Difficile specific oligonucleotides

Claim 2

An oligonucleotide for use in detecting DNA or RNA from microorganisms of the species Clostridium difficile, said oligonucleotide being selected from the group consisting of the sequences: ______________________________________ i) A 5' ATAGGGGTGT TTAAAGGACC nucleotides TAAAGGATTT GAATATTTTG 1-81 of Seq. CACCTGCTAA TACTTATAAT I.D. No. 1. AATAACATAG AAGGTCAGGC T 3' ii) B 5' ATAGTTTATC AAAGTAAATT nucleotides CTTAACTTTG AATGGTAAAA 82-144 of AATATTACTT TGATAATAAC Seq. I.D. TCA 3' No. 1. iii) C 5' GAAGCAGCTA CTGGATGGCA nucleotides AACTATTGAT GGTAAAAAAT 208-270 of ATTACTTTAA TACTAACACT Seq. I.D. GCT 3' No. 1. iv) D 5' ATAGCTTCAA CTGGTTATAC nucleotides AATTATTAAT GGTAAACATT 334-396 of TTTATTTTAA TACTGATGGT Seq. I.D. ATT 3' No. 1. ______________________________________ v) DNA sequences consisting of sequences that are fully complementary to sequence A, B, C or D; vi) DNA sequences that are partial subsequences of the DNA sequence A, B, C or D, such partial subsequences having a minimum of 15 consecutive nucleotide bases; vii) DNA sequences that are partial subsequences of the DNA sequences that are fully complementary to sequence A, B, C or D, such partial subsequences having a minimum of 15 consecutive nucleotide bases; viii) RNA sequences having a sequence as defined in sequence A, B, C or D, but with U replacing T; and ix) RNA sequences having a partial subsequence of sequence A, B, C or D, the partial subsequence having a minimum of 15 consecutive nucleotide bases and with U replacing T in the partial sequence.

Claim 3

An oligonucleotide according to claim 1, wherein said oligonucleotide is from 15 to 30 nucleotide bases in length.

Claim 4

An oligonucleotide for use in detecting DNA or RNA from microorganisms of the species Clostridium difficile, said oligonucleotide being selected from the group consisting of: ______________________________________ i) A 5' ATAGGGGTGT TTAAAGGACC nucleo- TAAAGGATTT GAATATTTTG tides 1-81 CACCTGCTAA TACTTATAAT of Seq. AATAACATAG AAGGTCAGGC T 3' I.D. No. 1 ii) C 5' GAAGCAGCTA CTGGATGGCA nucleo- AACTATTGAT GGTAAAAAAT tides ATTACTTTAA TACTAACACT GCT 3' 208-270 of Seq. I.D. No. 1 ______________________________________ iii) DNA sequences consisting of sequences that are fully complementary to sequence A and C; iv) DNA sequences that are partial subsequences of the DNA sequence A or C, such partial subsequences having a minimum of 15 consecutive nucleotide bases; v) DNA sequences that are partial subsequences of the DNA sequences that are fully complementary to sequence A or C, such partial subsequences having a minimum of 15 consecutive nucleotide bases; vi) RNA sequences having a sequence as defined in sequence A or C but with U replacing T; and vii) RNA sequences having a partial subsequences of A or C, the partial subsequence having a minimum of 15 consecutive nucleotide bases and with U replacing T in the partial sequence.

Claim 5

An oligonucleotide for use in detecting DNA or RNA from microorganisms of the species Clostridium difficile, said oligonucleotide being selected from the group consisting of: ______________________________________ i) A 5' ATAGGGGTGT TTAAAGGACC nucleo- TAAAGGATTT GAATATTTTG tides 1-81 CACCTGCTAA TACTTATAAT of Seq. AATAACATAG AAGGTCAGGC T 3' I.D. No. 1 ii) C 5' GAAGCAGCTA CTGGATGGCA nucleo- AACTATTGAT GGTAAAAAAT tides ATTACTTTAA TACTAACACT GCT 3' 208-270 of Seq. I.D. No. 1 ______________________________________ iii) DNA sequences consisting of sequences that are fully complementary to sequence A and C; iv) DNA sequences that are partial subsequences of the DNA sequence A or C, such partial subsequences having a minimum of 15 consecutive nucleotide bases; v) DNA sequences that are partial subsequences of the DNA sequences that are fully complementary to sequence A or C, such partial subsequences having a minimum of 15 consecutive nucleotide bases; vi) RNA sequences having a sequence as defined in sequence A or C but with U replacing T; and vii) RNA sequences having a partial subsequences of A or C, the partial subsequence having a minimum of 15 consecutive nucleotide bases and with U replacing T in the partial sequence.

Claim 6

An oligonucleotide according to claim 3, wherein said oligonucleotide is from 15 to 30 nucleotide bases in length.

Claim 7

The oligonucleotide 5' GAAGCAGCTA CTGGATGGCA 3' nucleotides 208-226 of Seq. I.D. No.1.

Claim 8

The oligonucleotide 5' GAAGCAGCTA CTGGATGGCA 3' nucleotides 208-226 of Seq. I.D. No.1.

Claim 9

The oligonucleotide 5' AGCAGTGTTA GTATTAAAGT 3' complementary to nucleotides 251-270 of Seq. I.D. No.1.

Claim 10

The oligonucleotide 5' AGCAGTGTTA GTATTAAAGT 3' complementary to nucleotides 251-270 of Seq. I.D. No.1.

Claim 11

The oligonucleotide 5' ATAGGGGTGT TTAAAGGACC T 3' nucleotides 1.21 of Seq. I.D. No.1.

Claim 12

The oligonucleotide 5' ATAGGGGTGT TTAAAGGACC T 3' nucleotides 1.21 of Seq. I.D. No.1.

Claim 13

The oligonucleotide 5' AGCCTGACCT TCTATGTTAT 3' complementary to nucleotides 62-81 of Seq. I.D. No.1.

Claim 14

The oligonucleotide 5' AGCCTGACCT TCTATGTTAT 3' complementary to nucleotides 62-81 of Seq. I.D. No.1.

Claim 15

An oligonucleotide according to claim 1, wherein said oligonucleotide is a DNA sequence.

Claim 16

An oligonucleotide according to claim 3, wherein said oligonucleotide is a DNA sequence.

Claim 17

An oligonucleotide probe for use in detecting DNA or RNA from microorganisms of the species Clostridium difficile, said probe consisting of an oligonucleotide sequence labelled with a label permitting detection of the probe when hybridized to a complementary sequence of C. difficile DNA or mRNA, said oligonucleotide sequence being selected from the group consisting of ______________________________________ i) A 5' ATAGGGGTGT TTAAAGGACC nucleotides TAAAGGATTT GAATATTTTG 1-81 of Seq. CACCTGCTAA TACTTATAAT I.D. No. 1. AATAACATAG AAGGTCAGGC T 3' ii) B 5' ATAGTTTATC AAAGTAAATT nucleotides CTTAACTTTG AATGGTAAAA 82-144 of AATATTACTT TGATAATAAC Seq. I.D. TCA 3' No. 1. iii) C 5' GAAGCAGCTA CTGGATGGCA nucleotides AACTATTGAT GGTAAAAAAT 208-270 of ATTACTTTAA TACTAACACT Seq. I.D. GCT 3' No. 1. iv) D 5' ATAGCTTCAA CTGGTTATAC nucleotides AATTATTAAT GGTAAACATT 334-396 of TTTATTTTAA TACTGATGGT Seq. I.D. ATT 3' No. 1. ______________________________________ v) DNA sequences consisting of sequences that are fully complementary to sequence A, B, C or D; vi) DNA sequences that are partial subsequences of the DNA sequence A, B, C or D, such partial subsequences having a minimum of 15 consecutive nucleotide bases; vii) DNA sequences that are partial subsequences of the DNA sequence that are fully complementary to sequence A, B, C or D, such partial subsequences having a minimum of 15 consecutive nucleotide bases; viii) RNA sequences having a sequence as defined in sequence A, B, C, or D but with U replacing T; and ix) RNA sequences having a partial subsequence A, B, C or D, the partial subsequence having a minimum of 15 consecutive nucleotide bases and with U replacing T in the partial sequence.

Claim 18

An oligonucleotide probe for use in detecting DNA or RNA from microorganisms of the species Clostridium difficile, said probe consisting of an oligonucleotide sequence labelled with a label permitting detection of the probe when hybridized to a complementary sequence of C. difficile DNA or mRNA, said oligonucleotide sequence being selected from the group consisting of ______________________________________ i) A 5' ATAGGGGTGT TTAAAGGACC nucleotides TAAAGGATTT GAATATTTTG 1-81 of Seq. CACCTGCTAA TACTTATAAT I.D. No. 1. AATAACATAG AAGGTCAGGC T 3' ii) B 5' ATAGTTTATC AAAGTAAATT nucleotides CTTAACTTTG AATGGTAAAA 82-144 of AATATTACTT TGATAATAAC Seq. I.D. TCA 3' No. 1. iii) C 5' GAAGCAGCTA CTGGATGGCA nucleotides AACTATTGAT GGTAAAAAAT 208-270 of ATTACTTTAA TACTAACACT Seq. I.D. GCT 3' No. 1. iv) D 5' ATAGCTTCAA CTGGTTATAC nucleotides AATTATTAAT GGTAAACATT 334-396 of TTTATTTTAA TACTGATGGT Seq. I.D. ATT 3' No. 1. ______________________________________ v) DNA sequences consisting of sequences that are fully complementary to sequence A, B, C or D; vi) DNA sequences that are partial subsequences of the DNA sequence A, B, C or D, such partial subsequences having a minimum of 15 consecutive nucleotide bases; vii) DNA sequences that are partial subsequences of the DNA sequence that are fully complementary to sequence A, B, C or D, such partial subsequences having a minimum of 15 consecutive nucleotide bases; viii) RNA sequences having a sequence as defined in sequence A, B, C, or D but with U replacing T; and ix) RNA sequences having a partial subsequence A, B, C or D, the partial subsequence having a minimum of 15 consecutive nucleotide bases and with U replacing T in the partial sequence.

Claim 19

An oligonucleotide probe according to claim 11, wherein said probe is from 15 to 30 nucleotide bases in length.

Claim 20

An oligonucleotide probe according to claim 11, wherein said oligonucleotide sequence is a DNA sequence.

Claim 21

An oligonucleotide probe according to claim 11 wherein the label attached to the oligonucleotide sequence is selected from the group consisting of radioactive labels, fluorescent labels and enzyme labels.

Claim 22

An oligonucleotide probe according to claim 11, wherein said oligonucleotide sequence is immobilized onto a solid support.

Claim 23

An oligonucleotide probe for use in detecting DNA or RNA from microorganisms of the species Clostridium difficile, said probe consisting of an oligonucleotide sequence labelled with a label permitting detection of the probe when hybridized to a complementary sequence of C. difficile DNA or mRNA, said oligonucleotide sequence being selected from the group consisting of ______________________________________ i) A 5' ATAGGGGTGT TTAAAGGACC nucleo- TAAAGGATTT GAATATTTTG tides 1-81 CACCTGCTAA TACTTATAAT of Seq. AATAACATAG AAGGTCAGGC T 3' I.D. No. 1 ii) C 5' GAAGCAGCTA CTGGATGGCA nucleo- AACTATTGAT GGTAAAAAAT tides ATTACTTTAA TACTAACACT GCT 3' 208-270 of Seq. I.D. No. 1 ______________________________________ iii) DNA sequences consisting of sequences that are fully complementary to sequence A or C; iv) DNA sequences that are partial subsequences of the DNA sequence A or C, such partial subsequences having a minimum of 15 consecutive nucleotide bases; v) DNA sequences that are partial subsequences of the DNA sequences that are fully complementary to sequence A or C, such partial subsequences having a minimum of 15 consecutive nucleotide bases; vi) RNA sequences having a sequence as defined in sequence A or C but with U replacing T; and vii) RNA sequences having a partial subsequences of A or C, the partial subsequence having a minimum of 15 consecutive nucleotide bases and with U replacing T in the partial sequence.

Claim 24

An oligonucleotide probe for use in detecting DNA or RNA from microorganisms of the species Clostridium difficile, said probe consisting of an oligonucleotide sequence labelled with a label permitting detection of the probe when hybridized to a complementary sequence of C. difficile DNA or mRNA, said oligonucleotide sequence being selected from the group consisting of ______________________________________ i) A 5' ATAGGGGTGT TTAAAGGACC nucleo- TAAAGGATTT GAATATTTTG tides 1-81 CACCTGCTAA TACTTATAAT of Seq. AATAACATAG AAGGTCAGGC T 3' I.D. No. 1 ii) C 5' GAAGCAGCTA CTGGATGGCA nucleo- AACTATTGAT GGTAAAAAAT tides ATTACTTTAA TACTAACACT GCT 3' 208-270 of Seq. I.D. No. 1 ______________________________________ iii) DNA sequences consisting of sequences that are fully complementary to sequence A or C; iv) DNA sequences that are partial subsequences of the DNA sequence A or C, such partial subsequences having a minimum of 15 consecutive nucleotide bases; v) DNA sequences that are partial subsequences of the DNA sequences that are fully complementary to sequence A or C, such partial subsequences having a minimum of 15 consecutive nucleotide bases; vi) RNA sequences having a sequence as defined in sequence A or C but with U replacing T; and vii) RNA sequences having a partial subsequences of A or C, the partial subsequence having a minimum of 15 consecutive nucleotide bases and with U replacing T in the partial sequence.

Claim 25

An oligonucleotide probe according to claim 16, wherein said probe is from 15 to 30 nucleotide bases in length.

Claim 26

An oligonucleotide probe according to claim 17, wherein said oligonucleotide sequence is a DNA sequence.

Claim 27

An oligonucleotide probe according to claim 17 wherein the label attached to the oligonucleotide sequence is selected from the group consisting of radioactive labels, fluorescent labels and enzyme labels.

Claim 28

An oligonucleotide probe according to claim 17 wherein the oligonucleotide sequence is immobilized onto a solid support.

Claim 29

A method of detecting the presence of C. difficile DNA or mRNA in a sample which comprises treating the sample with an oligonucleotide probe as claimed in claim 28 and under conditions such that the probe can hybridize to the genetic DNA or mRNA so that a hybridication product is formed, and detecting the presence of said DNA or mRNA in the sample by detecting the presence of the hybridized labelled probe.

Claim 30

A method according to claim 21 wherein the oligonucleotide probe is from 15 to 30 nucleotide bases in length.

Claim 31

A method according to claim 22 wherein the oligonucleotide probe is a DNA probe.

Claim 32

A method of amplifying C. difficile DNA in a sample which comprises subjecting said DNA to a polymerase chain reaction using as primers a pair of oligonucleotides as claimed in claim 1, the oligonucleotide sequences used as primers being DNA sequences.

Claim 33

A method of amplifying C. difficile DNA in a sample which comprises subjecting said DNA to a polymerase chain reaction using as primers a pair of oligonucleotides as claimed in claim 3, the oligonucleotide sequences used as primers being DNA sequences.

Claim 34

A method according to claim 25 wherein the primers used are the oligonucleotides: 5' GAAGCAGCTA CTGGATGGCA 3' nucleotides 208-226 of Seq. I.D. No.1, 5' AGCAGTGTTA GTATTAAAGT 3' complementary of nucleotides 251-270 to Seq. I.D. No.1.

Claim 35

A method according to claim 25 wherein the primers used are the oligonucleotides: 5' ATAGGGGTGT TTAAAGGACC T 3' nucleotides 1-27 of Seq. I.D. No.1, 5' AGCCTGACCT TCTATGTTAT 3' complementary of nucleotides 62-81 to Seq. I.D. No.1.

Claim 36

A method according to claim 21 wherein the sample is a clinical sample.

Claim 37

A method according to claim 28 wherein the sample is a faecal sample.