WO1991014789A1 - Detection d'une infection par hepatite - Google Patents

Detection d'une infection par hepatite Download PDF

Info

Publication number
WO1991014789A1
WO1991014789A1 PCT/GB1991/000503 GB9100503W WO9114789A1 WO 1991014789 A1 WO1991014789 A1 WO 1991014789A1 GB 9100503 W GB9100503 W GB 9100503W WO 9114789 A1 WO9114789 A1 WO 9114789A1
Authority
WO
WIPO (PCT)
Prior art keywords
hbv
hepatitis
variant
absence
patients
Prior art date
Application number
PCT/GB1991/000503
Other languages
English (en)
Inventor
Howard Christopher Thomas
William Frederick Carman
Original Assignee
Imperial College Of Science, Technology & Medicine
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 Imperial College Of Science, Technology & Medicine filed Critical Imperial College Of Science, Technology & Medicine
Publication of WO1991014789A1 publication Critical patent/WO1991014789A1/fr

Links

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
    • C12Q1/68—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
    • C12Q1/6858—Allele-specific amplification
    • 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

Definitions

  • This invention relates to a method to directly detect a pre-core variant of Hepatitis B virus (HBV) in sera and its application to predict outcome in patients with fulminant or chronic HBV infection.
  • HBV Hepatitis B virus
  • Carmen at al (The Lancet 1989; ii: 588-591) have described a pre-core variant of HBV having a mutation from guanosine to adenosine at position 1896 from the unique Eco Rl site and a variable mutation from guanosine to adenosine may may or may not be present at position 1899. These mutations are referred to hereinafter as mu-1896 and mu-1899 respectively. Sequencing of sera from patients with acute and chronic Hepatitis suggests that the pre-core variant is a common occurrence in the life-cycle of the virus, occurring at, or shortly before, the seroconversion to anti-HBe.
  • mu-1896 essentially results in the production of a translational stop codon and resulting inability to produce HBe antigen HBeAg) .
  • the mutation which is unable to produce HBeAg is the "HBe Antigen -ve "virus (hereinafter named the e-ve virus) whilst the virus which can produce HBeAg is the "HBe antigen +ve “virus (hereinafter named the e +ve virus) .
  • viruses As patients come nearer to seroconversion to anti-HBe, they are more likely to respond to antiviral therapy with synthetic or natural antiviral compounds, including natural and recombinant alpha, beta and gamma interferons. Although some factors predictive of response have been identified by Brook et al, Hepatology 1990; 10; 761-763, including low serum HBV-DNA levels and high serum transaminase concentrations, other more
  • SUBSTITUTESHEET precise predictive factors are required so that only those patients with a high probability of response are treated with the expensive and relatively toxic currently available antiviral compounds.
  • the present invention provides a process for aiding prediction of the outcome of Hepatitis B infection characterised by testing for the presence or absence of the above-mentioned pre-core variant of HBV in a sample from a patient.
  • the invention also provides the use in prediction of the outcome of Hepatitis B infection of a test for the presence or absence of the above-mentioned pre-core variant of HBV in a sample from a patient.
  • pre-core variant we mean a variant having mu-1896 with or without mu-1899.
  • a modification of the polymerase chain reaction is used to detect the variant, without the necessity of sequencing, by means of an appropriate combination of oligonucleotide primers which recognise the target mutations.
  • a common primer is used to the pre-core region of HBV in combination with one of a pair of discriminatory primers in each of two reaction vessels.
  • the method for extraction of HBV-DNA from serum is as detailed in Carmen et al.
  • the Lancet 1989, ii : 588-591 and the method for PCR is also as detailed with the exception that the annealing temperature is 60°C for 2 minutes.
  • the discrimination is based on a single base substitution at the 3-prime end of the primer, such that the one primer will bind to the mutation but not the other (which will bind to the normal strain) , allowing either DNA of the mutant or the normal virus, respectively, to be amplified. Further details.are illustrated in Figure 1 which sets out partial nucleotide sequences for selected primers.
  • first, second and third variable (discriminatory) primers 11, 12 and 13 bind to the negative strand 15 of HBV-DNA of normal strain.
  • the other end of the PCR product is defined by the binding site of common primer 14 to the positive strand.
  • 3' terminal nucleotides at positions 1896 are indicated for the variable primers and positions 1896 and 1899 are shown for variable primer 11.
  • primer 11 has mu-1896 as well as mu-1899.
  • the PCR products will be dotted onto a filter using standard technology and probed with a labelled, radioactively or non radioactively, HBV-DNA.
  • a further technology is to add ethidium bromide to the PCR products, add this to a filter which is able to bind double-stranded but not a single-stranded DNA, wash the filter and examine under ultra-violet light for fluorescence.
  • a chronic infection with an e +ve virus With a chronic infection with an e +ve virus the patient may recover without treatment (about 10% of cases) . When treatment with antiviral therapy is necessary it is usually of short duration (about 3 months) .
  • a chronic infection with an e -ve virus is more rapidly progressive and usually requires antiviral therapy over many months, ie lengthy maintenance treatment. In the longer term different antiviral treatments may be used to treat e -ve and e tve infections.
  • An e -ve infection carries a greater risk of transmission of a fulminant infection to a sexual partner or to an offspring.
  • An e +ve infection generally transmits a chronic infection to an offspring and an acute non-fulminant infection to a sexual partner,
  • kits containing the essential components required ie to . assay kits for diagnostic, monitoring and/or testing purposes which comprise means for determining the presence or absence of a pre-core variant of HBV having mu-1896 with or without mu-1899 together with appropriate packaging and instructions for performing a process as described in this specification according to any feature of the present invention.
  • assay kits for diagnostic, monitoring and/or testing purposes which comprise means for determining the presence or absence of a pre-core variant of HBV having mu-1896 with or without mu-1899 together with appropriate packaging and instructions for performing a process as described in this specification according to any feature of the present invention.
  • kits include for example:
  • a container eg a tube containing primer 14 and 13, another with 14 and 12 and another with 14 and 11:
  • a container eg a tube
  • eg 5 x amplification buffer containing a mixture of deoxynucleotides
  • a body fluid sample is taken from a patient suffering from hepatitis and is treated, for example with PCR, to ascertain if the infection is caused by an e -ve or an e +ve virus and thereafter appropriate treatment is recommended depending upon the result.
  • the invention is illustrated by the following example:
  • This example relates to the sequencing of the pre-core and core regions of HBV DNA contracted from serum from 18 patients with fulminant acute hepatitis B.
  • hepatitis B- virus has been described recently in hepatitis B surface antigen positive Mediterranean patients who lack hepatitis B e antigen and who have an unusual and severe form of chronic hepatitis. This variant is unable to produce hepatitis B e antigen because of the presence of a novel translational stop codon at the end of the precore region of the genome.
  • Fulminant hepatic failure was defined according to the criteria of Trey and Davidson ("Progress in Liver Disease", New York, Greene & Stralton, 1970, 282-298) (encephalopathy within 8 weeks of first symptoms) .
  • Sera were obtained from patients seen at two referral institutions. Subjects were chosen from consecutive admissions on the basis that sera and clinical details were available from the date of initial admission. Twelve patients (nos. 1-12) were seen at Kings College Hospital, London, United Kingdom. 11 had fulminant hepatitis of who 6 were HBeAg positive, 3 anti-HBe positive and 2 had neither "e” marker on admission, although one of the latter developed anti-HBe 2 days later. DNA was amplified from 8 of the 11 patients and sequence data obtained.
  • Patients 11 and 12 were sisters who were intravenous drug abusers and had been infected from a shared needle; one (patient 11) had a fulminant course and the other developed acute benign disease with HBeAg in serum. Sequence data were obtained from the latter only.
  • Sera were tested for HBsAG, HBeAG, IgM anti-HBe and anti-HBe using assays from Abbott Laboratories, UK. Hepatitis delta antigen and antibody were measured using i-mmunoassays from Ricktech, UK. Sera 1 to 8 were tested for antibody to hepatitis C virus using an Ortho Diagnostics assay. Serum HBV-DNS was assayed by molecular hybridisation (J.Med. Virology, 1982; jL, 273-280) . Extraction of DNS and PCR-sequencing Serum (50ul) ws processed as described in The Lancet, 1989; ii, 588-591. Primers IVTC 1
  • SUBSTITUTESHEET hepatitis patients all were anti-HBe position, 4 died and one underwent liver transplantation. They presented at an average of 7 days (range 3 to 10) after onset of symptoms, which was similar to that of anti-HBe positive patients at the first centre. The four patients died 2 to 3 days after admission, strikingly earlier than the HBeAG positive patients at the first centre. Of the 2 HBeAG positive patients with hepatitis delta virus infection, one recovered and the other was transplanted. PCR results
  • Sequencing results could be obtained from 15 of the fulminant cases and all 3 of the suspected contacts with hepatitis B (table) .

Landscapes

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

Abstract

Détection de l'hépatite B au moyen d'un procédé dans lequel on effectue un test pour vérifier la présence ou l'absence d'une variante de pré-noyau du virus de l'hépatite B (HBV) comportant des mutations se produisant à la position 1896 (mu-1896) avec ou sans mutation ultérieure se produisant à la position 1899 (mu-1899) dans un échantillon prélevé sur un malade. mu-1896 aboutit essentiellement à la production d'un codon d'arrêt de translation et de l'incapacité qui en résulte de produire un antigène HBe. Le procédé, le kit d'analyse ainsi que la mise en application du procédé font tous partie des revendications.
PCT/GB1991/000503 1990-03-28 1991-03-28 Detection d'une infection par hepatite WO1991014789A1 (fr)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
GB9006924.6 1990-03-28
GB909006924A GB9006924D0 (en) 1990-03-28 1990-03-28 Prognosis of hepatitis infection

Publications (1)

Publication Number Publication Date
WO1991014789A1 true WO1991014789A1 (fr) 1991-10-03

Family

ID=10673398

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/GB1991/000503 WO1991014789A1 (fr) 1990-03-28 1991-03-28 Detection d'une infection par hepatite

Country Status (2)

Country Link
GB (1) GB9006924D0 (fr)
WO (1) WO1991014789A1 (fr)

Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP0593789A1 (fr) * 1992-05-08 1994-04-27 Sumitomo Metal Industries, Ltd. Methode de determination de la mutation pre-c du virus de l'hepatite b
US8642752B2 (en) 2011-04-21 2014-02-04 Isis Pharmaceuticals, Inc. Modulation of Hepatitis B virus (HBV) expression

Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP0317239A2 (fr) * 1987-11-13 1989-05-24 Native Plants Incorporated Procédé et dispositif pour la détection des polymorphismes de restriction des longueurs de fragments
EP0333465A2 (fr) * 1988-03-18 1989-09-20 Baylor College Of Medicine Détection de mutations utilisant des oligonucléotides d'amorce compétitives
EP0348529A1 (fr) * 1987-12-25 1990-01-03 Wakunaga Seiyaku Kabushiki Kaisha Procede de detection d'un acide nucleique cible dans un specimen
EP0332435B1 (fr) * 1988-03-10 1992-04-22 Zeneca Limited Procédé pour détecter des séquences de nucléotides

Patent Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP0317239A2 (fr) * 1987-11-13 1989-05-24 Native Plants Incorporated Procédé et dispositif pour la détection des polymorphismes de restriction des longueurs de fragments
EP0348529A1 (fr) * 1987-12-25 1990-01-03 Wakunaga Seiyaku Kabushiki Kaisha Procede de detection d'un acide nucleique cible dans un specimen
EP0332435B1 (fr) * 1988-03-10 1992-04-22 Zeneca Limited Procédé pour détecter des séquences de nucléotides
EP0333465A2 (fr) * 1988-03-18 1989-09-20 Baylor College Of Medicine Détection de mutations utilisant des oligonucléotides d'amorce compétitives

Non-Patent Citations (4)

* Cited by examiner, † Cited by third party
Title
Biological Abstracts, volume 88, no. 11, December 1989, M.R. Brunetto et al.: "Identification of HBV variants which cannot produce precore derived HBeAg and may be responsible for severe hepatitis", see page 404 *
Chemical Abstracts, volume 112, no. 23, June 1990, (Columbus, Ohio, US), Okamoto, Hiroaki et al.: "Hepatitis B viruses with precore region defects prevail in persistently infected hosts along with seroconversion to the antibody against e antigen", see page 138 *
Proc. Natl. Acad. Sci., volume 86, December 1989, Chemistry, (US), F.F. Chebab et al.: "Detection of specific DNA sequences by fluorescense amplification: A color complementation assay", pages 9178-9182 *
The Lancet, volume II, 9 September 1989, W.F. Carman et al.: "Mutation preventing formation of hepatitis B e antigen in patients with chronic hepatitis B infection"+, pages 588-591 *

Cited By (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP0593789A1 (fr) * 1992-05-08 1994-04-27 Sumitomo Metal Industries, Ltd. Methode de determination de la mutation pre-c du virus de l'hepatite b
EP0593789A4 (en) * 1992-05-08 1998-02-04 Sumitomo Metal Ind Method of judging pre-c mutation of hepatitis b virus
US8642752B2 (en) 2011-04-21 2014-02-04 Isis Pharmaceuticals, Inc. Modulation of Hepatitis B virus (HBV) expression
US9034841B2 (en) 2011-04-21 2015-05-19 Isis Pharmaceuticals, Inc. Modulation of hepatitis B virus (HBV) expression
US9127278B2 (en) 2011-04-21 2015-09-08 Isis Pharmaceuticals, Inc. Modulation of hepatitis B virus (HBV) expression
US9677076B2 (en) 2011-04-21 2017-06-13 Ionis Pharmaceuticals, Inc. Modulation of hepatitis B virus (HBV) expression

Also Published As

Publication number Publication date
GB9006924D0 (en) 1990-05-23

Similar Documents

Publication Publication Date Title
Handa et al. Prevalence of the newly described human circovirus, TTV, in United States blood donors
Roth et al. NAT for HBV and anti‐HBc testing increase blood safety
Hyypiä et al. Detection of enteroviruses by spot hybridization
Nolte et al. Preclinical evaluation of AMPLICOR hepatitis C virus test for detection of hepatitis C virus RNA
Nakatsuji et al. Prevalence and disease association of hepatitis G virus infection in Japan
EP0532258A2 (fr) Oligonucléotides et procédé pour la détection des génotypes de HCV
NO329925B1 (no) Fremgangsmate for a detektere og amplifisere HCV-RNA, oligonukleotid, primer, probe og kit.
Lefrère et al. Prevalence of GB virus type C/hepatitis G virus RNA and of anti‐E2 in individuals at high or low risk for blood‐borne or sexually transmitted viruses: evidence of sexual and parenteral transmission
Laskus et al. Exposure of hepatitis C virus (HCV) RNA-positive recipients to HCV RNA-positive blood donors results in rapid predominance of a single donor strain and exclusion and/or suppression of the recipient strain
Pao et al. Serum hepatitis B virus DNA in hepatitis B virus seropositive and seronegative patients with normal liver function
Heo et al. Detection of YMDD motif mutants by oligonucleotide chips in lamivudine-untreated patients with chronic hepatitis B virus infection
Zöllner et al. Correlation of hepatitis B virus load with loss of e antigen and emerging drug‐resistant variants during lamivudine therapy
JPH09501829A (ja) 肝炎ｂ型ウイルスのヌクレオチド配列及びｄｎａの増幅及び検出のための方法
KR100910545B1 (ko) Ｃ형 간염 바이러스(ｈｃｖ) 및 사람 면역결핍 바이러스(ｈｉｖ)의 효과적인 다중 검출을 위한 올리고뉴클레오티드 프라이머, 이의 이용 방법, 및 이를 포함하는 키트
Zhong et al. Quantitative and genotypic analysis of TT virusinfection in Chinese blood donors
Venegas et al. Prevalence of hepatitis B virus genotypes in chronic carriers in Santiago, Chile
JP4237823B2 (ja) 肝炎ｇｂウイルスの核酸検出
WO1991014789A1 (fr) Detection d'une infection par hepatite
Ferraz et al. Fulminant hepatitis in patients undergoing liver transplantation: Evidence for a non‐A, non‐B, non‐C, non‐D, and non‐E syndrome
Enan et al. Analysis of hepatitis B virus-mixed genotype infection by ultra deep pyrosequencing in Sudanese patients, 2015–2016
Roque-Afonso et al. Monitoring the Emergence of Hepatitis B Virus Polymerase Gene Variants during Lamivudine Therapy in Human Immunodeficiency Virus Coinfected Patients: Performance of Clip™ Sequencing and Line Probe Assay
Wang et al. Simultaneous detection of HBV and HCV by multiplex PCR normalization
JP2010246555A (ja) Ｃ型肝炎ウイルスの検出及び定量方法
AHMED et al. Hepatitis C viral load, genotype and histological severity in patients with bleeding disorders
Chakravarti et al. Distribution of hepatitis C virus genotypes in beta‐thalassaemic patients from Northern India

Legal Events

Date Code Title Description
AK Designated states

Kind code of ref document: A1

Designated state(s): CA JP US

AL Designated countries for regional patents

Kind code of ref document: A1

Designated state(s): AT BE CH DE DK ES FR GB GR IT LU NL SE

NENP Non-entry into the national phase

Ref country code: CA