IP Library Granted Patent US 8,877,458
Granted Patent B2
US 8,877,458 · App. 12/525,467 · Granted Nov 4, 2014

Method of detecting bacterial contamination using dynamic light scattering

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Quick Facts
Patent No.
US 8,877,458
App. No.
12/525,467
Granted
Nov 4, 2014
Kind
B2
Abstract

Methods of detecting bacterial contamination in a platelet concentrate are performed using a dynamic light scattering (DLS) instrument and a sample holder. A sample of platelet concentrate can be held vertically or horizontally in a capillary in the sample holder. Alternatively, novel platelet storage bags modified to include an optically translucent window can be held within another variant of the sample holder. Still alternatively, platelet storage bags having one or more tubes detachably appended to the bag can be used. A sample is drawn off into an appended tube for placement directly into the sample holder. This method provides a number of related, non-invasive techniques for detecting whether bacteria has contaminated a platelet concentrate. Contamination indicators include a population of particles different from platelets, microparticles or proteins, bad-quality platelets, i.e. low DLS score, and very high or very low scattering intensity.

Claims (31)

1. A method for detecting bacteria in a sample, the method comprising:

a) placing the sample in a dynamic light scattering (DLS) instrument;

b) collecting DLS measurements from the sample by determining a relative intensity of scattered light and establishing a particle size distribution based on the DLS measurements, wherein the particle size distribution comprises at least distinct first and second populations of particles corresponding to first and second particles in the sample having different particle sizes, wherein said first population corresponds to platelets;

c) comparing the first population to the second population and based upon said comparison,

making determination that a quantity of bacteria is present in the sample when the second population exceeds a predetermined threshold; and

d) if a determination has been made according to step c) that a quantity of bacteria is present in the sample, determining the quantity of bacteria present in the sample by comparison of the second population to the first population.

2. The method as claimed in claim 1 wherein each of the at least first and second distinct populations of particles in the particle size distribution is represented by a discrete peak, each peak corresponding to a different type of particle in the sample; and

analyzing the discrete peaks

and estimating a quantity of bacteria in the sample by analyzing the relative intensity of the scattered light for a discrete peak in the particle size distribution corresponding to the bacteria.

3. The method as claimed in claim 2 wherein the step of analyzing the discrete peaks further comprises identifying platelets, microparticles, proteins and bacteria by comparing mean particle sizes to expected particle size ranges for platelets, microparticles, proteins and bacteria, respectively, based on previously obtained empirical data.

4. The method as claimed in claim 1 further comprising a step of identifying a specific species of bacteria in the sample.

5. The method as claimed in claim 1 further comprising: controlling a temperature of said sample.

6. The method as claimed in claim 5 wherein said controlling said temperature of said sample comprises:

incubating the sample at a temperature of either 37 degrees Celsius±2 degrees Celsius or 20 degrees Celsius±2 degrees Celsius.

7. The method as claimed in claim 5 wherein said controlling said temperature of said sample comprises:

temperature cycling the sample between 37 and 20 degrees Celsius.

8. A method of detecting bacterial contamination in a platelet sample containing microparticles, the method comprising steps of:

obtaining DLS measurements from the platelet sample, said DLS measurements being obtained while the temperature of said platelet sample is between 37 and 20 degrees Celsius±2 degrees Celsius;

calculating a DLS score from the DLS measurements; determining whether the DLS score is below a predetermined threshold score; and

identifying the platelet sample as being bacterially contaminated when the DLS score is below the predetermined threshold score.

9. A method of detecting bacterial contamination in a platelet sample, the method comprising steps of:

obtaining DLS measurements from the platelet sample; said DLS measurements comprising a total scattering intensity derived from scattering intensities from plural distinct populations of particles in the sample, wherein each of said plural distinct populations is associated with an intensity of scattered light and wherein one of said plural distinct populations correspond to platelets in the sample and the other plural distinct populations correspond to other microparticles in the sample;

determining whether an intensity of scattered light from one of said other plural distinct populations is below a first predetermined intensity threshold;

determining whether the total scattering intensity is above a second predetermined intensity threshold and

identifying the platelet sample as being bacterially contaminated when the intensity of scattered light from said one of said other plural distinct populations is below the first predetermined intensity threshold or when the total scattering intensity is above the second predetermined intensity threshold.

10. A method of detecting bacterial contamination in a sample of platelets, the method comprising steps of:

obtaining DLS measurements on the sample of platelets by illuminating the sample with incident light and by collecting the scattered light;

establishing a single particle size distribution based on the scattered light, said single particle size distribution comprising plural data sets from the DLS measurements and wherein each data set corresponds to a cluster of particles in the sample that is distinct from other particles in the sample;

identifying a first data set from the plural data sets that corresponds to a cluster of particles known to correspond to platelets; identifying a second data set from the plural data sets that correspond to a cluster of particles on the particle size distribution that is distinct from the first data set;

comparing the first data set to the second data set and if the second data set exceeds a predetermined threshold value, making a determination that the second data set corresponds to bacteria; and

when a determination has been made that the second data set corresponds to bacteria, estimating the quantity of bacteria in the sample by comparing the first and second data sets.

Assignments (2)
LICENSE Recorded May 6, 2011
From: CANADIAN BLOOD SERVICES
To: LIGHTINTEGRA TECHNOLOGY INC.
Reel/Frame 026237/0240 →
ASSIGNMENT OF ASSIGNOR'S INTEREST Recorded Oct 7, 2009
From: MAURER, ELISABETH
To: CANADIAN BLOOD SERVICES
Reel/Frame 023340/0217 →