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Multiplex assay detection of pathogenic organisms

A pathogenic and biological technology used in the detection of pathogenic microorganisms and the detection of infections caused by pathogenic organisms in clinical samples

Inactive Publication Date: 2016-02-24
F HOFFMANN LA ROCHE & CO AG
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0030] The above disclosed problems are solved by the methods and compounds disclosed and claimed in this application

Method used

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  • Multiplex assay detection of pathogenic organisms
  • Multiplex assay detection of pathogenic organisms
  • Multiplex assay detection of pathogenic organisms

Examples

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Embodiment

[0155] 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.

[0156] sample

[0157] Sections containing 18SrRNA and 23sRNA from E. 10 of the ITS-1 region of the plasmid DNA 3 、10 2 and 10 copies. An internal control plasmid was added to the completed mastermix (see above).

[0158] hardware / software

[0159] 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 consisted of 4 photohybrids instead of 3 and could detect fluorescence emissions at 610 nm, 640 nm, 670 nm and 705 nm.

[0160] Reagent

[0161] All oligonucleotides mentioned he...

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Abstract

The present invention relates to multiplex assay detection of pathogenic organisms. The present invention is directed to a method for identification of a pathogenic organism from a predetermined group of pathogens, comprising (i)isolating a clinical sample containing at least partially purified nucleic acid, (ii) subjecting at least a first aliquot of said clinical specimen to at least one amplification and detection reaction in one reaction vessel comprising (iia) an amplification step using at least a first set of amplification primers capable of amplifying a preselected nucleic acid sequence region from several or all members of said predetermined group of pathogens, (iib) a detection step using at least 2, 3 or multiple hybridization reagents, said reagents together being capable of specifically detecting a pre-selected nucleic acid sequence region from all members of said group of pathogens, said detection step (iib); comprising steps monitoring hybridization of each of said hybridization reagents at a pre-selected temperature, said hybridization being indicative for at least the genus of said pathogen present in the sample, and monitoring temperature dependence of hybridization, said temperature dependence being indicative for at least the species of said pathogen, and (iii) determining whether said amplification and detection reaction is indicative for the presence of a specific member of said pre-selected group of pathogens.

Description

[0001] This application is a divisional application of the invention patent application titled "Multiple analysis and detection of pathogenic organisms" with the international application date of December 2, 2003 entering China with the international application PCT / EP2003 / 013530 and the application number 200380105073.7. technical field [0002] 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 [0003] 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 wit...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68C12Q1/04C12P19/34
CPCC12Q1/6816C12Q1/686C12Q2531/113C12Q2545/101C12Q1/6851C12Q1/689
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
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