Polynucleotide amplification mycoplasma assay, primers, and kits therefore
AssigneeStratagene
InventorsDouglas T. McKenzie, Joseph A. Sorge
Life of this patent
Estimating the term…
Abstract
The invention described herein consists of methods and materials for the detection of mycoplasma infections. Mycoplasma infections may be detected in cell cultures and in animals. The subject methods use polynucleotide primer pairs that are capable of hybridizing to mycoplasma tRNA genes so as to provide for the generation of a distinctive set of amplification products when the primers are used in a cyclic amplification synthesis reaction, such as PCR (polymerase chain reaction). In addition to detecting mycoplasma infections, the subject methods may be used to identify the particular species of mycoplasma causing the infection. The subject invention also provides for primers and kits for performing the subject methods.
Claims8
A method of detecting mycoplasma in a sample for analysis, said method comprising the steps,
- isolating polynucleotides from said sample for analysis, whereby a polynucleotide sample is produced
- adding a pair of polynucleotide primers which hybridize to a mycoplasma tRNA gene, to said polynucleotide sample,
- amplifying at least a portion of at least one tRNA gene cluster by a cyclic amplification reaction employing said primers, whereby at least one amplification product is produced, and detecting said amplified product, wherein said detection of amplified probe is indicative of the presence of mycoplasma,
- wherein said pair of primers is selected from the group consisting of:
- ______________________________________ (a) Primer 772 5'CGAAGGGTCGCAGGTTCAAA3' (SEQ ID No:
- 1), and Primer 773 5'CTCAGTCGGTAGAGCAGCAGA3' (SEQ ID No:
- 2), (b) Primer 772 5'CGAAGGGTCGCAGGTTCAAA3' (SEQ ID No:
- 1), and Primer 965 5'AGTCCGGTGCTCTAACCAACTGAG3' (SEQ ID No:
- 5), (c) Primer 964 5'AGGTCGCGGGTTCGAATCC3' (SEQ ID No:
- 4), and Primer 773 5'CTCAGTCGGTAGAGCAGCAGA3' (SEQ ID No:
- 2), (d) Primer 964 5'AGGTCGCGGGTTCGAATCC3' (SEQ ID No:
- 4), and Primer 774, 5'CTCAGTCGGTTAGAGCAGCAG3' (SEQ ID No:
- 3), (e) Primer 415 5'CGGAGGYCGCAGGTTCGAG3' (SEQ ID No:
- 6), and Primer 965 5'AGTCCGGTGCTCTAACCAACTGAG3' (SEQ ID No:
- 5), (f) Primer 415 5'CGGAGGYCGCAGGTTCGAG3' (SEQ ID No:
- 6), and Primer 773 5'CTCAGTCGGTAGAGCAGCAGA3' (SEQ ID No:
- 2), (g) Primer 415 5'CGGAGGYCGCAGGTTCGAG3' (SEQ ID No:
- 6), and Primer 774 5'CTCAGTCGGTTAGAGCAGCAG3' (SEQ ID No:
- 3), and (h) Primer 773 5'CTCAGTCGGTAGAGCAGCAGA3' (SEQ ID No:
- 2), and Primer 772 5'CGAAGGGTCGCAGGTTCAAA3' (SEQ ID No:
- 1). ______________________________________
A polynucleotide primer wherein the primer consists of the nucleotide sequence selected from the group consisting of:
- ______________________________________ Primer 772 5'CGAAGGGTCGCAGGTTCAAA3' (SEQ ID No:
- 1), Primer 773 5'CTCAGTCGGTAGAGCAGCAGA3' (SEQ ID No:
- 2), Primer 774, 5'CTCAGTCGGTTAGAGCAGCAG3' (SEQ ID No:
- 3), and Primer 415 5'CGGAGGYCGCAGGTTCGAG3' (SEQ ID No:
- 6). ______________________________________
A pair of polynucleotide primers, wherein said primers is selected from the group consisting of:
- ______________________________________ (a) Primer 772 5'CGAAGGGTCGCAGGTTCAAA3' (SEQ ID No:
- 1), and Primer 773 5'CTCAGTCGGTAGAGCAGCAGA3' (SEQ ID No:
- 2), (b) Primer 772 5'CGAAGGGTCGCAGGTTCAAA3' (SEQ ID No:
- 1), and Primer 965 5'AGTCCGGTGCTCTAACCAACTGAG3' (SEQ ID No:
- 5), (c) Primer 964 5'AGGTCGCGGGTTCGAATCC3' (SEQ ID No:
- 4), and Primer 773 5'CTCAGTCGGTAGAGCAGCAGA3' (SEQ ID No:
- 2), (d) Primer 964 5'AGGTCGCGGGTTCGAATCC3' (SEQ ID No:
- 4), and Primer 774, 5'CTCAGTCGGTTAGAGCAGCAG3' (SEQ ID No:
- 3), (e) Primer 415 5'CGGAGGYCGCAGGTTCGAG3' (SEQ ID No:
- 6), and Primer 965 5'AGTCCGGTGCTCTAACCAACTGAG3' (SEQ ID No:
- 5), (f) Primer 415 5'CGGAGGYCGCAGGTTCGAG3' (SEQ ID No:
- 6), and Primer 773 5'CTCAGTCGGTAGAGCAGCAGA3' (SEQ ID No:
- 2), (g) Primer 415 5'CGGAGGYCGCAGGTTCGAG3' (SEQ ID No:
- 6), and Primer 774 5'CTCAGTCGGTTAGAGCAGCAG3' (SEQ ID No:
- 3), and (h) Primer 773 5'CTCAGTCGGTAGAGCAGCAGA3' (SEQ ID No:
- 2), and Primer 772 5'CGAAGGGTCGCAGGTTCAAA3' (SEQ ID No:
- 1). ______________________________________
Dependencies are inferred from the claim text.