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Nmr methods and systems for the rapid detection of tick-borne pathogens

a technology of nmr methods and ticks, applied in the direction of instruments, biochemistry apparatus and processes, analysis using nuclear magnetic resonance, etc., can solve the problems of chronic lyme disease with recurring muscle and joint pain, untreated tick-borne diseases can have serious health consequences, and difficult to differentially diagnose tick-borne diseases

Pending Publication Date: 2022-07-21
T2 BIOSYST
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Benefits of technology

The invention is about methods for detecting tick-borne pathogens, such as Borrelia burgdorferi, in biological samples. The methods involve amplifying specific nucleic acids and detecting them using magnetic particles and an RF coil. The invention can detect the presence of multiple species of Borrelia, making it useful for diagnosing and treating tick-transmitted diseases like Lyme disease. The methods have high sensitivity, with the ability to detect a small number of cells of the pathogen in a sample. The invention also includes a device for performing the detection method.

Problems solved by technology

Tick-transmitted diseases can be difficult to differentially diagnose, especially in their early phases, in part because these diseases tend to have similar symptoms, which may be variable between patients.
However, untreated tick-transmitted diseases can have serious health consequences.
Further complications can arise as the disease progresses, which manifest in nerve damage, brain inflammation, and arthritis (stage Ill), culminating in chronic Lyme disease with recurring muscle and joint pain, cognitive defects, and chronic fatigue.
The difficulty in diagnosing tick-transmitted diseases is compounded by difficulties in detecting the causative pathogens, which may be present in low titer in the infected subject's tissue (e.g, blood, cerebrospinal fluid, urine, and / or synovial fluid, depending on the pathogen and the progression of the disease).
Available diagnostic tests for these diseases generally have drawbacks, including being time-consuming and / or having relatively low sensitivity.
For example, diagnosis of Lyme disease currently involves a two-tiered serological test, with none of the available nucleic acid-based tests (e.g., polymerase chain reaction (PCR)) being FDA-approved, in part due to sensitivity limitations.
This sample preparation is time consuming, and also can lower the sensitivity of the assay due to loss of analyte.
Additionally, current tests are focused on detecting single pathogens, which can further delay diagnosis if the incorrect test is selected.

Method used

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  • Nmr methods and systems for the rapid detection of tick-borne pathogens
  • Nmr methods and systems for the rapid detection of tick-borne pathogens
  • Nmr methods and systems for the rapid detection of tick-borne pathogens

Examples

Experimental program
Comparison scheme
Effect test

example 1

on Blood Assay

[0314]One assay of the present invention is a combination assay that detects Borrelia burgdorferi sensu lato (Bbsl)-specific antibodies (IgM and IgG) and Bbsl-specific nucleic acid targets in whole blood specimens. To test a blood sample for the presence of a Bbsl species, for example during diagnosis of Lyme disease, a single blood sample drawn from the subject may be subjected to simple fractionation by high speed centrifugation. Antibodies expressed by a subject in response to Bbsl infection may be present in serum, which is the supernatant produced by the high-speed centrifugation. These antibodies can be detected by a serological test, including those known in the art, or using methods of the invention as described herein. In contrast, pathogen-specific nucleic acids, if present, will be concentrated in the pellet, which may be lysed to form a lysate for detection. The detection may involve an amplification step, such as PCR.

example 2

Comprehensive Detection of Tick-Borne Pathogens

[0315]This example describes an exemplary panel of the invention. Detection of this panel enables rapid and accurate differential diagnosis of tick-transmitted diseases, which often manifest with similar symptoms. A patient presenting with symptoms consistent with a tick-transmitted disease may be tested for the panel, which may be performed in a multiplexed assay.

[0316]The panel includes two sub-panels (see FIG. 1). The first sub-panel, termed “Lyme panel,” targets Bbsl and pan-Borrelia species, which are extracellular bacteria that are viable in blood and that can also disseminate into and establish themselves in non-blood tissues, including cerebrospinal fluid, urine, and synovial fluid. Titer levels in the different tissues, including blood, vary depending on the stage of the disease. Using the methods of the invention, this panel can be used to rapidly and sensitively diagnose Lyme disease in both the acute (stage I) and later stag...

example 3

for Detecting Tick-Borne or Other Pathogens Using the T2Dx® Instrument

[0320]The methods described herein may be performed using a T2Dx® instrument (T2 Biosystems, Lexington, Mass.). A sample obtained from a subject (e.g., a 1.7-2 mL whole blood sample from a human suspected of being infected with a tick-borne pathogen) is inserted into the device, for example, as described in WO 2012 / 054639. If the sample is a whole blood sample, the next step typically includes blood cell lysis and concentration, for instance, by (a) by mixing the whole blood sample with an erythrocyte lysis agent solution to produce disrupted red blood cells, (b) centrifuging the sample to form a supernatant and a pellet, discarding some or all of the supernatant, and optionally washing the pellet (for example, by resuspending the pellet in volume of a buffer, and centrifuging the pellet), and (c) lysing cells in the pellet (which may include white blood cells and / or pathogen cells) to form a lysate. The lysis may...

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PUM

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Abstract

The invention features methods, systems, and panels for rapid detection of tick-borne pathogens in a sample (including Borrelia spp. such as B. burgdorferi, B. afzelii, and B. garinii) and for diagnosis and monitoring of tick-transmitted diseases, including Lyme disease, Rocky Mountain spotted fever, Q-fever, babesiosis, ehrlichiosis, tularemia, and anaplasmosis.

Description

FIELD OF THE INVENTION[0001]The invention features methods, panels, and systems for detecting tick-borne pathogens and diagnosing tick-transmitted diseases.BACKGROUND OF THE INVENTION[0002]A number of diseases are transmitted by ectoparasites such as ticks, including Lyme disease, Rocky Mountain spotted fever, Q-fever, babesiosis, ehrlichiosis, tularemia, and anaplasmosis, which are caused by the tick-borne pathogens Borrelia burgdorferi sensu lato (Bbsl; B. burgdorferi, afzelii, and garinii), Rickettsia rickettsii, Coxiella burnetii, Babesia microti, Ehrlichia chaffeensis, Francisella tularensis, and Anaplasma phagocytophilum, respectively. Tick-transmitted diseases can be difficult to differentially diagnose, especially in their early phases, in part because these diseases tend to have similar symptoms, which may be variable between patients. However, untreated tick-transmitted diseases can have serious health consequences. As an example, for Lyme disease, following the initial, l...

Claims

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Application Information

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Patent Type & Authority Applications(United States)
IPC IPC(8): C12Q1/689C12N15/10G01N24/08
CPCC12Q1/689C12N15/1013G01N24/08C12Q2600/112G01N2800/26Y02A50/30C12Q2600/124G01N2333/20C12Q2600/156
Inventor THOMANN, ULRICH HANSNEELY, LORI ANNEDHANDA, RAHUL KRISHANSWEENEY, FRÉDÉRICLOWERY, JR., THOMAS JAYTOWNSEND, JESSICA ANNSNYDER, JESSICA LEEGIESE, HEIDI SUSANNEBANDOSKI-GRALINSKI, CHERYL ANNLOGAN, DANIELLA LYNN
Owner T2 BIOSYST
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