WO2012096646A9 - Détection directe de l'arn non amplifié du virus de l'hépatite c à l'aide de nanoparticules d'or non modifiées - Google Patents

Détection directe de l'arn non amplifié du virus de l'hépatite c à l'aide de nanoparticules d'or non modifiées Download PDF

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Publication number
WO2012096646A9
WO2012096646A9 PCT/US2011/020676 US2011020676W WO2012096646A9 WO 2012096646 A9 WO2012096646 A9 WO 2012096646A9 US 2011020676 W US2011020676 W US 2011020676W WO 2012096646 A9 WO2012096646 A9 WO 2012096646A9
Authority
WO
WIPO (PCT)
Prior art keywords
hcv
hepatitis
rna
sample
oligotargeter
Prior art date
Application number
PCT/US2011/020676
Other languages
English (en)
Other versions
WO2012096646A1 (fr
Inventor
Hassan Mohamed EL-SAID AZZAZY EL-BADAWY
Sherif Mohamed SHAWKY ABDUO
Original Assignee
The American University In Cairo
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by The American University In Cairo filed Critical The American University In Cairo
Priority to PCT/US2011/020676 priority Critical patent/WO2012096646A1/fr
Priority to EP11855330.4A priority patent/EP2663649A4/fr
Priority to MYPI2013002624A priority patent/MY166402A/en
Publication of WO2012096646A1 publication Critical patent/WO2012096646A1/fr
Publication of WO2012096646A9 publication Critical patent/WO2012096646A9/fr

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Classifications

    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
    • C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
    • C12Q1/70—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving virus or bacteriophage
    • C12Q1/701—Specific hybridization probes
    • C12Q1/706—Specific hybridization probes for hepatitis
    • C12Q1/707—Specific hybridization probes for hepatitis non-A, non-B Hepatitis, excluding hepatitis D

Definitions

  • dsDNA double-stranded DNA
  • dsDNA double-stranded DNA
  • nanoparticles exhibit a red color and aggregated gold nanoparticles a blue color, where a blue color is indicative of the presence of HCV RNA in a sample that is complementary to the oligotargeter is conjugated to an FAM dye or other fluorescent dye or fluorophore whose emission can be quenched by gold nanoparticles; and the presence of HCV is detected by the emission of fluorescence.
  • RIBA Recombinant immune blot assays.
  • AuNPs Gold Nanoparticles.
  • a tissue sample may be used in the assay or processed for use in the assay, for example, by a conventional method used to extract HCV or HCV nucleic acids from the sample.
  • purified HCV describes HCV which has been isolated from the host tissues or fluids in which the virus is normally associated, isolated from a tissue cell culture, or separated from other types of microorganisms, such as bacteria or other viruses. Techniques for isolating HCV are known to those of skill in the art.
  • modified oligotargeter describes an oligotargeter sequence that
  • modifications to increase nuclease resistance of the oligotargeter include the following: (a) phosphothioate modified sequence (where one of the oxygen on the phosphate of phosphodiester bond is replaced with a sulphur atom); (b) 3'-propryl group (C3 spacer, adding a propyl group at the 3' end); and (c) Inverted end (3'-3' linkage), though other modifications known to those in the art may also be employed.
  • HCV RNA refers to RNA from a HCV viral genome or synthetic RNA corresponding to genomic sequences, fragments thereof, transcripts thereof, or modified or mutant sequences derived HCV sequences. It also encompass modified or mutated HCV genomic sequences, such as variants containing one or more single nucleotide polymorphisms, or more generally, those having a sequence containing 1 , 2, 3, 4, 5 or more insertions, deletions, transpositions, or substitutions to a genomic HCV sequence.
  • each gold nanoparticle may be stabilized from salt-induced aggregation when it is covered by about 12 oligotargeter molecules.
  • One example of a suitable ratio of target, oligotargeter and gold nanoparticles would be 7 microliters of extracted RNA of unknown or variable concentration in combination with ⁇ ⁇ oligotargeter which is admixed with 10 nM gold nanoparticles.
  • RNA detection during the acute phase infection as optimal time after infection to initiate therapy (8- 12 weeks), optimal treatment duration (24 weeks) and ribavirin is not required for optimal responses during acute infection, thus reducing the risk of major adverse effects.
  • optimal time after infection to initiate therapy 8- 12 weeks
  • optimal treatment duration 24 weeks
  • ribavirin is not required for optimal responses during acute infection, thus reducing the risk of major adverse effects.

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  • Health & Medical Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Organic Chemistry (AREA)
  • Wood Science & Technology (AREA)
  • Engineering & Computer Science (AREA)
  • Zoology (AREA)
  • Communicable Diseases (AREA)
  • Immunology (AREA)
  • Proteomics, Peptides & Aminoacids (AREA)
  • Molecular Biology (AREA)
  • Biotechnology (AREA)
  • Microbiology (AREA)
  • Physics & Mathematics (AREA)
  • Biophysics (AREA)
  • Analytical Chemistry (AREA)
  • Virology (AREA)
  • Biochemistry (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • General Engineering & Computer Science (AREA)
  • General Health & Medical Sciences (AREA)
  • Genetics & Genomics (AREA)
  • Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)

Abstract

L'invention concerne une trousse pour un essai colorimétrique à base de nanoparticules d'or pour un ARN du virus de l'hépatite C qui détecte l'ARN du VHC non amplifié dans des échantillons cliniques à l'aide de AuNP non modifiées et de polynucléotides ciblant les oligonucléotides qui se lient à l'ARN du VHC. L'invention concerne un procédé de détection du virus de l'hépatite C comprenant la mise en contact d'un échantillon suspecté de contenir le virus de l'hépatite C avec un polynucléotide qui se lie à l'ARN du virus de l'hépatite C et des nanoparticules d'or, la détection de l'agrégation de nanoparticules et la détection du virus de l'hépatite C dans l'échantillon lorsque que les nanoparticules s'agrègent (la couleur de la solution devient bleue) en comparaison à un échantillon témoin ou négatif ne contenant pas le virus lorsque les nanoparticules ne s'agrègent pas (la couleur de la solution reste rouge).
PCT/US2011/020676 2011-01-10 2011-01-10 Détection directe de l'arn non amplifié du virus de l'hépatite c à l'aide de nanoparticules d'or non modifiées WO2012096646A1 (fr)

Priority Applications (3)

Application Number Priority Date Filing Date Title
PCT/US2011/020676 WO2012096646A1 (fr) 2011-01-10 2011-01-10 Détection directe de l'arn non amplifié du virus de l'hépatite c à l'aide de nanoparticules d'or non modifiées
EP11855330.4A EP2663649A4 (fr) 2011-01-10 2011-01-10 Détection directe de l'arn non amplifié du virus de l'hépatite c à l'aide de nanoparticules d'or non modifiées
MYPI2013002624A MY166402A (en) 2011-01-10 2011-01-10 Direct detection of unamplified hepatitis c virus rna using unmodified gold nanoparticles

Applications Claiming Priority (1)

Application Number Priority Date Filing Date Title
PCT/US2011/020676 WO2012096646A1 (fr) 2011-01-10 2011-01-10 Détection directe de l'arn non amplifié du virus de l'hépatite c à l'aide de nanoparticules d'or non modifiées

Publications (2)

Publication Number Publication Date
WO2012096646A1 WO2012096646A1 (fr) 2012-07-19
WO2012096646A9 true WO2012096646A9 (fr) 2012-08-30

Family

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Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/US2011/020676 WO2012096646A1 (fr) 2011-01-10 2011-01-10 Détection directe de l'arn non amplifié du virus de l'hépatite c à l'aide de nanoparticules d'or non modifiées

Country Status (3)

Country Link
EP (1) EP2663649A4 (fr)
MY (1) MY166402A (fr)
WO (1) WO2012096646A1 (fr)

Families Citing this family (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JP6217495B2 (ja) 2014-03-31 2017-10-25 ブラザー工業株式会社 圧電アクチュエータ
CN113151584A (zh) * 2021-01-11 2021-07-23 南通大学 一种SARS-CoV-2检测试剂盒及检测方法
CN115961060A (zh) * 2022-08-22 2023-04-14 中国海洋大学 基于纳米金的等温扩增检测结核分枝杆菌的引物探针组、试剂盒、方法及应用

Family Cites Families (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
NL1002781C1 (nl) * 1996-04-03 1997-10-06 Amsterdam Support Diagnostics Isolatie en/of amplificatie van hepatitis-C-virus-(HCV) -nucleïnezuren uit monsters waarvan vermoed wordt dat zij HCV bevatten.
US6083482A (en) * 1999-05-11 2000-07-04 Icn Pharmaceuticals, Inc. Conformationally locked nucleosides and oligonucleotides
CA2552949C (fr) * 2004-01-07 2012-10-02 Third Wave Technologies, Inc. Determination d'un genotype du virus de l'hepatite c
WO2009131661A2 (fr) * 2008-04-21 2009-10-29 Dicerna Pharmaceuticals, Inc. Procédés et compositions pour inhiber spécifiquement le virus de l'hépatite c (vhc) au moyen d'arn bicaténaire

Also Published As

Publication number Publication date
WO2012096646A1 (fr) 2012-07-19
EP2663649A4 (fr) 2014-08-27
EP2663649A1 (fr) 2013-11-20
MY166402A (en) 2018-06-25

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