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Indicator bacteriophage for selecting and monitoring for efficacy of therapeutics and methods for using same

A phage and indicator technology, which is applied in the field of selection and monitoring of indicator phages for therapeutic efficacy and its use, can solve problems such as inappropriateness and cost

Pending Publication Date: 2021-02-02
LAB OF AMERICA HLDG
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In practice, most high-sensitivity methods utilize overnight incubations, and the entire process takes about 24 hours
Due to the time required for incubation, these methods can take up to three days, depending on the organism to be identified and the source of the sample
This lag time is often inappropriate because contaminated food, water (or other products) may have entered livestock or humans

Method used

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  • Indicator bacteriophage for selecting and monitoring for efficacy of therapeutics and methods for using same
  • Indicator bacteriophage for selecting and monitoring for efficacy of therapeutics and methods for using same
  • Indicator bacteriophage for selecting and monitoring for efficacy of therapeutics and methods for using same

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0196] Example 1. Bacterial detection using the S. aureus-specific phage NanoLuc indicator phage after incubating the sample in culture medium

[0197] Samples of methicillin-resistant S. aureus (MRSA) were made susceptible by adding 135 μl of samples containing S. aureus to the rich medium. Rich medium contains a sub-inhibitory antibiotic (cefoxitin) to allow specific enrichment and induction of MRSA. Incubate samples in rich medium for 1-2 hours. After the samples were sensitized, additional cefoxitin was added to the medium and the samples were incubated for an additional 2 hours. S. aureus-specific phage NanoLuc indicator phage was added to the samples and incubated for 2 hours. Add lysis buffer and reagent. After 5 minutes of incubation, use 96 instruments for bioluminescence measurements. A signal / background ratio ≥ 750 RLU indicates positive detection of S. aureus. A signal / background ratio of <750 RLU indicates that the sample was negative for S. aureus.

[0...

Embodiment 2

[0211] Example 2. Use of Staphylococcus spp.-specific phage after incubation of samples in culture medium Indicator phages for diagnosis and monitoring of Staphylococcus infections

[0212] Clinical samples are obtained from patients. Analyzing clinical samples with Staphylococcus-specific phage Indicator phage were incubated for 2 hours. After incubation, add lysis buffer and reagent. with lysis buffer and After reagent incubation for 5 minutes, use The 96 instrument measures bioluminescence to determine the presence of Staphylococcus species in clinical samples. A signal / background ratio > 750 RLU indicates positive detection of Staphylococcus. A signal / background ratio <750 RLU indicates that the sample is negative for Staphylococcus.

[0213]Patients with samples positive for Staphylococcus are treated with a therapeutic phage specific for Staphylococcus. A second clinical sample was obtained from the patient 72 hours after treatment with the therapeutic phag...

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Abstract

Disclosed herein are methods and systems for detection of microorganisms in a sample and the utilization of such methods for selecting and monitoring therapies. The specificity of indicator bacteriophage, such as Staphylococcus-specific bacteriophage, allows detection of a specific microorganism, such as Staphylococcus and the indicator signal may be amplified to optimize assay sensitivity.

Description

[0001] Cross References to Related Applications [0002] This application claims priority to U.S. Provisional Application No. 62 / 661,739, filed April 24, 2018. The disclosures of US Application Nos. 13 / 773,339, 14 / 625,481, 15 / 263,619, and 15 / 409,258 and US Provisional Application 62 / 661,739 are hereby incorporated by reference in their entirety. technical field [0003] The present invention relates to compositions, methods, systems and kits for detecting microorganisms using infectious agents. Background technique [0004] There is a strong interest in improving the speed and sensitivity of detection of bacteria, viruses and other microorganisms in biological, food, water and clinical samples. Microbial pathogens can cause massive disease in humans and livestock, as well as huge economic losses. Furthermore, given the outbreak of life-threatening or fatal illnesses caused by consumption of food contaminated with certain microorganisms (e.g., Staphylococcus spp.), the Food...

Claims

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Application Information

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IPC IPC(8): C12N15/65C12N15/75C12N7/00C12Q1/14C12Q1/70A61K35/76
CPCC12N7/00C12N15/65C12N2800/202C12N2840/203C12Q1/14C12N2795/10143C12N2795/10151C12N2795/10131C12N15/74C12Q1/689C12Q2600/106C12Q2600/158C12Q1/66
Inventor J·S·吉尔S·埃里克森M·M·B·恩古彦D·L·安德森
Owner LAB OF AMERICA HLDG