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Multiplex assay detection of pathogenic organisms

A pathogenic and biological technology, applied in the field of detecting pathogenic microorganisms and detecting infections caused by pathogenic organisms in clinical samples

Inactive Publication Date: 2006-01-11
F HOFFMANN LA ROCHE & CO AG
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0029] The above disclosed problems are solved by the methods and compounds disclosed and claimed in this application

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment

[0154] The following examples, references and sequence listing are provided to aid in the understanding of the present invention, the true scope of which is set forth in the appended claims. It is understood that modifications may be made in the steps presented without departing from the spirit of the invention.

[0155] sample

[0156] Sections containing 18S rRNA and 23 sRNA from E. faecium, E. faecalis, S. aureus, S. epidermidis, or Pseudomonas aeruginosa were analyzed in duplicate in the absence or presence of a background of 5 μg human genomic DNA 10 of the plasmid DNA between the ITS-1 region 3 、10 2 and 10 copies. An internal control plasmid was added to the completed mastermix (see above).

[0157] hardware / software

[0158] Use LightCycler TM Instrumentation (Roche Diagnostics GmbH, Germany). The commercially available instrument was adapted so that the rotor was adapted to accommodate 100 μl capillaries. The instrument's fluorometer was modified so that it co...

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PUM

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Abstract

The invention relates to a method for analyzing the presence of a bacterial pathogen in a clinical sample comprising the steps of (i) at least partially isolating nucleic acid from the sample, (ii) putting at least the first divide sample of the clinical sample into a reaction vessel to carry out at least one amplification and detection reaction wherein, the reaction comprises iia) the step of amplification through at least one first amplification primer which can amplify the nucleotide sequence area selected from some or all of the members of the bacterial pathogen group; iib) the step of detection through at least two, three or more half-bred reagents which can detect the nucleotide sequence area selected from all of the members of the bacterial pathogen group, wherein the step of detection iib) comprises the steps as follow: monitoring the half-bred of each one of half-bred reagents at a preset temperature, which at least indicts the property of the bacterial pathogen in the sample; monitoring the temperature dependency of the half-bred which at least indicts the type of the bacterial pathogen; and (iii) determining whether the amplification and detection reaction indicts the existence of the special members of the preset group of the bacterial pathogen or not.

Description

[0001] The invention relates to the technical field of detecting pathogenic microorganisms. More specifically, the present invention relates to the field of detection of infections by pathogenic organisms in clinical samples by amplifying and detecting specific nucleic acid sequences from said pathogenic organisms. Background technique [0002] Infections by pathogenic bacteria, especially those that cause sepsis, mainly occur in the intensive care unit (ICU) of a hospital and are serious. The bacteria infecting patients is in most cases unknown and cannot be determined from symptoms. Each type of bacteria requires a different therapy with specific antibiotics. Currently, in routine diagnostics, use includes culturing bacteria (if present) from samples of blood or other body fluids to detect pathogenic bacteria, especially Gram-positive bacteria. This culture is maintained under conditions favorable for bacterial growth for about 3 days. During this period, the number of ba...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68C12Q1/04C12Q1/14G01N33/542G01N33/58
CPCC12Q1/14C12Q1/04C12Q1/6851G01N2333/315G01N2333/31C12Q1/689
Inventor G·哈伯豪森T·埃姆里希G·萨纳M·莫茨科G·施米茨-阿赫吉安
Owner F HOFFMANN LA ROCHE & CO AG