Method for detecting mycoplasma
First Claim
1. A kit for detecting Mycoplasma in a test sample by a multiplex real time quantitative PCR, whereinthe kit comprises one or more forward primers, two or more reverse primers, one or more probes and a solid support;
- the probe(s) is immobilized onto the solid support;
the probe is a probe for specifically detecting products amplified by use of the forward primer and at least any one of the reverse primers;
the forward primer(s) is an oligonucleotide consisting of a nucleotide sequence, which is selected from the group consisting of nucleotide sequences each consisting of continuous 17 to 30 nucleotides in the nucleotide sequence of SEQ ID No;
1, and which contains a nucleotide sequence (caaggtatccs) at nucleotide positions 14 to 24 in SEQ ID No;
1;
the reverse primers each are an oligonucleotide consisting of a nucleotide sequence, which is selected from the group consisting of nucleotide sequences each consisting of continuous 17 to 26 nucleotides in one or more nucleotide sequences of SEQ ID Nos;
14 and 17 to 20; and
the probe(s) is an oligonucleotide, which consists of a nucleotide sequence selected from the group consisting of nucleotide sequences each consisting of continuous 17 to 26 nucleotides in the nucleotide sequence of SEQ ID No;
33, or which consists of a complementary nucleotide sequence thereto.
1 Assignment
0 Petitions
Accused Products
Abstract
An object of the present invention is to provide a detection method for Mycoplasma by which a greater number of Mycoplasma species can be more quickly and easily detected with high sensitivity and accuracy, a set of a forward primer, a reverse primer and a probe for the detection and a kit containing the set. Mycoplasma in a test sample is detected by a multiplex real time quantitative PCR using one or more forward primers, each of which is an oligonucleotide consisting of a nucleotide sequence, which is selected from the group consisting of nucleotide sequences each consisting of continuous 17 to 30 nucleotides in the nucleotide sequence represented by SEQ ID No: 1, and which contains a nucleotide sequence at nucleotide positions 14 to 24 in SEQ ID No: 1; reverse primers, each of which is an oligonucleotide consisting of a nucleotide sequence, which is selected from the group consisting of nucleotide sequences each consisting of continuous 17 to 26 nucleotides in the one or more nucleotide sequences represented by SEQ ID Nos: 14 and 17 to 20; and a probe(s), which is an oligonucleotide, which consists of a nucleotide sequence selected from the group consisting of nucleotide sequences each consisting of continuous 17 to 26 nucleotides in the nucleotide sequence represented by SEQ ID No: 33, or which consists of a complementary nucleotide sequence thereto.
-
Citations
14 Claims
-
1. A kit for detecting Mycoplasma in a test sample by a multiplex real time quantitative PCR, wherein
the kit comprises one or more forward primers, two or more reverse primers, one or more probes and a solid support; -
the probe(s) is immobilized onto the solid support; the probe is a probe for specifically detecting products amplified by use of the forward primer and at least any one of the reverse primers; the forward primer(s) is an oligonucleotide consisting of a nucleotide sequence, which is selected from the group consisting of nucleotide sequences each consisting of continuous 17 to 30 nucleotides in the nucleotide sequence of SEQ ID No;
1, and which contains a nucleotide sequence (caaggtatccs) at nucleotide positions 14 to 24 in SEQ ID No;
1;
the reverse primers each are an oligonucleotide consisting of a nucleotide sequence, which is selected from the group consisting of nucleotide sequences each consisting of continuous 17 to 26 nucleotides in one or more nucleotide sequences of SEQ ID Nos;
14 and 17 to 20; and
the probe(s) is an oligonucleotide, which consists of a nucleotide sequence selected from the group consisting of nucleotide sequences each consisting of continuous 17 to 26 nucleotides in the nucleotide sequence of SEQ ID No;
33, or which consists of a complementary nucleotide sequence thereto. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10)
-
-
11. A set of a forward primer, a reverse primer and a probe for detecting Mycoplasma in a test sample by a multiplex real time quantitative PCR, wherein the set contains one or more forward primers, two or more reverse primers and one or more probes;
-
the probe(s) is a probe for specifically detecting products amplified by use of the forward primer and at least any one of the reverse primers and the probe is labeled with a fluorescent substance; the forward primer(s) is an oligonucleotide consisting of a nucleotide sequence, which is selected from the group consisting of nucleotide sequences each consisting of continuous 17 to 30 nucleotides in the nucleotide sequence of SEQ ID No;
1, and which contains a nucleotide sequence (caaggtatccs) at nucleotide positions 14 to 24 in SEQ ID No;
1;
the reverse primers each are an oligonucleotide consisting of a nucleotide sequence, which is selected from the group consisting of nucleotide sequences each consisting of continuous 17 to 26 nucleotides in one or more nucleotide sequences of SEQ ID Nos;
14 and 17 to 20; and
the probe(s) is an oligonucleotide, which consists of a nucleotide sequence selected from the group consisting of nucleotide sequences each consisting of continuous 17 to 26 nucleotides in the nucleotide sequence of SEQ ID No;
33, or which consists of a complementary nucleotide sequence thereto.
-
-
12. A method for detecting Mycoplasma in a test sample by a multiplex real time quantitative PCR, comprising
(a) Step a of extracting DNA from the test sample, (b) Step b of performing a multiplex real time quantitative PCR using the DNA extracted in Step a as a template, and one of more forward primers and two or more reverse primers, and (c) Step c of detecting the presence of Mycoplasma in the test sample by detecting a product amplified by the multiplex real time quantitative PCR in Step b by use of one or more probes; -
the probe(s) is a probe for specifically detecting products amplified by use of the forward primer and at least any one of the reverse primers; the forward primer(s) is an oligonucleotide consisting of a nucleotide sequence, which is selected from the group consisting of nucleotide sequences each consisting of continuous 17 to 30 nucleotides in the nucleotide sequence of SEQ ID No;
1, and which contains a nucleotide sequence (caaggtatccs) at nucleotide positions 14 to 24 in SEQ ID No;
1;
the reverse primers each are an oligonucleotide consisting of a nucleotide sequence, which is selected from the group consisting of nucleotide sequences each consisting of continuous 17 to 26 nucleotides in one or more nucleotide sequences of SEQ ID Nos;
14 and 17 to 20; and
the probe(s) is an oligonucleotide, which consists of a nucleotide sequence selected from the group consisting of nucleotide sequences each consisting of continuous 17 to 26 nucleotides in the nucleotide sequence of SEQ ID No;
33, or which consists of a complementary nucleotide sequence thereto. - View Dependent Claims (13, 14)
-
Specification