Detection, identification and differentiation of Staphylococcus using a hybridization assay

A method for the detection and/or identification of Staphylococcus aureus in a sample, comprising the steps of

• (i) hybridizing the polynucleic acids of the sample with at least two polynucleotide probes that hybridize to a target sequence within 25 nucleotides of each other, wherein the at least two polynucleotide probes are SEQ ID NO:17 and SEQ ID NO:19, and wherein the target sequence comprises at least 50 consecutive nucleotides from a nucleic acid molecule of
• SEQ ID NO:1 or
• SEQ ID NO:
• 2 or
• a Staphylococcus aureus sequence homologous to SEQ ID NO:1, or
• a Staphylococcus aureus sequence homologous to SEQ ID NO:
• 2, or
• the RNA form of SEQ ID NO:
• 1 wherein T is replaced by U, or
• the RNA form of SEQ ID NO 2 wherein T is replaced by U, or
• the RNA form of said Staphylococcus aureus specific homologue of SEQ ID NO:
• 1 wherein T is replaced by U, or
• the RNA form of said Staphylococcus aureus specific homologue of SEQ ID NO:
• 2 wherein T is replaced by U, or
• the sequence complementary to SEQ ID NO:1
• the sequence complementary to SEQ ID NO:2
• the sequence complementary to the Staphylococcus aureus sequence homologous to SEQ ID NO:1, or
• the sequence complementary to the Staphylococcus aureus sequence homologous to SEQ ID NO:2, or
• the sequence complementary to the RNA form of said SEQ ID NO:1 wherein T is replaced by U, or
• the sequence complementary to the RNA form of said SEQ ID NO:2 wherein T is replaced by U, or
• the sequence complementary to the RNA form of said Staphylococcus aureus sequence homologous to SEQ ID NO:
• 1 wherein T is replaced by U,
• the sequence complementary to the RNA form of said Staphylococcus aureus sequence homologous to SEQ ID NO:2 wherein T is replaced by U,
• (ii) detecting the hybrids formed as a result of (i), and
• (iii) detecting or identifying Staphylococcus aureus in the sample based on the presence of the hybrids detected in (ii)
• wherein the method comprises the step of amplifying the 16S-23S rRNA spacer region, or a fragment comprising the target sequence, or the target sequence in a PCR amplification preceding step (i),
• wherein the PCR amplification comprises the use of the primers of SEQ ID NO:68 and SEQ ID NO:58.