IP Library Granted Patent US 7,169,555
Granted Patent B2
US 7,169,555 · App. 10/168,639 · Granted Jan 30, 2007

Method for indentifying microbial DNA or RNA

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Quick Facts
Patent No.
US 7,169,555
App. No.
10/168,639
Granted
Jan 30, 2007
Kind
B2
Abstract

The invention relates to a method for quickly detecting bacterial DNA. The sample containing bacterial DNA is provided and treated according to a PCR method. The DNA copies which are multiplied by means of PCR are detected.

Claims (10)

1. A method for identifying microbial DNA or RNA comprising:

(a) providing a sample containing microbial DNA or RNA;

(b) enriching ribosomal DNA or RNA in the sample by a PCR method wherein the distance between two primers is between 100 and 250 base pairs on a template to be amplified with the primers, the primers containing 16 to 25 nucleotides, one of said primers being selected from the group consisting of PLK2H (SEQ ID NO: 47), REViFL (SEQ ID NO: 2) and iFL (SEQ ID NO: 1);

(c) preliminarily identifying at least one microorganism on the basis of the melting temperature of the resultant amplification product from step (b);

(d) adding at least one labeled oligonucleotide probe to said enriched ribosomal DNA or RNA whereby temperature-dependent hybridization takes place, wherein the probe is selected from the group consisting of ISN2 (SEQ ID NO: 43), ISN2* (SEQ ID NO: 44), ISN (SEQ ID NO: 34), ISP2 (SEQ ID NO: 46), ISP2* (SEQ ID NO: 45) and ISP (SEQ ID NO: 33);

(e) obtaining a melting curve for the resultant probe-enriched ribosomal DNA or RNA hybridization products; and (f) identifying said at least one microorganism from its melting curve.

2. The method as claimed in claim 1 , wherein the PCR method is a real-time PCR method and a quantity of the microbial DNA or RNA in an initial sample is determined.

3. The method as claimed in claim 1 , wherein the sample is a body fluid, a smear or a tissue.

4. The method as claimed in claim 1 , wherein the labeled oligonucleotide comprises a fluorescence transfer coupling dye-labeled probe; and a lower primer is provided which includes a dye which is capable of being transferred and which is suitable for fluorescence transfer coupling, the probe and primer being such that they bind to adjacent regions of a sequence to be amplified.

5. The method as claimed in claim 1 , wherein the method is for diagnosing a bacterial infection which is sepsis.