US 2023/0416722 A1 · APPARATUS AND METHOD FOR PRETREATMENT OF MICROBIAL SAMPLES

Abstract

Methods and devices are provided for pretreatment of a sample containing microbial cells. In some embodiments, the pretreatment of the sample is performed via the initial selective lysis, within a sample pretreatment vessel, of non-microbial cells (such as blood cells) and the subsequent centrifugal separation of the sample to remove the resulting debris and concentrate the microbial cells. An immiscible and dense cushioning liquid may be included for collecting the microbial cells adjacent to the liquid interface formed by the cushioning liquid upon centrifugation of the pretreatment vessel. After removal of a substantial quantity of the supernatant, resuspension of the collected microbial cells, and re-establishment of the cushioning liquid interface, at least a portion of the remaining suspension may be removed without substantially removing the cushioning liquid. One or more intermediate wash cycles may be performed prior to extraction of the remaining suspension, which provides a “pretreated” sample.

Claims18

Dependent claims

1-4. (canceled)

An automated system for processing a positive blood culture sample, the automated system comprising:

a centrifugal processing receptacle comprising:
a main body having a proximal opening and a closed distal end, said main body comprising an inner chamber accessible though the proximal opening, wherein a surface defining said inner chamber comprises a first surface and a second surface, wherein said second surface is conical in shape and extends distalward relative to said first surface, and wherein a proximal portion of said second surface resides adjacent to a distal portion of said first surface;
and
a cap for closing the proximal opening during centrifugation of said centrifugal processing receptacle;
a rotor assembly configured to support said centrifugal processing receptacle;
a liquid handling subsystem capable of automated dispensing and aspiration;
control hardware operatively coupled to said rotor assembly and said liquid handling subsystem, said control hardware being configured to perform operations comprising:
controlling said liquid handling subsystem to combine the positive blood culture sample with a lysis reagent to form a mixture within said inner chamber of said centrifugal processing receptacle;
controlling said rotor assembly to centrifuge said centrifugal processing receptacle with the mixture residing within said inner chamber and the proximal opening closed by said cap, such that microbial cells are collected within a collection region residing between the proximal portion of said second surface and the distal portion of said first surface;
controlling said liquid handling subsystem to access said inner chamber and aspirate a first supernatant from said inner chamber;
controlling said liquid handling subsystem to dispense a wash buffer into said inner chamber;
controlling said rotor assembly to centrifuge said centrifugal processing receptacle with the wash buffer and the microbial cells residing within said inner chamber of said centrifugal processing receptacle and the proximal opening closed by said cap;
and
controlling said liquid handling subsystem to access said inner chamber and aspirate a second supernatant from said inner chamber, such that said inner chamber comprises purified and concentrated microbial cells.

11 more elements

A method of processing a positive blood culture sample, the method comprising:

providing a centrifugal processing receptacle comprising:
a main body having a proximal opening and a closed distal end, the main body comprising an inner chamber accessible though the proximal opening, wherein a surface defining the inner chamber comprises a first surface and a second surface, wherein the second surface is conical in shape and extends distalward relative to the first surface, and wherein a proximal portion of the second surface resides adjacent to a distal portion of the first surface;
and
a cap for closing the proximal opening during centrifugation of the centrifugal processing receptacle;
employing a liquid handling subsystem to form a mixture comprising the positive blood culture sample and a blood lysis reagent within the inner chamber;
employing a rotor assembly configured to support the centrifugal processing receptacle to centrifuge the centrifugal processing receptacle with the proximal opening closed by the cap, such that microbial cells are collected within a collection region residing between the proximal portion of the second surface and the distal portion of the first surface;
employing the liquid handling subsystem to access the inner chamber and aspirate a first supernatant from the inner chamber;
employing the liquid handling subsystem to dispense a wash buffer into the inner chamber;
employing the rotor assembly to centrifuge the centrifugal processing receptacle with the wash buffer and the microbial cells residing within the inner chamber of the centrifugal processing receptacle and the proximal opening closed by the cap;
and
employing the liquid handling subsystem to access the inner chamber and aspirate a second supernatant from the inner chamber, such that the inner chamber comprises purified and concentrated microbial cells.

8 more elements

An automated system for processing a liquid containing microbial cells, said automated system comprising:

a centrifugal processing receptacle comprising:
a main body having a proximal opening and a closed distal end, said main body comprising an inner chamber accessible though the proximal opening, wherein a surface defining said inner chamber comprises a first surface and a second surface, wherein said second surface is conical in shape and extends distalward relative to said first surface, and wherein a proximal portion of said second surface resides adjacent to a distal portion of said first surface;
and
a cap for closing the proximal opening during centrifugation of said centrifugal processing receptacle;
and
a rotor assembly configured to support said centrifugal processing receptacle;
and
control hardware operatively coupled to said rotor assembly, said control hardware being configured to perform operations comprising:
controlling said rotor assembly to centrifuge said centrifugal processing receptacle with the liquid residing within said inner chamber and the proximal opening closed by said cap, such that the microbial cells are collected within a collection region residing between the proximal portion of said second surface and the distal portion of said first surface.

6 more elements

A method of processing a blood sample containing microbial cells, the method comprising:

providing a centrifugal processing receptacle comprising:
a main body having a proximal opening and a closed distal end, the main body comprising an inner chamber accessible though the proximal opening, wherein a surface defining the inner chamber comprises a first surface and a second surface, wherein the second surface is conical in shape and extends distalward relative to the first surface, and wherein a proximal portion of the second surface resides adjacent to a distal portion of the first surface;
and
a cap for closing the proximal opening during centrifugation of the centrifugal processing receptacle;
forming a mixture comprising the blood sample and a blood lysis reagent within the inner chamber;
and
employing a rotor assembly configured to support the centrifugal processing receptacle to centrifuge the centrifugal processing receptacle with the proximal opening closed by the cap, such that the microbial cells are collected within a collection region residing between the proximal portion of the second surface and the distal portion of the first surface.

4 more elements

Dependencies are inferred from the claim text.

Facts

Application

18/208,713Filed 12 Jun 2023

Priority

30 Nov 2012

Published

28 Dec 2023

Term ends

Not estimatedThe source records don’t support an estimate.

Classification12 codes

C12N 15/1003
Processes for the isolation, preparation or purification of DNA or RNA: extracting or separating nucleic acids from biological samples, e.g. pure…
B01L 3/5021
With fluid transport, e.g. in multi-compartment structures: test tubes specially adapted for centrifugation purposes
B01L 3/5027
With fluid transport, e.g. in multi-compartment structures: by integrated microfluidic structures, i.e. dimensions of channels and chambers are…
C08F 220/286
And containing a polyether chain in the alcohol moiety: and containing polyethylene oxide in the alcohol moiety, e.g. methoxy polyethylene glycol…

Family

Application61/731,809Us provisional application · filed 30 Nov 2012

Application61/750,242Us provisional application · filed 8 Jan 2013

Application14/648,161Continuation in part · filed 28 May 2015

Application15/612,091Continuation · filed 2 Jun 2017

Application16/881,479Continuation · filed 22 May 2020

Application18/208,713Continuation

Application15/612,091Continuation in part

US 10,662,420 B2Related patent

US 11,702,647 B2Related patent

US 2016/0068897 A1Published application

US 2017/0275612 A1Published application

US 2020/0291386 A1Published application

US 9,707,555 B2Related patent

US 2023/0416722 A1This patent

From the US filing history. Foreign family members aren’t listed.

Apparatus and method for pretreatment of microbial samples

Life of this patent

Apparatus and method for pretreatment of microbial samples

Life of this patent

Abstract

Claims18