WO2013064834A1 - Essai de la dengue - Google Patents

Essai de la dengue Download PDF

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Publication number
WO2013064834A1
WO2013064834A1 PCT/GB2012/052731 GB2012052731W WO2013064834A1 WO 2013064834 A1 WO2013064834 A1 WO 2013064834A1 GB 2012052731 W GB2012052731 W GB 2012052731W WO 2013064834 A1 WO2013064834 A1 WO 2013064834A1
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WO
WIPO (PCT)
Prior art keywords
seq
ttt
deng
probe
ctc
Prior art date
Application number
PCT/GB2012/052731
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English (en)
Inventor
Ben Cobb
Original Assignee
Epistem Limited
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 Epistem Limited filed Critical Epistem Limited
Publication of WO2013064834A1 publication Critical patent/WO2013064834A1/fr

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    • 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
    • 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
    • C12Q1/68—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
    • C12Q1/6844—Nucleic acid amplification reactions
    • C12Q1/686—Polymerase chain reaction [PCR]

Definitions

  • the present invention relates to an assay for detection of dengue virus in samples.
  • Dengue viruses belong to the family Flaviviridae, genus Flavivirus. The viruses occur as four antigenically distinct serotypes; 1 , 2, 3 & 4. Infection with one type usually gives lifelong immunity to that type, but only short-term immunity to the others. Subsequent infection with a different type increases the risk of severe complications. Dengue is transmitted by several species of mosquito, and the disease continues to be a major public health problem in tropical and subtropical areas. Initial infection with dengue generally leads to a mild, self-limiting febrile illness (dengue fever), but the more severe form of the disease is responsible for a high mortality rate, primarily among children and involves vascular and hemostatic abnormalities (dengue hemorrhagic fever- dengue shock syndrome [DHFDSS]).
  • DHFDSS vascular and hemostatic abnormalities
  • Definitive diagnosis of dengue serotype is complex. Testing can be done typically by nucleic acid analysis by PCR, or by detection of specific antibodies to the viral type.
  • Dengue virus contains a positive-sense single-stranded RNA genome of approximately 1 1 kb. There is very little by way of conserved sequence within the genome, which limits the scope for a universal PCR genotyping method. Most tests are based on Lanciotti et al (Rapid Detection and Typing of Dengue Viruses from Clinical Samples by Using Reverse Transcriptase-Polymerase Chain Reaction, Journal of Clinical Microbiology, Mar. 1992, p. 545-551 ).
  • a method for determining the serotype of a dengue virus within a sample comprising:
  • the degree of similarity between the probe and the amplified portion is indicative of the viral serotype.
  • the method thus relies on the absence of conserved sequences within the genome, as the amplified sequence will be different depending on the viral serotype.
  • a single primer pair (or a small primer set) is able to amplify the sequence.
  • a single probe can then be used to hybridise to any amplified sequence.
  • the melting temperature of the probe will differ depending on the degree of homology between the probe and target. Determining this Tm will allow the serotype to be determined.
  • a single primer pair and single probe may be used to determine the serotype of the dengue virus. Effectively, the present invention allows the use of a single assay whereas prior art assays require multiple assays, one for each serotype.
  • the method may determine the serotype of dengue virus types 1 , 2, 3, and 4. As such, the amplified portion will differ between each of these types.
  • the amplification step may comprise PCR, or RT-PCR.
  • primer combinations are used which will amplify a sequence from any of the viral serotypes.
  • a preferred region for amplification is within the first 200 bp of the Dengue complete genome, and more preferably between bases 3 to 170.
  • the primers preferably amplify a portion of the dengue sequence which is at least 50, 75, 90, 100, 150 nucleotides in length.
  • the primer set comprises at least one, and preferably all, forward primer(s) selected from SEQ ID NO: 1 to SEQ ID NO: 3:
  • the primer set preferably also comprises the reverse primer SEQ ID NO: 4:
  • An alternative downstream site at base 3 may be used to amplify using two primers.
  • the primer set preferably comprises at least one forward primer selected from SEQ ID NO: 5 and 6.
  • the primer set may comprise at least one reverse primer selected from SEQ ID NO: 7, 8, and 9.
  • the nucleic acid probe is preferably at least 10, 1 1 , 12, 13, 14, 15, 16, 17, 18 nucleotides in length.
  • a particularly preferred nucleic acid probe comprises, or consists of, the sequence AAC CAA CGA AAA AAG GCG (SEQ ID NO: 10).
  • Alternative probes which may be used include TTT GAA TAG AGA GCA GAT CT (SEQ ID NO: 1 1 ); or TAG AGA GCA GAT CTC TGA TGA A (SEQ ID NO: 12). Each of these latter two binds within the region to which the forward primer DENG-F 73 (SEQ ID NO: 1 ) hybridises.
  • the method may optionally comprise contacting the amplified portion with a second nucleic acid probe designed to hybridise to at least a fragment of the amplified portion. Combining two probes can give added accuracy in the call rate of the different subtypes.
  • the probe may be labelled.
  • the probe may include a fluorescent or a radioactive label, or may be labelled with a ligand to which a secondary probe may bind.
  • the probe is labelled with a fluorescent label, and preferably also the label generates a differential signal depending on whether the probe has hybridised to a target strand (that is, the probe is part of a double stranded nucleic acid) or not (the probe is single stranded).
  • a preferred probe is a HyBeacon® probe (see, for example, Mol Cell Probes.
  • the determining step may comprise varying the temperature of the sample, and determining the temperature at which the probe hybridises to the amplified portion. Conveniently this is achieved using labelled probes which fluoresce when in double stranded form; this allows rapid detection of the melting temperature simply by measuring fluorescence levels.
  • the temperature may be raised to melt any double stranded nucleic acids, and gradually lowered to determine the temperature at which hybridisation takes place. Alternatively, the temperature may first be lowered to allow hybridisation, and gradually increased to allow melting of hybridised probe.
  • the method may further comprise the steps of amplifying and detecting control nucleic acids.
  • the sample may be for example blood, sputum, serum, urine, mucus.
  • a further aspect of the invention provides a kit for detecting and typing dengue virus in a sample, the kit comprising
  • nucleic acid primer set for amplifying a portion of the dengue viral genome, the amplified portion differing in sequence between the viral types to be detected; and b) a nucleic acid probe for hybridising to at least a fragment of the amplified portion.
  • the kit may further comprise instructions for use.
  • the primer set preferably comprises one or more of DENG-F 73, SEQ ID NO: 1 (5'- CAGTTTTTTATTAGAGAGCAGATCTCTG-3') and DREVJ 42, SEQ ID NO: 4 (5'- CGGTTTCTCMCGCGTTTCAGCATATTGA-3'). These primers in combination give a 97 bp amplification product.
  • the primer set in certain embodiments comprises one or more, preferably all, of forward primers selected from SEQ ID NO: 1 , 2, and 3.
  • the primer set may comprise one or more forward primers selected from SEQ ID NO: 5 and 6; and/or one or more reverse primers selected from SEQ ID NO: 7, 8, and 9.
  • the probe is preferably labelled; more preferably labelled in a manner such that the label generates a differential signal depending on whether the probe has hybridised to a target strand.
  • the probe may be selected from nucleic acids having the sequence AAC CAA CGA AAA AAG GCG (SEQ ID NO: 10); TTT GAA TAG AGA GCA GAT CT (SEQ ID NO: 1 1 ); or TAG AGA GCA GAT CTC TGA TGA A (SEQ ID NO: 12).
  • a further aspect of the invention provides a nucleic acid primer set comprising DENG- F_73, SEQ ID NO: 1 (5'-CAGTTTTTTATTAGAGAGCAGATCTCTG-3') and DREVJ 42, SEQ ID NO: 4 (5'-CGGTTTCTCMCGCGTTTCAGCATATTGA-3').
  • the primer set in certain embodiments comprises one or more, preferably all, of forward primers selected from SEQ ID NO: 1 , 2, and 3.
  • the primer set may comprise one or more forward primers selected from SEQ ID NO: 5 and 6; and/or one or more reverse primers selected from SEQ ID NO: 7, 8, and 9.
  • Figure 1 shows an alignment of the nucleic acid sequences of a portion of the dengue virus types 1 , 2, 3, and 4, together with a consensus sequence.
  • the positions of DENG-F 73 (DFWD) and DREV 142 (DREV) primers are marked, together with the location of the target of probe SEQ ID NO: 10.
  • Figure 2 shows the results of a melt curve analysis in which the melting temperature of the probe having SEQ ID NO 10 against samples of dengue virus types 1 , 2, 4.1 and 4.2.
  • sequences shown in Figure 1 are: consensus sequence; type 1 (HQ891313.1 & HQ891314.1 ); type 2 (HQ541786.1 & JF730052.1 ); type 3 (JN000937.1 & JN093514.1 ); type 4 (GU289913.1 & AF289029.1 ).
  • SEQ ID NO 10 The location of a preferred probe, SEQ ID NO 10, is also marked on Figure 1 .
  • a sample is first obtained from a patient suspected of having a dengue viral infection.
  • the sample may be for example a body fluid such as sputum, pus, blood, or serum.
  • the sample may then undergo any necessary sample preparation and processing; although in most cases it may be possible to conduct the assay directly on untreated samples, making it suitable for rapid use in the field.
  • the sample is then subjected to RT-PCT using the forward and reverse primers DFWD and DREV.
  • a reaction mix including dNTPs, reverse transcriptase, and random primers is first used to initiate reverse transcription of viral RNA to give cDNA.
  • the cDNA is then amplified using PCR with the DFWD and DREV primers; a second reaction mix including dNTPs and DNA polymerase is used.
  • the skilled person will be aware of typical reaction mixes for carrying out RT-PCR, and of suitable reaction conditions for the PCR amplification.
  • a 97 bp fragment is amplified from all dengue viral types.
  • a detection reaction mix including a universal probe.
  • the probe is SEQ ID NO 10, AAC CAA CGA AAA AAG GCG.
  • the probe is designed so as to hybridise to a portion of the amplified sequence from all viral types, although the variability in sequence means that the precise melting temperature will differ depending on the type.
  • the detection mix plus amplified fragment is then subjected to a melt curve analysis. The results of one such analysis are given in Figure 2, which illustrates typical curves for viral types 1 , 2, 4.1 , and 4.2. It can be seen from the Figure that the peaks for each type can be clearly distinguished, with the types having different melting temperatures. In this way, the type of dengue virus present in the sample can be readily and rapidly determined, without the need to conduct separate assays for each type.

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  • Chemical & Material Sciences (AREA)
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Abstract

La présente invention concerne un essai pour la détection du virus de la dengue dans des échantillons. Une paire d'amorces est prévue, laquelle amplifie une séquence à partir de chacun des quatre sérotypes du virus de la dengue, ainsi qu'une sonde unique qui s'hybride à une température différente à la séquence amplifiée, en fonction du sérotype. De cette façon, un ensemble unique de sondes et d'amorces peut être utilisé pour effectuer une distinction entre les divers sérotypes.
PCT/GB2012/052731 2011-11-04 2012-11-02 Essai de la dengue WO2013064834A1 (fr)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
GBGB1119028.7A GB201119028D0 (en) 2011-11-04 2011-11-04 Dengue assay
GB1119028.7 2011-11-04

Publications (1)

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WO2013064834A1 true WO2013064834A1 (fr) 2013-05-10

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Cited By (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2014055746A1 (fr) * 2012-10-04 2014-04-10 The Board Of Trustees Of The Leland Stanford Junior University Procédés et réactifs permettant la détection, la quantification et la sérotypie du virus de la dengue
KR20150143425A (ko) * 2013-07-05 2015-12-23 (주)바이오니아 뎅기 바이러스 특이적 siRNA, 그러한 siRNA 를 포함하는 이중나선 올리고 RNA 구조체 및 이를 포함하는 뎅기 바이러스 증식 억제용 조성물
CN105441588A (zh) * 2015-12-21 2016-03-30 深圳市生科源技术有限公司 登革热ⅰ、ⅱ、ⅲ、ⅳ型rt-pcr一步mix检测试剂盒及其检测方法
CN110760614A (zh) * 2019-04-29 2020-02-07 杭州市上城区疾病预防控制中心 一种登革热病毒核酸快速检测、分型以及溯源区分的方法

Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US6855521B2 (en) 1999-12-01 2005-02-15 The United States Of America As Represented By The Secretary Of The Navy Serotype and dengue group specific flurogenic probe based PCR (TaqMan) assays against the respective C and NS5 genomic and 3′ non-coding regions of dengue virus
WO2011026139A1 (fr) * 2009-08-31 2011-03-03 Gen-Probe Incorporated Essai du virus de la dengue

Patent Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US6855521B2 (en) 1999-12-01 2005-02-15 The United States Of America As Represented By The Secretary Of The Navy Serotype and dengue group specific flurogenic probe based PCR (TaqMan) assays against the respective C and NS5 genomic and 3′ non-coding regions of dengue virus
WO2011026139A1 (fr) * 2009-08-31 2011-03-03 Gen-Probe Incorporated Essai du virus de la dengue

Non-Patent Citations (7)

* Cited by examiner, † Cited by third party
Title
C. L.H. LO ET AL: "One-Step Rapid Reverse Transcription-PCR Assay for Detecting and Typing Dengue Viruses with GC Tail and Induced Fluorescence Resonance Energy Transfer Techniques for Melting Temperature and Color Multiplexing", CLINICAL CHEMISTRY, vol. 53, no. 4, 1 April 2007 (2007-04-01), pages 594 - 599, XP055050469, ISSN: 0009-9147, DOI: 10.1373/clinchem.2006.077446 *
CHIEN LI-JUNG ET AL: "Development of real-time reverse transcriptase PCR assays to detect and serotype dengue viruses", JOURNAL OF CLINICAL MICROBIOLOGY, AMERICAN SOCIETY FOR MICROBIOLOGY, WASHINGTON, DC, US, vol. 44, no. 4, 1 April 2006 (2006-04-01), pages 1295 - 1304, XP009139728, ISSN: 0095-1137, DOI: 10.1128/JCM.44.4.1295 1304.2006 *
CHUTINIMITKUL S ET AL: "Dengue typing assay based on real-time PCR using SYBR Green I", JOURNAL OF VIROLOGICAL METHODS, ELSEVIER BV, NL, vol. 129, no. 1, 1 October 2005 (2005-10-01), pages 8 - 15, XP027667404, ISSN: 0166-0934, [retrieved on 20051001] *
CONCEICAO T M ET AL: "A real-time PCR procedure for detection of dengue virus serotypes 1, 2, and 3, and their quantitation in clinical and laboratory samples", JOURNAL OF VIROLOGICAL METHODS, ELSEVIER BV, NL, vol. 163, no. 1, 1 January 2010 (2010-01-01), pages 1 - 9, XP026788538, ISSN: 0166-0934, [retrieved on 20091012], DOI: 10.1016/J.JVIROMET.2009.10.001 *
FRENCH DJ; ARCHARD CL; ANDERSEN MT; MCDOWELL DG: "Ultra-rapid DNA analysis using HyBeacon probes and direct PCR amplification from saliva", MOL CELL PROBES., vol. 16, no. 5, October 2002 (2002-10-01), pages 319 - 26
LANCIOTTI ET AL.: "Rapid Detection and Typing of Dengue Viruses from Clinical Samples by Using Reverse Transcriptase-Polymerase Chain Reaction", JOURNAL OF CLINICAL MICROBIOLOGY, March 1992 (1992-03-01), pages 545 - 551
NAZE F ET AL: "Simultaneous detection and quantitation of Chikungunya, Dengue and West Nile viruses by multiplex RT-PCR assays and Dengue virus typing using High Resolution Melting", JOURNAL OF VIROLOGICAL METHODS, ELSEVIER BV, NL, vol. 162, no. 1-2, 1 December 2009 (2009-12-01), pages 1 - 7, XP026682311, ISSN: 0166-0934, [retrieved on 20090317], DOI: 10.1016/J.JVIROMET.2009.03.006 *

Cited By (9)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2014055746A1 (fr) * 2012-10-04 2014-04-10 The Board Of Trustees Of The Leland Stanford Junior University Procédés et réactifs permettant la détection, la quantification et la sérotypie du virus de la dengue
US9725774B2 (en) 2012-10-04 2017-08-08 The Board Of Trustees Of The Leland Stanford Junior University Methods and reagents for detection, quantitation, and serotyping of dengue viruses
US10093994B2 (en) 2012-10-04 2018-10-09 The Board Of Trustees Of The Leland Stanford Junior University Methods and reagents for detection, quantitation, and serotyping of dengue viruses
KR20150143425A (ko) * 2013-07-05 2015-12-23 (주)바이오니아 뎅기 바이러스 특이적 siRNA, 그러한 siRNA 를 포함하는 이중나선 올리고 RNA 구조체 및 이를 포함하는 뎅기 바이러스 증식 억제용 조성물
KR101715228B1 (ko) * 2013-07-05 2017-03-13 (주)바이오니아 뎅기 바이러스 특이적 siRNA, 그러한 siRNA 를 포함하는 이중나선 올리고 RNA 구조체 및 이를 포함하는 뎅기 바이러스 증식 억제용 조성물
CN105441588A (zh) * 2015-12-21 2016-03-30 深圳市生科源技术有限公司 登革热ⅰ、ⅱ、ⅲ、ⅳ型rt-pcr一步mix检测试剂盒及其检测方法
CN105441588B (zh) * 2015-12-21 2019-01-15 深圳生科原生物股份有限公司 登革热ⅰ、ⅱ、ⅲ、ⅳ型rt-pcr一步mix检测试剂盒及其检测方法
CN110760614A (zh) * 2019-04-29 2020-02-07 杭州市上城区疾病预防控制中心 一种登革热病毒核酸快速检测、分型以及溯源区分的方法
CN110760614B (zh) * 2019-04-29 2023-06-20 杭州市上城区疾病预防控制中心 一种登革热病毒核酸快速检测、分型以及溯源区分的方法

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