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Biomarkers and uses thereof

A technology for biological and biological samples, applied in the field of identification and/or classification of microorganisms, which can solve the problems of lack of confidence in positive PCR results, lack of sensitivity and specificity, etc.

Pending Publication Date: 2020-03-24
MICROBIO PTY LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the problem with this technique is the lack of confidence in early positive PCR results in the absence of culture results or associated clinical or physical symptoms
Another problem with this technique is that, as with culture, there is a lack of sensitivity and specificity when attempting to detect small amounts of microbial nucleic acid in the background of host nucleic acid

Method used

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  • Biomarkers and uses thereof
  • Biomarkers and uses thereof
  • Biomarkers and uses thereof

Examples

Experimental program
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Effect test

Embodiment 1

[0490] The aim of the experiment was to identify 15 of the most common bacterial species frequently isolated from patients diagnosed with sepsis using the 16S rRNA SNP-HRM assay of the present invention.

[0491] Experimental procedure

[0492] In total, the following 15 bacterial species were tested: Acinetobacter calcoacetate; Enterobacter aerogenes; Enterobacter cloacae; Enterococcus faecalis; Enterococcus faecium; Escherichia coli; Klebsiella pneumoniae; Proteus mirabilis; ; Serratia marcescens; Staphylococcus aureus; Staphylococcus epidermidis; Streptococcus agalactiae; Streptococcus pneumoniae; and Streptococcus pyogenes.

[0493] Bacterial species were grown overnight in brain heart infusion at 37°C, and then genomic DNA was extracted from each isolate using the QIAgen DNeasy Blood and Tissue Kit (Qiagen, Australia).

[0494] 16S rDNA-SNP primers (SEQ ID NO: 16-35, manufactured by Sigma Aldrich, Australia) were designed to amplify a region containing seven SNPs desig...

Embodiment 2

[0513] To demonstrate the usefulness of the present invention, 200 patient samples with positive blood cultures were evaluated by standard clinical microbiology and the method of the present invention.

[0514]Standard clinical microbiological testing was performed by routine blood culture procedures using the BacTAlert system in the laboratory, followed by bacterial species identification using the MALDI biotyping method. This involves placing the BacTAlert blood culture bottle in an automated continuous monitoring incubator and incubating it for 5-7 days. Once the blood culture bottle was marked positive (at least 12 hours of incubation), the bottle was removed from the BacTAlert instrument, an aliquot of growth medium was removed, and subcultured on bacterial culture agar plates. Incubate the agar plates at 37°C for at least 4 hours, or until visible growth occurs. Thereafter, single bacterial colonies are plated on target plates and matrix solution is added. The plate wa...

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Abstract

The present invention generally relates to methods and agents for identifying and / or classifying microbes (especially bacteria), yeast organisms and filamentous fungi. In particular, the present invention concerns the discovery of unique single nucleotide polymorphisms (SNPs) in bacterial 16S ribosomal RNA (16S rRNA) and yeast organism and filamentous fungi 18S ribosomal RNA, and methods of classifying and / or identifying bacteria, yeast organisms and / or filamentous fungi in a sample based on the presence of one or more of those SNPs. The present invention also concerns probes, primers and kitsthat are useful in those methods.

Description

technical field [0001] The present invention generally relates to methods and reagents for identifying and / or classifying microorganisms. The methods and reagents are based on the detection of polymorphisms within the 16S ribosomal RNA gene or gene product of bacteria and within the 18S ribosomal RNA gene or gene product of yeast organisms and filamentous fungi. The invention also features methods for treating bacterial, yeast organism, or filamentous fungal infections based on the diagnostic methods of the invention. Background technique [0002] It is highly desirable to quickly and accurately identify or classify microorganisms (such as bacteria), yeast organisms, and filamentous fungi in a sample. [0003] First, such infections can be diagnosed quickly and accurately by allowing early implementation of effective treatment or therapy, which can make the difference between life and death for a patient. [0004] Rapid and accurate identification or classification can aid...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/689C12Q1/6895C12Q1/04
CPCC12Q1/04C12Q1/689C12Q1/6895C12Q2600/156C12Q1/686C12Q2600/112
Inventor F·惠更斯
Owner MICROBIO PTY LTD
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