  Vanda White ,  William B. Brasso ,  Dyan Luper ,  James Y. Zhou ,  Julie L. Rosales ,  Jeffery H. Bruton ,  John D. Mantlo ,  Adrien P. Malick ,  Donald R. Callihan ,  Ben Turng ,  Liping Feng ,  Curtis M. Gosnell ,  Patrick Shawn Beaty ,  John P. Douglass

IPC: C12Q1/14 ,  C12N1/20 ,  C12Q1/24

CPC classification number: C12Q1/14 ,  C12N1/02 ,  C12N1/20 ,  C12Q1/24

Abstract: Various embodiments disclosed herein provide for reagents and methods for rapidly isolating viable microbial cells, including S. pneumoniae, 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.

Abstract translation: 本文公开的各种实施方案提供了用于从阳性血培养样品中快速分离存活的微生物细胞（包括肺炎链球菌）的试剂和方法。 所得到的微生物颗粒可用于鉴定和基于生长的方法如抗微生物敏感性测试。 本文所述的缓冲剂可以含有碱溶液，非离子洗涤剂，硫醇和任选的氯化铵。 所公开的方法提供了一种用于仅从一个样品制备管和离心同时从PBC样品中快速分离和浓缩活的微生物的方法，同时从哺乳动物血细胞中除去可能干扰鉴定方法的细胞碎片。

公开(公告)号：US08603769B2

公开(公告)日：2013-12-10

申请号：US13647072

申请日：2012-10-08

Applicant: Becton, Dickinson and Company

Inventor： Liping Feng ,  William B. Brasso ,  Song Shi ,  Ben Turng ,  Susan M. Kircher ,  Vanda White ,  Dyan Luper ,  Julie Rosales ,  Gretta Campbell ,  Adrien Malick

IPC: C12Q1/04 ,  C12Q1/24

CPC classification number: C12Q1/24

Abstract: 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 S. pneumoniae.

Abstract translation: 本发明的方法包括从用于下游分析如鉴定和抗微生物敏感性测试（“AST”）的阳性血培养（“PBC”）样品中分离完整的活的微生物。 所述方法包括收集一部分PBC样品，加入含胆碱的溶液，裂解血细胞，分离活微生物，以及进行分离的活的微生物的下游分析。 该方法可以应用于各种革兰氏阳性细菌，革兰氏阴性细菌和/或酵母，特别是肺炎链球菌菌株。