Methods and compositions for detecting bacterial contamination
AssigneeDch Molecular Diagnostics, Inc.
InventorsJames Jian Quan Want, Xiangmin Cui, HiuNam Chan
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Abstract
Compositions, reactions mixtures, kits, and systems for detecting bacterial contamination are provided, as well as methods of using the same.
Claims39
A method of detecting bacterial contamination of a platelet sample by any of at least five bacterial genera, comprising:
- (a) subjecting the sample or a portion thereof to a nucleic acid amplification reaction under conditions to yield a detectable amount of an amplicon of no more than about 800 bases in a single reaction mixture, said reaction mixture comprising a single primer pair and a single detectable probe, wherein the primer pair flanks the amplicon, the amplicon comprises a conserved sequence to which the detectable probe hybridizes, and the conserved sequence is identical among the at eight-five bacterial genera;
- and
- (b) detecting hybridization to the amplicon by the probe, wherein the hybridization by the probe yields a detectable signal indicative of bacterial contamination of the platelet sample by any of the at least five bacterial genera.
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A composition for amplification and detection of a portion of a 16S rRNA polynucleotide, said portion being less than about 800 nucleotides in length, comprising:
- a first primer comprising at least 10 contiguous nucleotides of the sequence as set forth in SEQ ID NO:
- 9;
- a second primer comprising at least 10 contiguous nucleotides of the sequence as set forth in SEQ ID NO:
- 10;
- and,
- a probe comprising at least 10 contiguous nucleotides of the sequence as set forth in SEQ ID NO:
- 16, or the complement thereof.
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A reaction mixture for amplification and detection of a portion of a 16S rRNA polynucleotide, said portion being less than about 800 nucleotides in length, comprising
- sample nucleic acid;
- primers that, in an amplification reaction with a target 16S rRNA polynucleotide, amplify an amplicon of at least 50 nucleotides in length, the amplicon having 90% sequence identity with any of SEQ ID NO:
- 1-3 when optimally aligned;
- a probe that specifically hybridizes to either strand of the amplicon;
- and
- a polymerase;
- wherein the reaction mixture is in a reaction site.
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Dependencies are inferred from the claim text.