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公开号/公开日
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US2015104789 A1 2015-04-16 [US20150104789] / 2015-04-16
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申请号/申请日
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2013US-14398725 / 2013-05-03
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发明人
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HAAKE DAVID A;HALFORD COLIN;BABBITT JANE T;CHURCHILL BERNARD M;
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申请人
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UNIVERSITY OF CALIFORNIA;UNIVERSITY OF CALIFORNIA LOS ANGELES;US DEPARTMENT OF VETERANS AFFAIRS;
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主分类号
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IPC分类号
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C12Q-001/68
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摘要
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(US20150104789) Described are probes and methods for detecting antibiotic susceptibility of a specimen. The method comprises contacting the specimen with an oligonucleotide probe that specifically hybridizes with a target nucleic acid sequence region of ribosomal RNA. The target sequence is at the junction between a pre-ribosomal RNA tail and mature ribosomal RNA of 23S or 16S rRNA. Performing the method in the presence and absence of an antibiotic permits detection of antibiotic susceptibility.
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机翻摘要
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地址
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代理人
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代理机构
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;
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优先权号
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2012US-61643009 2012-05-04
2013US-14398725 2013-05-03
2013WO-US39574 2013-05-03
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主权利要求
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(US20150104789) 1. A method for determining whether a sample of bacteria is susceptible to an antibiotic agent, the method comprising the steps of: (a) contacting a specimen obtained from the sample with an oligonucleotide probe or pair of probes in the absence of the agent, wherein the probe or pair of probes specifically hybridizes to a target sequence over the full length of the target sequence, wherein the target sequence comprises 25-35 contiguous nucleotides of bacterial ribosomal RNA (rRNA) spanning a splice site between a pre-ribosomal RNA (prRNA) tail and mature ribosomal RNA (mrRNA); (b) contacting a specimen obtained from the sample with the probe or pair of probes in the presence of the antibiotic agent; (c) detecting the relative amounts of probe hybridization to the target sequence in the specimens of (a) and (b); (d) identifying the sample as susceptible to antibiotic treatment if the amount of probe hybridization to the target sequence in step (b) is reduced by at least 80% relative to the amount of probe hybridization to the target sequence in step (a). 14. A method for determining the antibiotic efficacy of a candidate antibiotic agent, the method comprising the steps of: (a) contacting a specimen obtained from the sample with an oligonucleotide probe or pair of probes in the absence of the agent, wherein the probe or pair of probes specifically hybridizes to a target sequence over the full length of the target sequence, wherein the target sequence comprises 25-35 contiguous nucleotides of bacterial ribosomal RNA (rRNA) spanning a splice site between pre-ribosomal RNA (prRNA) tail and mature ribosomal RNA (mrRNA); (b) contacting a specimen obtained from the sample with the probe or pair of probes in the presence of the agent; (c) detecting the relative amounts of probe hybridization to the target sequence in the specimens of (a) and (b); (d) identifying the agent as effective if the amount of probe hybridization to the target sequence in step (b) is reduced by at least 80% relative to the amount of probe hybridization to the target sequence in step (a). 15. A device for detecting pre-rRNA in a bacterial sample, the device comprising an oligonucleotide probe immobilized on a solid support, wherein the oligonucleotide probe is between about 10 to 50 nucleotides in length and is capable of selectively hybridizing to a target sequence over the full length of the target sequence, wherein the target sequence comprises 25-35 contiguous nucleotides of bacterial ribosomal RNA (rRNA) spanning a splice site between pre-ribosomal RNA (prRNA) tail and mature ribosomal RNA (mrRNA). 17. A kit comprising a pair of oligonucleotide probes selected from: (a) E. coli (all enterobacteriaceae) probes:(SEQ ID NO: 5)5'-AAGCCTCACGGTTCATTand(SEQ ID NO: 6)GGCGTTGTAAGGTT;(b) Pseudomonas aeruginosa probes:(SEQ ID NO: 7)5'-AAGCCTCACGGGCAATTand(SEQ ID NO: 8)GGTGTTATATGGTC;(c) Streptococcus pyogenes probes:(SEQ ID NO: 9)AAGTCCTCGAGCTATTand(SEQ ID NO: 10)ATTTCTTTTTGGAT;and(d) Staphylococcus aureus probes(SEQ ID NO: 11)AAGTCTTCGATCGATTand(SEQ ID NO: 12)CATTTATTTTGATT. 18. A method for monitoring the growth rate of a bacterial culture, the method comprising: (a) contacting a specimen obtained from the culture with a probe or pair of probes that specifically hybridizes to a target sequence over the full length of the target sequence, wherein the target sequence comprises 25-35 contiguous nucleotides of bacterial ribosomal RNA (rRNA) spanning a splice site between pre-ribosomal RNA (prRNA) tail and mature ribosomal RNA (mrRNA); (b) detecting the amount of probe hybridization to the target sequence in the specimen of (a) relative to an earlier time point or other control. (c) identifying the culture as growing if the amount of probe hybridization to the target sequence in step (b) is increasing relative to the amount of probe hybridization to the target sequence at the earlier time point.
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法律状态
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(US20150104789) LEGAL DETAILS FOR US2015104789 Actual or expected expiration date=2033-05-03 Legal state=ALIVE Status=PENDING Event publication date=2013-05-03 Event code=US/APP Event indicator=Pos Event type=Examination events Application details Application country=US US14398725 Application date=2013-05-03 Standardized application number=2013US-14398725 Event publication date=2013-05-03 Event code=US/EXMR Event type=Administrative notifications USPTO Examiner Name Primary Examiner: JOHANNSEN, DIANA B Event publication date=2013-05-03 Event code=US/ART Event type=Administrative notifications USPTO Art Group ART=1634 Event publication date=2013-05-03 Event code=US/DK Event type=Examination events Attorney Docket Number Docket Nbr: UCLA.199USWO Customer Nbr: 59612 Event publication date=2013-05-03 Event code=US/ENT Event type=Administrative notifications Business Entity Status: UNDISCOUNTED Event publication date=2013-05-03 Event code=US/AIA Event type=Administrative notifications First Inventor File Indicated: AIA=Yes Event publication date=2014-11-03 Event code=US/PTARDY Event indicator=Pos Event type=Examination events Patent Term Adjustment - Ready for Examination Event publication date=2014-11-03 Event code=US/IDS Event type=Examination events Event type=OAI Information Disclosure Statement Filed Event publication date=2014-11-03 Event code=US/ENTYSTA Event type=Administrative notifications Entity Status Set to Undiscounted Event publication date=2014-11-05 Event code=US/AS Event type=Change of name or address Event type=Reassignment Assignment Owner: THE REGENTS OF THE UNIVERSITY OF CALIFORNIA, CALIF Effective date of the event=2013-07-15 ASSIGNMENT OF ASSIGNORS INTEREST ASSIGNORS:HAAKE, DAVID A. HALFORD, COLIN BABBITT, JANE T. REEL/FRAME:034112/0285 Event publication date=2014-11-05 Event code=US/AS Event type=Change of name or address Event type=Reassignment Assignment Owner: THE REGENTS OF THE UNIVERSITY OF CALIFORNIA, CALIF Effective date of the event=2013-09-23 ASSIGNMENT OF ASSIGNORS INTEREST ASSIGNOR:CHURCHILL, BERNARD M. REEL/FRAME:034112/0271 Event publication date=2014-11-05 Event code=US/AS Event type=Change of name or address Event type=Reassignment Assignment Owner: THE UNITED STATES GOVERNMENT REPRESENTED BY THE DE Effective date of the event=2013-07-15 ASSIGNMENT OF ASSIGNORS INTEREST ASSIGNORS:HAAKE, DAVID A. HALFORD, COLIN BABBITT, JANE T. REEL/FRAME:034112/0285 Event publication date=2015-01-08 Event code=US/APPFILEREC Event type=Administrative notifications Event type=OAO Filing Receipt Event publication date=2015-01-08 Event code=US/M903 Event indicator=Pos Event type=Examination events Event type=OAO Notice of DO/EO Acceptance Mailed Event publication date=2015-01-21 Event code=US/DOCK Event indicator=Pos Event type=Examination events Case Docketed to Examiner in GAU Event publication date=2015-04-16 Event code=US/A1 Event type=Examination events Application published Publication country=US Publication number=US2015104789 Publication stage Code=A1 Publication date=2015-04-16 Standardized publication number=US20150104789 Event publication date=2015-04-16 Event code=US/PGPUBN Event indicator=Pos Event type=Examination events Event type=OAO PG-Pub Issue Notification Event publication date=2015-05-05 Event code=US/AS Event type=Change of name or address Event type=Reassignment Assignment Owner: NATIONAL INSTITUTES OF HEALTH (NIH), U.S. DEPT. OF Effective date of the event=2015-04-21 CONFIRMATORY LICENSE ASSIGNOR:UNIVERSITY OF CALIFORNIA LOS ANGELES REEL/FRAME:035583/0564 Event publication date=2016-02-23 Event code=US/IDS Event type=Examination events Event type=OAI Information Disclosure Statement Filed Event publication date=2017-05-15 Event code=US/CTNF Event type=Examination events Event type=OA Event type=OAO Non-Final Rejection Event publication date=2017-08-23 Event code=US/RECEIPTCOR Event type=Administrative notifications Event type=OAO Filing Receipt - Corrected
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专利类型码
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A1
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国别省市代码
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