Concentration and enrichment of microbial cells and microbial nucleic acids from bodily fluids
원문보기
IPC분류정보
국가/구분
United States(US) Patent
등록
국제특허분류(IPC7판)
C12Q-001/68
C12Q-001/24
C12P-019/34
출원번호
US-0671741
(2008-08-01)
등록번호
US-8481265
(2013-07-09)
국제출원번호
PCT/CA2008/001414
(2008-08-01)
§371/§102 date
20100727
(20100727)
국제공개번호
WO2009/015484
(2009-02-05)
발명자
/ 주소
Peytavi, Régis
Huletsky, Ann
Belley-Montfort, Lucile
Martineau, Isabelle
출원인 / 주소
Universite Laval
대리인 / 주소
Clark & Elbing LLP
인용정보
피인용 횟수 :
0인용 특허 :
9
초록▼
The present invention relates to a method for isolating microorganisms and/or microorganisms nucleic acids from a bodily fluid that may comprise or may be suspected to comprise microorganisms and/or host cells and/or host cells debris. Microorganisms nucleic acids may further be isolated by lysing t
The present invention relates to a method for isolating microorganisms and/or microorganisms nucleic acids from a bodily fluid that may comprise or may be suspected to comprise microorganisms and/or host cells and/or host cells debris. Microorganisms nucleic acids may further be isolated by lysing the isolated microorganisms. The present invention also relates to a method for detecting microorganisms in a bodily fluid. The present invention further relates to a saponin formulation and its use.
대표청구항▼
1. A method for isolating bacteria and/or bacterial nucleic acids from a blood sample comprising or suspected of comprising bacteria, the method comprising: a. contacting the blood sample with a saponin formulation, wherein the saponin is a filtered autoclaved-treated saponin solution (FATS) or a he
1. A method for isolating bacteria and/or bacterial nucleic acids from a blood sample comprising or suspected of comprising bacteria, the method comprising: a. contacting the blood sample with a saponin formulation, wherein the saponin is a filtered autoclaved-treated saponin solution (FATS) or a heated filtered autoclaved-treated saponin solution (HFATS) present at a final concentration of 40 mg/ml to 100 mg/ml; andb. obtaining isolated bacteria; whereby the isolated bacteria comprises nucleic acids. 2. The method of claim 1, wherein the isolated bacterial are viable and/or metabolically active. 3. The method of claim 1, wherein the final concentration of saponin is from 75 mg/mL to 100 mg/mL. 4. The method of claim 1, wherein said saponin formulation comprises saponin selected from the group consisting of steroidal saponin, triterpenoid saponin and combination thereof. 5. The method of claim 1, wherein the saponin formulation is a triterpenoid saponin formulation. 6. The method of claim 1, wherein the blood sample is a human blood sample. 7. The method of claim 1, wherein obtaining isolated microorganisms comprises at least one centrifugation and or filtration step. 8. The method of claim 1, wherein said method provides for isolation of bacteria present at a concentration as low as 0.2 to 22 CFU per mL of blood. 9. The method of claim 1, wherein the bacteria are selected from the group consisting of Enterococcus faecalis, Enterococcus faeciwn, Escherichia coli, Haemophilus influenzae, Klebsiella oxytoca, Klebsiella pneumoniae, Proteus mirabilis, Pseudomonas aeruginosa, Serratia marcescens, Staphylococcus aureus, Staphylococcus epidermidis, Staphylococcus haemolyticus, Staphylococcus hominis, Staphylococcus warneri, Stenotrophomonas maltophilia, Streptococcus agalactiae, Streptococcus anginosus, Streptococcus dysgalactiae, Streptococcus mutans, Streptococcus pneumoniae, Streptococcus pyogenes, Streptococcus sanguinis and combination thereof. 10. The method of claim 1, wherein the bacteria are bacteria which may cause a bloodstream infection. 11. The method of claim 1, whereby the isolated bacteria are substantially free of amplification inhibitors. 12. The method of claim 1, further comprising lysing the isolated bacteria and isolating therefrom nucleic acids. 13. The method of claim 12, further comprising amplifying said nucleic acids. 14. A method for detecting bacteria in a blood sample, comprising: a. contacting a blood sample with a saponin formulation to a final concentration of saponin in a range of 75 mg/mL to 100 mg/mL, wherein said saponin formulation is a heated filtered autoclaved-treated saponin (HFATS) solution;b. isolating bacteria having nucleic acids by using at least one centrifugation and/or at least one filtration step; andc. assessing the presence of isolated bacteria having nucleic acids. 15. The method of claim 14, wherein assessing the presence of isolated bacteria comprises amplifying nucleic acids from said bacteria. 16. The method of claim 14, wherein said bacteria are present in the blood sample at a concentration as low as 0.2 to 22 CFU per mL of blood. 17. A method for obtaining nucleic acids from bacteria present in a blood sample, the method comprising: a. contacting the blood sample with a saponin formulation, wherein saponin is a filtered autoclaved-treated saponin solution (FATS) or a heated filtered autoclaved-treated saponin solution (HFATS) present at a final concentration of 40 mg/ml to 100 mg/ml;b. obtaining isolated bacteria comprising nucleic acids; andc. lysing the isolated bacteria for obtaining therefrom nucleic acids. 18. The method of claim 17, wherein obtaining isolated bacteria comprises at least one centrifugation and/or at least one filtration step. 19. The method of claim 18, further comprising amplifying nucleic acids from said bacteria for identification and/or diagnostic purposes. 20. A method for diagnosing a blood infection in a subject in need thereof comprising: a) contacting a blood sample from said subject with a saponin formulation using techniques and conditions where microorganisms comprising nucleic acids are isolated from host cells and host cell debris, wherein saponin is a filtered autoclaved-treated saponin solution (FATS) or a heated filtered autoclaved-treated saponin solution (HFATS) present at a final concentration of 40 mg/ml to 100 mg/ml; andb) assessing presence or absence of bacteria comprising nucleic acids; whereby detection of bacteria comprising nucleic acids is indicative of an infection associated with the bacteria detected.
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이 특허에 인용된 특허 (9)
McAllister Diane L. (San Diego CA) Clark Kathleen A. (Cardiff-By-The-Sea CA), Amplification primers and nucleic acid probes targeted to coccidioides immitis nucleic acid.
Fletcher James C. Administrator of the National Aeronautics and Space Administration ; with respect to an invention of ( Baltimore MD) Chappelle Emmett W. (Baltimore MD) Picciolo Grace L. (Tantallon , Application of luciferase assay for ATP to antimicrobial drug susceptibility.
Dorn Gordon L. (6647 Lakewood Blvd. Dallas TX 75214), Method for improving quantitative recovery of microorganisms from specimens containing blood components.
Berger Dolores M. (Baltimore MD) Goldenbaum Paul E. (Towson MD) Tice Gregory (Lutherville MD), Method for reducing blood carbon dioxide background in bacterial media by the addition of micelles of saponin and a phos.
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