System for detecting bacteria in blood, blood products, and fluids of tissues

a tissue and blood technology, applied in the field of tissue and blood products, can solve the problems of inability to use donor blood and blood products or donor organs, and inability to quickly and efficiently detect the presence of contaminating bacteria in blood or blood products, etc., to achieve rapid propagation rate, short shelf life, and increase the amount of contaminating bacteria presen

Inactive Publication Date: 2010-05-27
VERAX BIOMEDICAL
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Thus, if blood contaminated with a clinically relevant amount of bacteria were to be transfused to a recipient patient, the recipient patient might suffer complications, particularly since transfusions of blood and / or blood products are often performed when the patient is undergoing major surgery, or is otherwise vulnerable.
Despite the need for a safe, microbe-free supply of blood or a blood product, no rapid, efficient method exists for detecting the presence of contaminating bacteria in blood or a blood product.
Although these culture-based blood testing techniques are useful for determining the particular type of bacteria that is infecting a patient's blood, the length of the time required to perform these techniques makes them impractical to use to test donor blood and blood products or donor organs.
Thus, there may be no time to test the donated blood (or blood product) or organ with a culture-based technique before the donated blood or organ is needed for use.
In addition, donated organs and blood or blood products have a relatively short shelf-life due not only to a loss of function of the donated material, but also to an increase in amount of any contaminating bacteria present.
Moreover, these culture-based blood testing techniques take too long to routinely screen platelets, which have a short shelf-life (approximately 5 days post-donation), for transfusion use.
(Transfusion 34(9): 750-755 (1994)) teaches another technique for detecting a particular contaminating bacteria in blood using labelled nucleic acid probes to hybridize to the genetic material of potential contaminants The probes used in these studies, however, are very limited in the number of micro-organisms that can be detected and, unfortunately, no commercially viable test has emerged from this technology.
Given their complexity, these techniques are too labor-intensive and too time-costly to be routinely used to screen blood or blood products for bacterial contamination.

Method used

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  • System for detecting bacteria in blood, blood products, and fluids of tissues
  • System for detecting bacteria in blood, blood products, and fluids of tissues
  • System for detecting bacteria in blood, blood products, and fluids of tissues

Examples

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example i

[0142]A bacterial detection method of the invention can follow a conventional particle-based immunoassay format. In this approach, binding agents that specifically bind to an antigenic region on the bacterial cell wall can be utilized as immunological targets and can form the basis for detecting the bacteria in the blood products. A schematic of the device used in this example is shown on FIGS. 3A-3D. In one embodiment, the binding agents that specifically bind to Gram-positive bacteria, Gram-negative bacteria, or both, are pooled together, and then roughly divided, such that the two populations of binding agents each comprises a mixture of binding agents. One population is bound to the membrane at the base of the device shown in FIGS. 3A-3D. The second population is used to coat brightly dyed latex particles (i.e., the latex particles coated with the binding agent have been impregnated with a dye so that they are intensely colored).

[0143]In another embodiment, the same binding agen...

example ii

[0149]In another embodiment of the method described in Example I, the latex-bound binding agents are binding agents that specifically recognize either Gram-negative bacteria or Gram-positive bacteria, or both. However, each of these anti-bacteria binding agents has an epitope that is common to all binding agents. In this example, all of the anti-bacteria binding agents are murine antibodies. The membrane bound binding agents do not specifically bind bacteria; rather, they specifically bind the murine determinants on the constant region of the murine anti-bacteria antibodies. These anti-mouse binding agents are called secondary binding agents, because they do not specifically bind the bacteria, but rather, specifically bind the binding agent that is specifically bound to the bacteria.

example iii

[0150]An agglutination assay is yet another method to detect clinically relevant amounts of bacteria in blood, a blood product, or fluid from a donor tissue. In this example of an agglutination assay, latex particles coated with binding agents that specifically bind to bacterial species are employed. These latex particles agglutinate in the presence of clinically relevant amounts of bacterial species. A positive reaction is indicated by the development of an agglutinated pattern. In a negative reaction the latex does not agglutinate and the milky appearance remains substantially unchanged.

[0151]In this method, a test device such as that shown on FIGS. 4A-4D is employed. A sample extracted under sterile conditions from a donor blood or blood product is applied to the test device. Alternatively, a sample extracted from a fluid in which a donor organ or tissue is stored may be applied to the test device. The sample may be tested straight (i.e., undiluted or untreated), diluted with a b...

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Abstract

The invention provides methods for screening for the presence of a clinically relevant amount of bacteria in donor blood or a blood product from a donor mammal, particularly blood or a blood product that will be transferred from the donor mammal to a recipient mammal. The method comprises contacting a sample of the donor blood or a blood product with a set of binding agents that comprises binding agents that specifically bind to Gram-negative bacterial antigen and / or binding agents that specifically bind to Gram-positive bacterial antigen, and determining binding of the set of binding agents to the sample, wherein binding indicates the presence of a clinically relevant amount of Gram-positive bacteria and / or Gram-negative bacteria in the donor blood or blood product and no binding indicates the absence of a clinically relevant amount of Gram-positive bacteria and / or Gram-negative bacteria in the donor blood or blood product.The invention further provides methods and kits for screening for the presence of a clinically relevant amount of Gram-positive bacteria, Gram-negative bacteria, or both Gram-positive and Gram-negative bacteria in a donor tissue by screening the fluid in which the donor tissue is stored.

Description

CROSS-REFERENCE TO RELATED APPLICATIONS[0001]This application claims benefit from U.S. Provisional Patent Application No. 60 / 144,442 filed Jul. 16, 1999, the entire contents of which is hereby incorporated by reference.BACKGROUND OF THE INVENTION[0002]1. Field of the Invention[0003]The invention relates to the screening of blood or a blood product (including whole blood, hematopoietic stem cells, leukocytes, plasma, serum, red blood cells, and platelets) or a donor tissue for the presence of a clinically relevant amount of bacteria. More particularly, the invention relates to the screening for the presence of a clinically relevant amount of contaminating bacteria in blood and blood products or donor tissue that will be used for transfusion or transplantation.[0004]2. Summary of the Related Art[0005]Transfusion of blood and blood products is a therapeutically important aspect of patient care. Transplantation of donor tissues and organs is likewise therapeutically important. The absen...

Claims

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Application Information

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Patent Type & Authority Applications(United States)
IPC IPC(8): G01N33/543G01N33/569
CPCG01N33/56911
Inventor GOODNOW, TIMOTHY T.
Owner VERAX BIOMEDICAL
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