Method for detection of pathogenic organisms

a pathogenic organism and detection method technology, applied in the field of pathogenic organism detection, can solve the problems of time-consuming and difficult or impo differentiation between species of the same genus

Inactive Publication Date: 2014-01-16
HERRMANN BJORN +2
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Benefits of technology

[0008]The present invention solves the problem of differentiation between species within the same genus, such as mycobacteria and chlamydia. Furthermore, the present invention solves the problem of detecting pathogens which are hard and / or laborious to detect with conventional methods.

Problems solved by technology

This is reliable but time consuming, since slow-growing species such as Mycobacterium tuberculosis may need six to eight weeks to form a sufficiently large population.
a. The close relationship within microbial genuses, such as mycobacteria, has rendered differentiation between species of the same genus very difficult or impo

Method used

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  • Method for detection of pathogenic organisms
  • Method for detection of pathogenic organisms

Examples

Experimental program
Comparison scheme
Effect test

example 1

Mycobacteria

Materials and Methods

Bacterial Strains.

[0032]Mycobacteria used in this study are listed in Table 1 below. Clinical strains were already typed by 16S RNA gene sequencing in their respective institutes of provenance.

[0033]The RNase P RNA sequences for M. tuberculosis, M. bovis BCG and M. leprae were retrieved from the GenBank sequence database.

TABLE 1StrainSpeciesdescriptionProvenanceM. gastriATCC 15754ATCCM. kansasiiATCC 12748ATCCM. intracellulareD673Trudeau MycobacterialCollectionM. xenopiATCC 19276ATCCM. smegmatismc2 155Ref. 26M. aviumD702Trudeau MycobacterialCollectionM. marinumclinical strainResearch Centre BorstelM. fortuitumclinical strainResearch Centre BorstelM. malmoenseclinical strainResearch Centre BorstelM. paratuberculosis6783G-F Gerlach School ofVeterinary Medicine,Hanover GermanyM. gordoniaeclinical strainResearch Centre BorstelM. celatumclinical strainResearch Centre Borstel

PCR Amplification of the RNase P RNA Genes.

[0034]Based on the published sequences ...

example 2

Chlamydia

Materials and Methods

Bacterial Strains.

[0061]DNA of analysed organisms were released by standard Proteinase K treatment of cell culture grown organisms and phenol extracted or were provided as purified DNA-preparations (Table 2).

TABLE 2Strains, host of origin, references, sources and accession numbersStrainHost of originReferenceSource*Accession no.6BCT (VR-125T)†PsittacineGolub & Wagner (1947)StoreyAJ012169GDDuckIllner (1960)NADCNJ1TurkeyPage (1959)NADCWCCattlePage (1967)NADCVS225PsittacineNADC360DuckStoreyN352DuckRichmond et al. (1982)StoreyCal10HumanFrancis & Magill (1938)StoreyCP3 (VR-574)PigeonPage & Bankowski (1960)NADCM56 (VR-630)MuskratSpalatio et al. (1966)NADCB577T (VR-656T)Ovine, abortionPerez-Martinez & Storz (1985)DenamurAJ131092EBABovine, abortionPerez-Martinez & Storz (1985)NADCOSPOvineAndersen & Van Deusen (1988)NADC EAE / LxOvine, abortionStoreyAB7Ovine, abortionRodolakis et al. (1989)RodolakisOC1Ovine, conjunctivitisRodolakis et al. (1989)RodolakisAV1Bovine...

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Abstract

A method for detection of pathogenic organisms wherein the method includes differentiation between species. The method is especially suitable to detect and to diagnose infection by pathogenic organisms which are hard and / or laborious to detect with conventional methods. The method relies upon analysis of specific variable regions of the RNase P RNA gene, namely the P3 and / or P19 region(s).

Description

FIELD OF THE INVENTION[0001]The present invention relates to a method for detection of pathogenic organisms wherein the method includes differentiation between species. The method is especially suitable to detect and to diagnose infection by pathogenic organisms which are hard and / or laborious to detect with conventional methods. The method relies upon analysis of specific variable regions of the RNase P RNA gene.BACKGROUND OF THE INVENTION[0002]RNase P is an enzyme present in all living cells. It catalyses the removal of 5′ leader sequences from tRNA precursor molecules. In bacteria, RNase P consists of an RNA molecule of some 400 nt in length (11, 28) and a small (about 120 aa) protein (33). In the division bacteria, the RNA moiety has been shown to function as an efficient catalyst in vitro (12); hence at least in these organisms, RNase P is a ribozyme (an RNA molecule catalysing chemical reactions). Bacterial RNase P RNAs have been separated into two main structural classes. Typ...

Claims

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Application Information

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Patent Type & Authority Applications(United States)
IPC IPC(8): C12R1/32C12Q1/68C12N15/09C12Q1/689
CPCC12Q1/689
Inventor HERRMANN, BJORNKIRSEBOM, LEIFSTOLT, PELLE
Owner HERRMANN BJORN
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