PNA probes, probe sets, methods and kits pertaining to the universal detection of bacteria and eucarya

A PNA probe set wherein at least one PNA probe of the set has a probing nucleobase sequence of which at least a portion is at least ninety percent homologous to the nucleobase sequences, or their complements, selected from the group consisting of:

• CTG-CCT-CCC-GTA-GGA (SEQ ID NO:1);
• TAC-CAG-GGT-ATC-TAA-T (SEQ ID NO:2);
• CAC-GAG-CTG-ACG-ACA (SEQ ID NO:3);
• CCG-ACA-AGG-AAT-TTC (SEQ ID NO:4);
• ACC-AGA-CTT-GCC-CTC-C (SEQ ID NO:5);
• GGG-CAT-CAC-AGA-CCT-G (SEQ ID NO:6);
• TAG-AAA-GGG-CAG-GGA (SEQ ID NO:7) and TAC-AAA-GGG-CAG-GGA (SEQ ID NO:8).

A method for detecting, identifying or quantitating bacteria and/or eucarya in a sample, said method comprising:

• a) contacting the sample with one or more PNA probes wherein at least one PNA probe has a probing nucleobase sequence of which at least a portion is at least ninety percent homologous to the nucleobase sequences, or their complements, selected from the group consisting of:
• CTG-CCT-CCC-GTA-GGA (SEQ ID NO:1);
• TAC-CAG-GGT-ATC-TAA-T (SEQ ID NO:2);
• CAC-GAG-CTG-ACG-ACA (SEQ ID NO:3);
• CCG-ACA-AGG-AAT-TTC (SEQ ID NO:4);
• ACC-AGA-CTT-GCC-CTC-C (SEQ ID NO:5);
• GGG-CAT-CAC-AGA-CCT-G (SEQ ID NO:6);
• TAG-AAA-GGG-CAG-GGA (SEQ ID NO:7) and TAC-AAA-GGG-CAG-GGA (SEQ ID NO:8);
• and
• b) detecting, identify or quantitating hybridization of the probing nucleobase sequence of PNA probes to the target sequences of bacteria or eucarya, and correlating the result with the presence, absence or number of the bacteria or eucarya in the sample.

A kit suitable for performing an assay which detects, identifies or enumerates bacteria or eucarya in a sample, said kit comprising:

• a) one or more PNA probes, wherein the PNA probes each having a probing nucleobase sequence of which at least a portion is at least ninety percent homologous to the nucleobase sequences, or their complements, selected from the group consisting of:
• CTG-CCT-CCC-GTA-GGA (SEQ ID NO:1);
• TAC-CAG-GGT-ATC-TAA-T (SEQ ID NO:2);
• CAC-GAG-CTG-ACG-ACA (SEQ ID NO:3);
• CCG-ACA-AGG-AAT-TTC (SEQ ID NO:4);
• ACC-AGA-CTT-GCC-CTC-C (SEQ ID NO:5);
• GGG-CAT-CAC-AGA-CCT-G (SEQ ID NO:6);
• TAG-AAA-GGG-CAG-GGA (SEQ ID NO:7) and TAC-AAA-GGG-CAG-GGA (SEQ ID NO:8);
• and
• b) other reagents or compositions necessary to perform the assay.

A multiplex PNA-ISH assay for the universal detection, identification or quantitation of bacteria and eucarya;

• the assay comprising:
• a) contacting a sample with two or more independently detectable PNA probes, wherein the two or more independently detectable PNA probes each comprise a probing nucleobase sequence which hybridizes to a target sequence;
• such that least one detectable PNA probe hybridizes to a target sequence in bacteria and at least one other independently detectable PNA probe hybridizes to a target sequence in eucarya;
• and
• b) detecting, identify or quantitating hybridization of the probing nucleobase sequence of the PNA probes to the target sequence of the different organisms and correlating the result for each of the two or more independently detectable PNA probes with the presence, absence or number of bacteria and eucarya in the sample, wherein at least one detectable PNA probe used in the assay has a probing nucleobase sequence of which at least a portion is at least ninety percent homologous to the nucleobase sequences, or their complements, selected from the group consisting of:
• CTG-CCT-CCC-GTA-GGA (SEQ ID NO:1);
• TAC-CAG-GGT-ATC-TAA-T (SEQ ID NO:2);
• CAC-GAG-CTG-ACG-ACA (SEQ ID NO:3) and CCG-ACA-AGG-AAT-TTC (SEQ ID NO:4);
• and at least one other independently detectable PNA probe has a probing nucleobase sequence of which at least a portion is at least ninety percent homologous to the nucleobase sequences, or their complements, selected from the group consisting of:
• ACC-AGA-CTT-GCC-CTC-C (SEQ ID NO:5);
• GGG-CAT-CAC-AGA-CCT-G (SEQ ID NO:6);
• TAG-AAA-GGG-CAG-GGA (SEQ ID NO:7);
• and TAC-AAA-GGG-CAG-GGA (SEQ ID NO:8).