Compositions and methods for detecting mycobacteria
OwnerAna Maria Zarraga, German Rehren Schweppeinventors
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Abstract
The present invention features compositions and methods that can be used to analyze samples for the presence of mycobacterium from MTC, M. bovis and M. tuberculosis. Moreover, the methods can be used, for example, to differentiate M. bovis from M. tuberculosis and other MTC specimens. The compositions include oligonucleotides that can be used as primers and probes. The primers can be used in multiplex PCR, providing specificity and sensitivity even when used in single run amplifications.
Claims18
An isolated oligonucleotide comprising a sequence represented by SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:7, SEQ ID NO:8, SEQ ID NO:9, SEQ ID NO:10, or SEQ ID NO:11.
A set of oligonucleotides comprising first and second oligonucleotides that function as primers and a third oligonucleotide that functions as a probe, wherein the first, second and third oligonucleotides each have a sequence represented by one of SEQ ID NOs.1-11.
A method of detecting mycobacteria in a sample, the method comprising providing the sample and exposing nucleic acids within the sample to a pair of oligonucleotides that specifically target and facilitate amplification of an IS1081 sequence or an RD4 sequence.
A method comprising
- providing a sample of tissue comprising nodules;
- examining the nodules for granuloma and/or calcification;
- excising small pieces of the tissue surrounding the granuloma or calcification;
- homogenizing the pieces of tissue;
- transferring the homogenized tissue to a microtube containing beads and about 1 ml of DNAzol;
- precipitating the DNA within the tissue;
- drying the precipitate;
- resuspending the precipitate;
- and
- analyzing the precipitated for the presence of RD4 and/or IS1081 target sequences.
Dependencies are inferred from the claim text.