PRIMER SET SPECIFIC FOR VANCOMYCIN-RESISTANT ENTEROCOCCUS, COMPOSITION COMPRISING THE SAME AND METHOD OF DETECTING VANCOMYCIN-RESISTANT MICROORGANISM ENTEROCOCCUS IN SAMPLE

摘要

A vancomycin-resistant Enterococcus specific primer set, a composition including the primer set, and a method of detecting a vancomycin-resistant Enterococcus sp. microorganism in a sample using the primer set.

主权项

1. A method of detecting at least one Enterococcus genus microorganism expressing a vancomycin resistance gene, the method comprising:
(i) providing at least one Enterococcus sp. specific primer set selected from the group consisting of
a first primer set comprising a polynucleotide which comprises 10 or more contiguous nucleotides selected from a nucleotide sequence of SEQ ID NO: 1 and a polynucleotide that comprises 10 or more contiguous nucleotides selected from a nucleotide sequence of SEQ ID NO: 2;a second primer set comprising a polynucleotide which comprises 10 or more contiguous nucleotides selected from a nucleotide sequence of SEQ ID NO: 4 and a polynucleotide that comprises 10 or more contiguous nucleotides selected from SEQ ID NO: 5;a third primer set comprising a polynucleotide which comprises 10 or more contiguous nucleotides selected from a nucleotide sequence of SEQ ID NO: 7 and a polynucleotide that comprises 10 or more contiguous nucleotides selected from SEQ ID NO: 8; anda fourth primer set comprising a polynucleotide which comprises 10 or more contiguous nucleotides selected from a nucleotide sequence of SEQ ID NO: 10 and a polynucleotide that comprises 10 or more contiguous nucleotides selected from SEQ ID NO: 11; and at least one vancomycin resistance gene specific primer set selected from the group consisting of
a fifth primer set comprising a polynucleotide which comprises 10 or more contiguous nucleotides selected from a nucleotide sequence of SEQ ID NO: 13 and a polynucleotide that comprises 10 or more contiguous nucleotides selected from a nucleotide sequence of SEQ ID NO: 14;a sixth primer set comprising a polynucleotide which comprises 10 or more contiguous nucleotides selected from a nucleotide sequence of SEQ ID NO: 16 and a polynucleotide that comprises 10 or more contiguous nucleotides selected from SEQ ID NO: 17;a seventh primer set comprising a polynucleotide which comprises 10 or more contiguous nucleotides selected from a nucleotide sequence of SEQ ID NO: 19 and a polynucleotide that comprises 10 or more contiguous nucleotides selected from SEQ ID NO: 20; andan eighth primer set comprising a polynucleotide which comprises 10 or more contiguous nucleotides selected from a nucleotide sequence of SEQ ID NO: 22 and a polynucleotide that comprises 10 or more contiguous nucleotides selected from SEQ ID NO: 23; (ii) contacting the primer set specific for at least one Enterococcus and the primer set specific for at least one vancomycin resistance gene with a sample that is suspected to comprise at least one Enterococcus genus microorganism expressing or comprising a vancomycin resistance gene, wherein the primers of the primer set specific for at least one Enterococcus and/or the primers of the primer set specific for at least one vancomycin resistance gene hybridize at least one target nucleic acid selected from the group consisting of an Enterococcus faecalis lactate dehydrogenase gene (ldh), an Enterococcus faecium lactate dehydrogenase gene (ldh), Enterococcus gallinarum D-alanine: D-alanine ligase gene2 (ddl2), an Enterococcus casseliflavus lactate dehydrogenase gene (ldh), a VanA gene, a VanB gene, a VanC1 gene, and a VanC2/3 gene; (iii) elongating the primers hybridized with a target nucleic acid to amplify the target nucleic acid and provide an amplification product; and (iv) identifying the presence of the target nucleic acid in the sample on the basis of the amplification product.