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Methods and compositions to prevent microbial infection

A technology of microorganisms and target microorganisms, applied in the direction of microorganisms, biochemical equipment and methods, drug delivery, etc., can solve the problems of variable recurrence of pathogenic microorganisms, inability to induce β-lactamase, low toxicity, etc.

Pending Publication Date: 2020-11-10
比奥普来克斯有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

502a is a low-virulence coagulase-positive strain of Staphylococcus aureus that is susceptible to penicillin and fails to induce β-lactamase
[0044] Prior art methods involving the suppression (decolonization) and replacement (recolonization) of the original pathogenic microorganisms with new microorganisms may result in variable recurrence of pathogenic microorganisms depending on the specific method

Method used

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  • Methods and compositions to prevent microbial infection
  • Methods and compositions to prevent microbial infection
  • Methods and compositions to prevent microbial infection

Examples

Experimental program
Comparison scheme
Effect test

example 1

[0672] Example 1. Field study - Exclusive niche using benign microbes:

[0673] Clinical Research - Inhibition and Replacement

[0674] A clinical study was designed to identify MRSA positive subjects, suppress MRSA strains, replace MRSA by administering Bioplx-01 (ie, MSSA 502a) and periodically retest subjects for recurrence of MRSA. The study population was mainly medical staff and medical students from the Meerut region. Subjects without symptoms were included in the study.

[0675] This was a 'proof-of-principle' study using largely unrefined materials and methods - any result greater than a 55% relapse-free rate would be considered an indicator of the potential efficacy of these methods. Any result of a recurrence-free rate of 80% or higher would be considered a strong indicator of the current technical strength of the method.

[0676] Study objectives and primary endpoints:

[0677] 1) To determine the prevalence of asymptomatic S. aureus and MRSA in the general pop...

example 2

[0727] Example 2. Selection of one or more inducible promoters

[0728] In this example, promoter candidates were evaluated. Fold induction and basal expression of six promoter candidates in MSSA strain BioPlx-01 were assessed by incubation with human whole blood and serum. Expression was normalized to a housekeeping gene (gyrB) and compared to that in cells grown logarithmically in liquid tryptic soy broth (TSB) medium.

[0729] BioPlx-01 was grown to mid-log phase (2OD / mL), then washed extensively and transferred to freshly collected serum and heparinized blood from donor TK.

[0730] Samples were incubated at 125 rpm in slowly stirred vented flasks; and samples were removed for RNA isolation at 15 min, 45 min or 75 min at 37°C. Collected bacteria were washed and RNA was extracted using Qiagen Allprep kit, eluted and frozen. Coding DNA (cDNA) was prepared from RNA and expression of target genes was assessed by real-time PCR (Taqman) in an ABI 7500 Fast instrument.

[073...

example 3

[0748] Example 3. Selection of one or more death genes

[0749] In this example, cell death gene candidates are evaluated to produce synthetic microorganisms having at least one molecular modification comprising a first cell death operably linked to a first regulatory region comprising a first inducible promoter Gene. The relative potency of death genes is unknown. Because of leaky expression, the best-looking death gene is not necessarily the most efficient gene. The diversity of mechanisms of action may result in the synergistic killing effect of combinations of two or more death genes. Candidate death genes include: SprA1: membrane rupture; smal: genome disruption; and rsaE: block central metabolism. Various combinations of death genes are shown in Table 14. These plasmids were created and sequenced to test the P leuA and P hlgA Driven KS variants.

[0750] Table 14. Death gene KS constructs

[0751]

[0752] The death gene is commercially available (Atum), as is...

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Abstract

Methods and compositions are provided for durably influencing microbiological ecosystems (microbiomes) in a subject in order to prevent infection and reduce recurrence of infection by microorganisms.In some embodiments, compositions and methods are provided for the creation and use of molecularly-modified bacterial strains with the potential to prevent a variety of microorganism infections.

Description

[0001] Cross References to Related Applications [0002] This application was filed as a PCT International Patent Application on December 4, 2018, and claims priority to U.S. Provisional Application No. 62 / 594,943, filed December 5, 2017, which is hereby incorporated by reference in its entirety. [0003] sequence listing [0004] This application contains a Sequence Listing in electronic format as a file entitled "Sequence-Listing" created on December 3, 2018, and said file is 128,065 kilobytes (KB) in size. The contents of the txt file "Sequence-Listing" are incorporated herein by reference. technical field [0005] Methods and compositions are provided for durably affecting the microbial ecosystem (microbiome) in a subject to combat infection and reduce the recurrence of infection by undesirable microorganisms through decolonization and permanent replacement with synthetic microorganisms. Provided are synthetic microorganisms that can durably replace undesirable microo...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/21A61K35/74A61K35/741A61K45/06A61P31/04
CPCC12N15/77C12N15/78C12N15/74A61K9/0014A61P31/04C12N9/16C07K14/245C07K14/31C12Y301/21004A61K45/06A61K35/741A61K2300/00A61K35/74C12N2840/002A61K2035/115A61K2035/11C12N2820/55C12N2820/002C12N2830/55C12N2840/55C12N1/20C12N15/63C12N15/64C12N15/09
Inventor 蒂莫西·W·斯塔兹尔托德·D·图尔纳拉维·S·V·斯塔兹尔
Owner 比奥普来克斯有限公司
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