Development of a real-time PCR assay for detection of pneumococcal DNA and diagnosis of pneumococccal disease
원문보기
IPC분류정보
국가/구분
United States(US) Patent
등록
국제특허분류(IPC7판)
C07H-021/04
C07H-021/00
출원번호
US-0089938
(2005-03-25)
등록번호
US-7476733
(2009-01-13)
발명자
/ 주소
Carvalho,Maria da Gloria
Sampson,Jacquelyn S.
Ades,Edwin W.
Carlone,George
McCaustland,Karen
출원인 / 주소
The United States of America as represented by the Department of Health and Human Services
대리인 / 주소
Klarquist Sparkman, LLP.
인용정보
피인용 횟수 :
5인용 특허 :
242
초록
Disclosed are compositions and methods for detecting a specific sequence of the psaA gene using real-time PCR for diagnosis of Pneumococcal Disease.
대표청구항▼
What is claimed is: 1. An oligonucleotide primer for the amplification of a Streptococcus pneumoniae psa nucleic acid, wherein the oligonucleotide primer is 15 to 30 nucleotides in length and hybridizes, under conditions suitable for a polymerase chain reaction, with the nucleic acid sequence accor
What is claimed is: 1. An oligonucleotide primer for the amplification of a Streptococcus pneumoniae psa nucleic acid, wherein the oligonucleotide primer is 15 to 30 nucleotides in length and hybridizes, under conditions suitable for a polymerase chain reaction, with the nucleic acid sequence according to SEQ ID NO: 4, wherein the oligonucleotide primer comprises at least 15 consecutive nucleotides of the nucleic acid sequence according to SEQ ID NO: 1, and wherein the oligonucleotide primer is capable of directing the amplification of the Streptococcus pneumoniae psa nucleic acid. 2. A composition comprising the oligonucleotide primer of claim 1. 3. A kit comprising reagents for real-time PCR-type amplification reaction for detecting Streptococcus pneumoniae, comprising sense primers, antisense primers and a nondegenerate probe, wherein the sense primer is the oligonucleotide primer of claim 1. 4. An oligonucleotide primer for the amplification of a Streptococcus pneumoniae psaA nucleic acid, wherein the oligonucleotide primer is 15 to 30 nucleotides in length and hybridizes, under conditions suitable for a polymerase chain reaction, with the nucleic acid sequence according to SEQ ID NO: 4, wherein the oligonucleotide primer comprises at least 15 consecutive nucleotides of the nucleic acid sequence according to SEQ ID NO: 2, and wherein the oligonucleotide primer is capable of directing the amplification of the Streptococcus pneumoniae psaA nucleic acid. 5. A composition comprising the oligonucleotide primer of claim 4. 6. A kit comprising reagents for real-time PCR-type amplification reaction for detecting Streptococcus pneumoniae, comprising sense primers, antisense primers and a nondegenerate probe, wherein the antisense primer is the oligonucleotide primer of claim 4. 7. An oligonucleotide probe for the detection of a Streptococcus pneumoniae psa nucleic acid, wherein the oligonucleotide probe is 20 to 35 nucleotides in length and hybridizes, under conditions suitable for a polymerase chain reaction, with the nucleic acid sequence according to SEQ ID NO: 4 or the complement thereof, and wherein the oligonucleotide probe comprises at least 20 consecutive nucleotides of the nucleic acid sequence according to SEQ ID NO: 3 or SEQ ID NO: 7. 8. A composition comprising the oligonucleotide probe of claim 7. 9. The oligonucleotide probe of claim 7, wherein the oligonucleotide probe comprises the nucleotide sequence 5'-X-CTAGCACATGC "T"ACAAGAATGATTGCAGAAAGAAA-Y-3' (SEQ ID NO: 3), wherein X is a fluorophore, wherein Y is a phosphate group or phosphate groups, and wherein "T" is a thymine with a dark quencher or acceptor dye linked to it. 10. A kit comprising reagents for real-time PCR-type amplification reaction for detecting Streptococcus pneumoniae, comprising sense primers, and antisense primers and a nondegenerate probe, wherein the nondegenerate probe is the oligonucleotide probe of claim 7. 11. A kit comprising reagents for real-time PCR-type amplification reaction for detecting Streptococcus pneumoniae, comprising a sense primer, an antisense primer and a nondegenerate probe, wherein the sense primer is a nucleic acid molecule consisting of the nucleic acid sequence according to SEQ ID NO: 1; the antisense primer is a nucleic acid molecule consisting of the nucleic acid sequence according to SEQ ID NO: 2; and the non-degenerate probe is a nucleic acid molecule consisting of the nucleic acid sequence according to 5'-X-CTAGCACATGC"T"ACAAGAATGATTGCAGAAAGAAA-Y-3' (SEQ ID NO: 3), wherein X is a fluorophore, wherein Y is a phosphate group or phosphate groups, and wherein "T" is a thymine with a dark quencher or acceptor dye linked to it. 12. The oligonucleotide primer of claim 1, wherein the oligonucleotide primer is 25-30 nucleotides is length. 13. The oligonucleotide primer of claim 1, wherein the oligonucleotide primer hybridizes, under conditions suitable for a polymerase chain reaction, with the nucleic acid according to SEQ ID NO: 5. 14. The oligonucleotide primer of claim 1, wherein the oligonucleotide primer consists of a nucleic acid molecule with the nucleic acid sequence according to SEQ ID NO: 1. 15. The oligonucleotide primer of claim 4, wherein the oligonucleotide primer is 25-30 nucleotides is length. 16. The oligonucleotide primer of claim 4, wherein the oligonucleotide primer hybridizes, under conditions suitable for a polymerase chain reaction, with the nucleic acid according to SEQ ID NO: 6. 17. The oligonucleotide primer of claim 4, wherein the oligonucleotide primer consists of the nucleic acid sequence according to SEQ ID NO: 2. 18. The oligonucleotide primer of claim 7, wherein the oligonucleotide probe is 27-35 nucleotides is length. 19. The oligonucleotide probe of claim 7, wherein the oligonucleotide probe consists of the nucleotide sequence 5'-X-CTAGCACATGC "T"ACAAGAATGATTGCAGAAAGAAA-Y-3' (SEQ ID NO: 3), wherein X is a fluorophore, wherein Y is a phosphate group or phosphate groups, and wherein "T" is a thymine with a dark quencher or acceptor dye linked to it.
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