초록 ▼

The present invention relates to DNA-based methods for universal bacterial detection, for specific detection of the common bacterial pathogens Escherichia coli, Klebsiella pneumoniae, Pseudomonas aeruginosa, Proteus mirabilis, Streptococcus pneumoniae, Staphylococcus aureus, Staphylococcus epidermid

The present invention relates to DNA-based methods for universal bacterial detection, for specific detection of the common bacterial pathogens Escherichia coli, Klebsiella pneumoniae, Pseudomonas aeruginosa, Proteus mirabilis, Streptococcus pneumoniae, Staphylococcus aureus, Staphylococcus epidermidis, Enterococcus faecalis, Staphylococcus saprophyticus, Streptococcus pyogenes, Haemophilus influenzae and Moraxella catarrhalis as well as for specific detection of commonly encountered and clinically relevant bacterial antibiotic resistance genes directly from clinical specimens or, alternatively, from a bacterial colony. The above bacterial species can account for as much as 80% of bacterial pathogens isolated in routine microbiology laboratories.

대표청구항 ▼

[ What is claimed is:] [1.] A method using probes or amplification primers or both which are specific, ubiquitous and sensitive for determining the presence or amount of nucleic acids:from a bacterial antibiotic resistance gene selected from the group consisting of bla.sub.tem, bla.sub.rob, bla.sub.

[ What is claimed is:] [1.] A method using probes or amplification primers or both which are specific, ubiquitous and sensitive for determining the presence or amount of nucleic acids:from a bacterial antibiotic resistance gene selected from the group consisting of bla.sub.tem, bla.sub.rob, bla.sub.shv, aadB, aacC1, aacC2, aacC3, aacA4, mecA, vanA, vanH, vanX, satA, aacA-aphH, vat, vga, msrA sul, and int, andfrom specific bacterial species selected from the group consisting of Escherichia coli, Klebsiella pneumoniae, Pseudomonas aeruginosa, Proteus mirabilis, Streptococcus pneumoniae, Staphylococcus aureus, Staplylococcus epidermidis, Enterococcus faecalis, Staphylococcus epidermidis, Enterococcus faecalis, Staphylococcus saprophyticus, Streptococcus pyogenes, Haemophilus influenzae, and Moraxella catarrhalis, in any sample suspected of containing said nucleic acids,wherein each of said nucleic acids comprises a selected target region hybridizable with said probes or primers;said method comprising the steps of contacting said sample with said probes or primers and detecting the presence or amount of hybridized probes or amplified products as an indication of the presence or amount of said specific bacterial species simultaneously with said bacterial antibiotic resistance gene;said probes or primers comprising at least one single stranded nucleic acid which nucleotidic sequence has at least twelve nucleotides in length capable of hybridizing with said bacterial species and with any one of:SEQ ID NO. 3, SEQ ID NO. 4, SEQ ID NO. 5, SEQ ID NO. 6, SEQ ID NO. 7 and a complementary sequence thereof, for determining the presence or amount of Escherichia coli;SEQ ID NO. 8, SEQ ID NO. 9, SEQ ID NO. 10, SEQ ID NO. 11, and a complementary sequence thereof, for determining the presence or amount of Klebsiella pneumoniae;SEQ ID NO. 16, SEQ ID NO. 17, SEQ ID NO. 18, SEQ ID NO. 19, SEQ ID NO. 20 and a complementary sequence thereof, for determining the presence or amount of Pseudomonas aeruginosa;SEQ ID NO. 12, SEQ ID NO. 13, SEQ ID NO. 14, SEQ ID NO. 15 and a complementary sequence thereof, for determining the presence or amount of Proteus mirabilis;SEQ ID NO. 30, SEQ ID NO. 31, SEQ ID NO. 34, SEQ ID NO. 35 and complementary sequence thereof, for determining the presence or amount of Streptococcus pneumoniae;SEQ ID NO. 37 and a complementary sequence thereof, for determining the presence or amount of Staplylococcus aureus;SEQ ID NO. 36 and a complementary sequence thereof, for determining the presence or amount of Staphylococcus epidermidis;SEQ ID NO. 1, SEQ ID NO. 2 and a complementary sequence thereof, for determining the presence or amount of Enterococcus faecalis;SEQ ID NO. 21, SEQ ID NO. 22, SEQ ID NO. 23, SEQ ID NO. 24, and a complementary sequence thereof, for determining the presence or amount of Staphylococcus saprophyticus;SEQ ID NO. 32, SEQ ID NO. 33 and a complementary sequence thereof, for determining the presence or amount of Streptococcus pyogenes;SEQ ID NO. 25, SEQ ID NO. 26, SEQ ID NO. 27 and a complementary sequence thereof, for determining the presence or amount of Haemophilus influenzae; andSEQ ID NO. 28, SEQ ID NO. 29 and a complementary sequence thereof, for determining the presence or amount of Moraxella catarrhalis.