Diagnostic assay for antibiotic tolerance
AssigneeSt. Jude Children's Research Hospital
InventorsRobyn M. Atkinson, Elaine I. Tuomanen
Life of this patent
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Abstract
Allelic variation in the vex2, pep27 and vncS genes of bacteria responsible for tolerance to antibiotics such as penicillin and vancomycin, is taught. Methods for identifying antibiotic tolerant bacteria and subjects infected with such bacteria, particularly antibiotic tolerant Streptococcus pneumoniae, are provided. Test kits and components useful for performing such methods, particularly including oligonucleotide primers, are also provided.
Claims16
A method for determining whether a bacteria is likely to be tolerant to at least one antibiotic comprising
- (a) determining whether said bacteria has a type 4 or R6 allele of the vex2 gene, and
- (b) determining whether said bacteria has a type 4 or R6 allele of the pep27 gene,
- wherein said bacteria is determined to be likely to be tolerant if it has a type 4 allele of the vex2 gene and an R6 allele of the pep27.
A method for determining the vex2 genotype of a selected bacteria comprising determining the identity of the nucleotides located at the positions corresponding to nucleotides 41 and 67 of SEQ ID No. 1 in the genomic DNA of said selected bacteria.
A method for determining the pep27 genotype of a selected bacteria comprising determining the identity of the nucleotides located at the positions corresponding to nucleotides 35 and 46 of SEQ ID No. 3 in the genomic DNA of said selected bacteria.
A test kit suitable for determining the vex2 and pep27 genotype of a bacteria thereby determining whether said bacteria is likely to be tolerant to at least one antibiotic comprising:
- (a) a predetermined amount of a first amplification primer complementary to a region of the vex2 gene 5′ to the location corresponding to nucleotide 41 of SEQ ID No. 1;
- (b) a predetermined amount of a second amplification primer complementary to a region of the vex2 gene 3′ to the location corresponding to nucleotide 67 of SEQ ID No. 1;
- and
- (c) a predetermined amount of a third amplification primer complementary to a region of the pep27 gene 5′ to the location corresponding to nucleotide 35 of SEQ ID No.3;and
- (d) a predetermined amount of a fourth amplification primer complementary to a region of the pep27 gene 3′ to the location corresponding to nucleotide 46 of SEQ ID No. 3;
- (e) other reagents;
- and
- (f) directions for use of said kit,
- wherein said first amplification primer and said second amplification primer can be used to amplify a region of the vex 2 gene that distinguishes the type 4 and R6 alleles and said third amplification primer and said fourth amplification primer can be used to amplify a region of the pep27 gene that distinguishes the type 4 and R6 alleles.
Dependencies are inferred from the claim text.