DK3108014T3 - Fremgangsmåder og systemer til hurtig påvisning af mikroorganismer under anvendelse af rekombinant bakteriofag - Google Patents
Fremgangsmåder og systemer til hurtig påvisning af mikroorganismer under anvendelse af rekombinant bakteriofag Download PDFInfo
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- DK3108014T3 DK3108014T3 DK15719021.6T DK15719021T DK3108014T3 DK 3108014 T3 DK3108014 T3 DK 3108014T3 DK 15719021 T DK15719021 T DK 15719021T DK 3108014 T3 DK3108014 T3 DK 3108014T3
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- C12N15/09—Recombinant DNA-technology
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- C12Q1/70—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving virus or bacteriophage
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- C12N2795/00—Bacteriophages
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- C12N2795/10011—Details dsDNA Bacteriophages
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- C12N2795/00—Bacteriophages
- C12N2795/00011—Details
- C12N2795/10011—Details dsDNA Bacteriophages
- C12N2795/10111—Myoviridae
- C12N2795/10131—Uses of virus other than therapeutic or vaccine, e.g. disinfectant
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- C12N2795/00—Bacteriophages
- C12N2795/00011—Details
- C12N2795/10011—Details dsDNA Bacteriophages
- C12N2795/10111—Myoviridae
- C12N2795/10141—Use of virus, viral particle or viral elements as a vector
- C12N2795/10143—Use of virus, viral particle or viral elements as a vector viral genome or elements thereof as genetic vector
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- C12N2795/00—Bacteriophages
- C12N2795/00011—Details
- C12N2795/10011—Details dsDNA Bacteriophages
- C12N2795/10211—Podoviridae
- C12N2795/10221—Viruses as such, e.g. new isolates, mutants or their genomic sequences
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- C12N2795/00—Bacteriophages
- C12N2795/00011—Details
- C12N2795/10011—Details dsDNA Bacteriophages
- C12N2795/10211—Podoviridae
- C12N2795/10231—Uses of virus other than therapeutic or vaccine, e.g. disinfectant
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- C12N2795/00—Bacteriophages
- C12N2795/00011—Details
- C12N2795/10011—Details dsDNA Bacteriophages
- C12N2795/10211—Podoviridae
- C12N2795/10241—Use of virus, viral particle or viral elements as a vector
- C12N2795/10243—Use of virus, viral particle or viral elements as a vector viral genome or elements thereof as genetic vector
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Claims (15)
1. Rekombinant bakteriofag, som omfatter et indikatorgen indsat i en late-gen-region af bakteriofagen, hvor indikatorgenet koder for et luciferase-enzym, og hvor indikatorgenet ikke koder for et fusionsprotein, og hvor indikatorproteinproduktet ikke omfatter polypeptider fra den naturlige bakteriofag.
2. Rekombinant bakteriofag ifølge krav 1, hvor indikatorgenet er beliggende i umiddelbar nærhed af et major capsid-gen, og eventuelt hvor transskription af indikatorgenet kontrolleres af en bakteriofag-late-promotor.
3. Rekombinant bakteriofag ifølge krav 1, hvor bakteriofagen er afledt af T7, T4, T4-lignende fag, JG04, JG04-lignende fag eller en anden naturlig bakteriofag med et genom, som udviser mindst 90% homologi med T7, JG04, JG04-lignende fag, T4 eller en anden T4-lignende fag.
4. Bakteriofag ifølge krav 1, hvor ekspression af indikatorgenet under bakteriofagreplika-tion efter infektion af en værtsbakterie resulterer i et opløseligt indikatorproteinprodukt, og eventuelt hvor luciferasen er en af Oplophorus-luciferase, Firefly-luciferase, Lucia-luciferase eller en modificeret luciferase.
5. Fremgangsmåde til påvisning af en mikroorganisme af interesse i en prøve, hvilken fremgangsmåde omfatter følgende trin: inkubering af prøven med en rekombinant bakteriofag, som inficerer mikroorganismen af interesse, hvor den rekombinante bakteriofag omfatter et indikatorgen indsat i en late-gen-region af bakteriofagen, således at ekspression af indikatorgenet under bakteriofagreplikation efter infektion af værtsbakterier resulterer i et opløseligt indikatorproteinprodukt, hvor indikatorgenet koder for et luciferase-enzym, og hvor indikatorgenet ikke koder for et fusionsprotein, og hvor indikatorproteinproduktet ikke omfatter polypeptider fra den naturlige bakteriofag; og påvisning af indikatorproteinproduktet, hvor positiv påvisning af indikatorproteinproduktet indikerer, at mikroorganismen af interesse er til stede i prøven.
6. Fremgangsmåde ifølge krav 5, hvor den påviste mængde indikatordel svarer til mængden af mikroorganismen af interesse, som er til stede i prøven.
7. Fremgangsmåde ifølge krav 5, hvor indikatorgenet er beliggende i umiddelbar nærhed af et major capsid-gen.
8. Fremgangsmåde ifølge krav 5, hvor bakteriofagen er afledt af T7, T4, T4-lignende fag, JG04 eller en anden naturlig bakteriofag med et genom, som udviser mindst 90% homologi med T7, T4 eller en anden T4-lignende fag.
9. Fremgangsmåde ifølge krav 5, hvor bakteriofagkoncentrationen til inkuberingstrinet til inficering af bakterier er større end 1 x 107 plaquedannende enheder pr. milliliter volumen.
10. Fremgangsmåde ifølge krav 5, hvor den rekombinante bakteriofag oprenses ved anvendelse af isopyknisk caesiumchlorid-densitetsgradientcentrifugering inden inkubering med prøven.
11. Fremgangsmåde ifølge krav 5, som endvidere omfatter et trin til indfangning af mikroorganismen fra prøven på en fast bærer inden inkubering med den rekombinante bakteriofag for at inficere bakterier.
12. Fremgangsmåde ifølge krav 11, hvor den faste bærer omfatter en flerbrøndsplade eller et filter.
13. Fremgangsmåde ifølge krav 11, som endvidere omfatter et trin til vask af den indfangede og inficerede mikroorganisme efter tilsætning af bakteriofagen, men inden cellelysis, for at fjerne overskydende bakteriofag og/eller kontaminerende reporterprotein, såsom luciferase.
14. Fremgangsmåde ifølge krav 5, hvor fremgangsmåden kan påvise < 10 celler af mikroorganismen i prøven, og eventuelt hvor påvisning af mikroorganismen af interesse gennemføres på mindre tid end et tidsrum, der er påkrævet til forøgelse af antallet af mikroorganismer 4 gange eller 10 gange ved anvendelse af dyrkning til berigelse, eventuelt hvor den samlede tid, der kræves til påvisning, er mindre end 2 timer.
15. System til hurtig påvisning af en mikroorganisme af interesse i en prøve, hvilket system omfatter: en rekombinant bakteriofag, som inficerer mikroorganismen af interesse, hvor den rekombinante bakteriofag omfatter et indikatorgen indsat i en late-gen-region af bakteriofagen som indikatordelen, således at ekspressionen af indikatorgenet under bakteriofagreplikation efter infektion af værtsbakterier resulterer i et opløseligt indikatorproteinprodukt, hvor indikatorgenet koder for et luciferase-enzym, og hvor indikatorgenet ikke koder for et fusionsprotein, og hvor indikatorprote-inproduktet ikke omfatter polypeptider fra den naturlige bakteriofag; en komponent til inkubering af prøven med den rekombinante bakteriofag; og en komponent til påvisning af indikatordelen.
Applications Claiming Priority (2)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| US201461940959P | 2014-02-18 | 2014-02-18 | |
| PCT/US2015/016415 WO2015126966A1 (en) | 2014-02-18 | 2015-02-18 | Methods and systems for rapid detection of microorganisms using recombinant bacteriophage |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| DK3108014T3 true DK3108014T3 (da) | 2019-02-04 |
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Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| DK15719021.6T DK3108014T3 (da) | 2014-02-18 | 2015-02-18 | Fremgangsmåder og systemer til hurtig påvisning af mikroorganismer under anvendelse af rekombinant bakteriofag |
Country Status (10)
| Country | Link |
|---|---|
| EP (1) | EP3108014B1 (da) |
| JP (2) | JP6767268B2 (da) |
| CN (1) | CN106164259A (da) |
| AU (3) | AU2015219110B2 (da) |
| BR (1) | BR112016018981A2 (da) |
| CA (1) | CA2939564C (da) |
| DK (1) | DK3108014T3 (da) |
| ES (1) | ES2705716T3 (da) |
| MX (2) | MX2016010789A (da) |
| WO (1) | WO2015126966A1 (da) |
Families Citing this family (16)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US10519483B2 (en) | 2012-02-21 | 2019-12-31 | Laboratory Corporation Of America Holdings | Methods and systems for rapid detection of microorganisms using infectious agents |
| US10913934B2 (en) | 2013-02-21 | 2021-02-09 | Laboratory Corporation Of America Holdings | Methods and systems for rapid detection of microorganisms using infectious agents |
| CN115960844A (zh) * | 2016-01-18 | 2023-04-14 | 美国控股实验室公司 | 使用感染原快速检测微生物的方法和系统 |
| CN106995804A (zh) * | 2017-03-20 | 2017-08-01 | 海南大学 | 一种乙酰胆碱酯酶标记的工程化噬菌体快速检测微生物 |
| CN106995805A (zh) * | 2017-03-26 | 2017-08-01 | 海南大学 | 一种溶菌酶标记的工程化噬菌体快速检测微生物 |
| CN106978471A (zh) * | 2017-03-27 | 2017-07-25 | 海南大学 | 一种酰基转移酶标记的工程化噬菌体快速检测微生物 |
| CN107130013A (zh) * | 2017-05-15 | 2017-09-05 | 海南大学 | 一种氨基肽酶标记的工程化噬菌体快速检测微生物 |
| CN111757929A (zh) * | 2018-01-12 | 2020-10-09 | 美国控股实验室公司 | 使用感染因子快速检测沙门氏菌属的方法和系统 |
| CN109874316B (zh) * | 2018-05-25 | 2022-10-11 | 江苏汇先医药技术有限公司 | 用于从样本中富集筛选目标物例如细胞、细菌或生物分子的装置 |
| CN109321457A (zh) * | 2018-10-17 | 2019-02-12 | 深圳市深研生物科技有限公司 | 一种制备细胞的设备及其应用 |
| EP3918331A1 (en) * | 2019-01-29 | 2021-12-08 | Laboratory Corporation of America Holdings | Methods and systems for the rapid detection of listeria using infectious agents |
| WO2020257502A1 (en) | 2019-06-21 | 2020-12-24 | Laboratory Corporation Of America Holdings | Methods for producing mutant bacteriophages for the detection of listeria |
| CA3147173A1 (en) | 2019-08-26 | 2021-03-04 | Stephen Erickson | Devices and methods for detecting microorganisms using recombinant reproduction-deficient indicator bacteriophage |
| JP2022547218A (ja) | 2019-09-11 | 2022-11-10 | ラボラトリー コーポレイション オブ アメリカ ホールディングス | 指標サブユニットを発現するように組換え感染病原体を使用して微生物を迅速に検出するための方法およびシステム |
| EP4165412A1 (en) * | 2020-06-15 | 2023-04-19 | Laboratory Corporation of America Holdings | Methods for the diagnosis and treatment of biofilm-related infections |
| US20220214343A1 (en) * | 2020-11-25 | 2022-07-07 | Laboratory Corporation Of America Holdings | Methods and Systems for the Detection of Microorganisms Using Infectious Agents |
Family Cites Families (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPH10179161A (ja) * | 1996-12-26 | 1998-07-07 | Tomono Agurika:Kk | 細菌検出法 |
| WO2006075996A2 (en) * | 2004-04-07 | 2006-07-20 | The Government Of The United States Of America, As Represented By The Secretary, Department Of Health And Human Services | Reporter plasmid phage packaging system for detection of bacteria |
| CN101680041A (zh) * | 2007-04-18 | 2010-03-24 | 康奈尔大学 | 微生物检测方法和仪器 |
| JP2015510598A (ja) * | 2012-02-21 | 2015-04-09 | ラボラトリー コーポレイション オブ アメリカ ホールディングス | 微生物の検出のための方法およびシステム |
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2015
- 2015-02-18 AU AU2015219110A patent/AU2015219110B2/en active Active
- 2015-02-18 MX MX2016010789A patent/MX2016010789A/es active IP Right Grant
- 2015-02-18 BR BR112016018981A patent/BR112016018981A2/pt not_active Application Discontinuation
- 2015-02-18 WO PCT/US2015/016415 patent/WO2015126966A1/en active Application Filing
- 2015-02-18 CA CA2939564A patent/CA2939564C/en active Active
- 2015-02-18 EP EP15719021.6A patent/EP3108014B1/en active Active
- 2015-02-18 JP JP2016569585A patent/JP6767268B2/ja active Active
- 2015-02-18 CN CN201580009143.1A patent/CN106164259A/zh active Pending
- 2015-02-18 DK DK15719021.6T patent/DK3108014T3/da active
- 2015-02-18 ES ES15719021T patent/ES2705716T3/es active Active
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2016
- 2016-08-18 MX MX2020010692A patent/MX2020010692A/es unknown
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- 2020-09-17 JP JP2020156264A patent/JP7210518B2/ja active Active
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2021
- 2021-09-20 AU AU2021236442A patent/AU2021236442B9/en active Active
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- 2024-08-07 AU AU2024205589A patent/AU2024205589A1/en active Pending
Also Published As
| Publication number | Publication date |
|---|---|
| AU2021236442A1 (en) | 2021-10-14 |
| CA2939564C (en) | 2024-01-16 |
| WO2015126966A1 (en) | 2015-08-27 |
| ES2705716T3 (es) | 2019-03-26 |
| JP6767268B2 (ja) | 2020-10-21 |
| AU2021236442B9 (en) | 2024-05-30 |
| AU2024205589A1 (en) | 2024-09-12 |
| MX2020010692A (es) | 2020-11-06 |
| AU2015219110B2 (en) | 2021-07-01 |
| EP3108014B1 (en) | 2018-12-05 |
| JP2020202872A (ja) | 2020-12-24 |
| AU2021236442B2 (en) | 2024-05-09 |
| CN106164259A (zh) | 2016-11-23 |
| EP3108014A1 (en) | 2016-12-28 |
| AU2015219110A1 (en) | 2016-09-08 |
| CA2939564A1 (en) | 2015-08-27 |
| BR112016018981A2 (pt) | 2017-10-10 |
| JP2017511702A (ja) | 2017-04-27 |
| JP7210518B2 (ja) | 2023-01-23 |
| MX2016010789A (es) | 2016-11-08 |
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