Identification of novel anti-persister activity for borrelia burgdorferi

A technology of Borrelia and spirochetes, which is applied in the field of identification of novel antiviral activity of Borrelia burgdorferi, and can solve the problems of inability to be screened by high-throughput, lengthy measurement, high background and the like

Pending Publication Date: 2017-09-08
张颖 +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

These assays are lengthy and cannot be used for high-throughput screening
Commonly used LIVE/DEAD BacLight assays have high background issues and c

Method used

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  • Identification of novel anti-persister activity for borrelia burgdorferi
  • Identification of novel anti-persister activity for borrelia burgdorferi
  • Identification of novel anti-persister activity for borrelia burgdorferi

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0126] Method for identifying novel antidepressant activity of Borrelia burgdorferi

[0127] Bacterial strains, media and cultures: Borrelia burgdorferi strain B31 was obtained from the American Type Tissue Collection. Borrelia burgdorferi was cultured in BSK-H medium (HiMedia Laboratories Pvt. Ltd.) containing 6% rabbit serum (Sigma-Aldrich, St. Louis, MO). All media were filter sterilized with a 0.2-μm filter. Cultures were incubated at 33°C in sterile 50-mL sealed conical tubes (BD Biosciences, San Diego, CA) without antibiotics. After 7 days, 100 μL of stationary phase B. burgdorferi culture (1×10 6 cells) was transferred to 96-well tissue culture microplates for drug selection.

[0128] Microscopy: Samples were examined on a Nikon Eclipse E800 microscope equipped with differential interference contrast (DIC) and epifluorescence illumination, and recorded with a Spot slider color camera. Cell proliferation assays were performed by direct counting using a bacterial cou...

Embodiment 2

[0164] Drug combination on Borrelia survival in vitro: eradication

[0165] Achieved with daptomycin, cefoperazone, and doxycycline

[0166] Materials and methods

[0167] Strains, media and cultures: Strains, media and cultures were obtained and used according to Example 1.

[0168] Antibiotics: Doxycycline (Dox), Amoxicillin (Amo), Cefoperazone (CefP), Clofazimine (Cfz), Miconazole (Mcz), Polymyxin B (Pmb), Sulfamethazine Oxazole (Smx), daptomycin (Dap), carbomycin (magnamycinA), nisin, carbenicillin, ofloxacin, tigecycline, hydroxychloroquine, rifampicin, and clarithromycin (Sigma-Aldrich) dissolved in a suitable solvent (Clinical and Laboratory Standards Institute, 2007). Antibiotic stocks were filter sterilized through a 0.2 μm filter, except for clofazimine, which was dissolved in DSMO (dimethyl sulfoxide) and not filtered. The stock was then stored at -20°C.

[0169] Microscopy: Samples were examined on a Nikon Eclipse E800 microscope equipped with differenti...

Embodiment 3

[0199] FDA-approved drug in circular form effective against Borrelia burgdorferi

[0200] Materials and methods

[0201] Strains, media and cultures: Borrelia burgdorferi strain B31 obtained from the American Type Culture Collection. Borrelia burgdorferi was cultured in BSK-H medium (HiMedia Laboratories Pvt. Ltd) with 6% rabbit serum (Sigma-Aldrich, Co). All media were filter sterilized through 0.2 μM filters. Cultures were incubated at 33°C without antibiotics in sterile 50 mL closed conical tubes (BD Biosciences, California, USA).

[0202] Induction of the Circular Body Form of Borrelia burgdorferi: To induce the Circular Body Form of Borrelia burgdorferi, Borrelia burgdorferi (1 x 10 5 spirochetes / ml) were cultured in BKS-H medium for 6 days without shaking. After 6 days of incubation, amoxicillin at a final concentration of 50 [mu]g / mL was added to the cultures for induction of the round body form. After induction at 33°C for 72 hours, the rounded form of Borrelia bu...

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Abstract

The presently disclosed subject matter provides methods, compositions, and kits for assessing the viability of bacteria from the Borrelia genus, assessing the antibiotic susceptibility of bacteria from the Borrelia genus, and identifying compounds with anti-persister activity for bacteria from the Borrelia genus. Methods for inhbiting the growth and/or survival of bacteria from the Borrelia genus and for treating Lyme disease in a subject are also are provided.

Description

[0001] CROSS-REFERENCE TO RELATED APPLICATIONS [0002] This application claims priority to US Provisional Patent Application No. 62 / 073,605, filed October 31, 2014, and US Provisional Patent Application No. 62 / 136,678, filed March 23, 2015, each of which All are incorporated herein by reference in their entirety. Background technique [0003] Lyme disease is a multisystem disease caused by the spirochete bacterium Borrelia burgdorferi (Meek et al., 1996; Stricker et al., 2011). The disease is transmitted by a tick vector, which can be transmitted by rodents, reptiles, birds and deer (Stricker et al., 2011; Radolf et al., 2012). In the United States, the number of Lyme disease cases has doubled over the past 15 years (Orloski et al., 2000; Bacon et al., 2008) and is estimated to be around 300,000 cases per year (Centers for Disease Control, 2014). Lyme disease is currently considered the most common tick-borne disease in the United States and Europe (Bacon et al., 2008; Arme...

Claims

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Application Information

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IPC IPC(8): C12Q1/06G01N21/25G01N21/64G01N33/50A61K31/00A61K38/00
CPCA61K31/366A61K31/4178A61K31/546A61K31/65A61K38/12A61K45/06C12Q1/04C12Q1/06C12Q1/18G01N21/6428G01N21/6458G01N2021/6441Y02A50/30A61K2300/00
Inventor 张颖冯杰
Owner 张颖
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