Method for detecting gram-positive bacteria
AssigneeHain Lifescience GmbH
InventorMichael Weiznegger
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Abstract
The present invention relates to a method for detecting and/or identifying Gram-positive bacteria with high GC content, in particular mycobacteria. The method comprises a nucleic acid amplification reaction and a subsequent hybridization reaction with suitable primers and probes. The invention furthermore relates to oligonucleotides, compositions, solid phases and kits which may be used for carrying out the method of the invention.
Claims23
A method for detecting and/or identifying Gram-positive bacteria with high GC content, which comprises
- a) subjecting a sample composition to a nucleic acid amplification reaction, with there being a first primer selected from the group consisting of the primers having the sequences SEQ ID NO. 1, SEQ ID NO. 3, SEQ ID NO. 5, SEQ ID NO. 7 and SEQ ID NO. 9 and there being a second primer selected from the group consisting of the primers having the sequences SEQ ID NO. 2, SEQ ID NO. 4, SEQ ID NO. 6, SEQ ID NO. 8 and SEQ ID NO. 10;
- b) hybridizing a composition comprising the amplification product of step a) or a part thereof with one or more probes selected from the group consisting of oligonucleotides having the sequences SEQ ID NO. 11 to 42 and sequences complementary thereto;
- and
- c) determining the degree of hybridization.
The method as claimed in either of the preceding claims, wherein any of the primer pairs having the sequences
- SEQ ID NO. 1/SEQ ID NO. 2 (primer pair 1),
- SEQ ID NO. 3/SEQ ID NO. 4 (primer pair 2),
- SEQ ID NO. 5/SEQ ID NO. 6 (primer pair 3),
- SEQ ID NO. 7/SEQ ID NO. 8 (primer pair 4) and
- SEQ ID NO. 9/SEQ ID NO. 10 (primer pair 5)
- is present in the nucleic acid amplification reaction.
The method as claimed in any of the preceding claims, wherein at least one probe or one primer is labeled.
An oligonucleotide having a sequence selected from the group consisting of the sequences SEQ ID NO. 1 to 42 and sequences complementary to the sequences SEQ ID NO. 11 to 42.
A solid phase on which the oligonucleotides having the sequences SEQ ID NO. 11 to 42 or a sequence complementary thereto are immobilized, the various oligonucleotides being spatially separated from one another.
The use of one or more of the oligonucleotides having the sequences SEQ ID NO. 1 to 10 as primers in a nucleic acid amplification reaction.
The use of one or more of the oligonucleotides having the sequences SEQ ID NO. 11 to 42 or sequences complementary thereto as probe in a hybridization reaction.
Dependencies are inferred from the claim text.