Methods and compositions for isothermal amplification and detection of mycoplasma pneumoniae
AssigneesThe United States of America, as represented by the Secretary, Dept. of Health and Human Services, P3S Corporation.
InventorsJonas M. Winchell, Brianna Petrone, Maureen H. Diaz, Bernard J. Wolff
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Abstract
Disclosed herein are methods and compositions (e.g., oligonucleotide primers) for isothermal amplification and detection of M. pneumoniae nucleic acids in a sample. In some embodiments, the methods include contacting a sample with a set of LAMP primers specific for a M. pneumoniae CARDS toxin-encoding nucleic acid under conditions sufficient to produce an M. pneumoniae nucleic acid amplification product and detecting the resulting M. pneumoniae amplification product. Kits including sets of LAMP primers for detection of M. pneumoniae CARDS toxin nucleic acids are also provided herein.
Claims20
A method of detecting presence of Mycoplasma pneumoniae in a sample, comprising:
- contacting the sample with a set of loop-mediated isothermal amplification (LAMP) primers specific for a M. pneumoniae CARDS toxin-encoding nucleic acid under conditions sufficient for amplification of the M. pneumoniae nucleic acid;
- and
- detecting the M. pneumoniae amplification product, thereby detecting presence of M. pneumoniae in the sample.
A kit comprising a set of loop-mediated isothermal amplification (LAMP) primers specific for a Mycoplasma pneumoniae nucleic acid, comprising a nucleic acid sequence at least 95% identical to of any one of:
- (SEQ ID NO:
- 2)
- CCACCTAGTGATTTGGAAGA;
- (SEQ ID NO:
- 3)
- GGACAAAGAAGATTTTCGAAGTT;
- (SEQ ID NO:
- 4)
- GCTGAACATCAACAAAGAAGGTGCATTGTTGATGAATGTACTACCCA;
- and
- (SEQ ID NO:
- 5)
- ATACCCCACAATTAAGTGGTTGATTCATAGAATATCTGTCCATCTGG.
A buffer comprising 20 mM Tris (pH 8.8), 10 mM KCl, 5.5 mM MgSO4, 10 mM (NH4)2SO4, 0.1% TWEEN-20, 0.8 M betaine, and 1.375 mM MnCl2.
Dependencies are inferred from the claim text.