US5290677A
Expired / Lapsed
US
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Rapid and sensitive test for detecting hepatitis A virus
Assignee: UNTIED STATES OF AMERICA HEALTH AND HUMAN SERVICES, Secretary of, Department of
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Filing: 1992-01-31
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Granted: 1994-03-01
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Expires: 2012-01-31
Abstract
Abstract Provided are methods for detecting hepatitis A virus (HAV) and HAV genotypes I and III by capturing whole hepatitis A virus with antibodies specific to hepatitis A virus, generating a cDNA copy of the RNA by reverse transcription in the presence of a negative strand primer having the sequence defined by SEQ ID NO:1, amplifying the cDNA by polymerase chain reaction with both the negative strand primer and a labelled or unlabelled positive strand primer having the sequence defined by SEQ ID NO: 2, and detecting the amplified cDNA by hybridization with either or both of the probes defined by SEQ ID NO:3 and SEQ ID NO:4 or by detection of label, wherein the presence of detectable hybridization or amplification indicates the presence of hepatitis A virus. Also provided are DNA primers and probes from the amino-terminal portion of the VPI region of the HAV genome that selectively hybridize with HAV, HAV genotype I and HAV genotype III and can amplify the amino-terminal portion of the VP1 region of HAV.
Inventors
- 👤 Betty H. Robertson
- 👤 Omana V. Nainan
- 👤 Vicki K. Brown
- 👤 Harold S. Margolis
- 👤 Bhawna Khanna
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-10-28 12:45
CPC Classifications
C; C12; C12Q; C12Q1/00; C12Q1/70; C12Q1/701; C12Q1/706; C12Q1/68; C12Q1/6804; C12Q1/6806; Y; Y02; Y02A; Y02A50/00; Y02A50/30
Claims Analysis
Total Claims: 14
Claim 1
Independent Claim
133 words
A method for detecting hepatitis A virus comprising the steps of: (i) capturing whole hepatitis A virus with antibodies specific to hepatitis A virus; (ii) generating a cDNA copy of the viral RNA by reverse transcription in the presence of a negative strand primer consisting of the nucleotides defined by SEQ ID NO:1; (iii) amplifying the cDNA of step (ii) by polymerase chain reaction with both said negative strand primer and a positive strand primer consisting of the nucleotides defined by SEQ ID NO: 2, said positive strand primer being labeled at the 5' end and wherein the amplification product can be used to distinguish hepatitis A genotypes I and III; and (iv) detecting the amplified cDNA of step (iii), wherein the presence of detectable amplification indicates the presence of hepatitis A virus.
Claim 2
Independent Claim
133 words
A method for detecting hepatitis A virus comprising the steps of: (i) capturing whole hepatitis A virus with antibodies specific to hepatitis A virus; (ii) generating a cDNA copy of the viral RNA by reverse transcription in the presence of a negative strand primer consisting of the nucleotides defined by SEQ ID NO:1; (iii) amplifying the cDNA of step (ii) by polymerase chain reaction with both said negative strand primer and a positive strand primer consisting of the nucleotides defined by SEQ ID NO: 2, said positive strand primer being labeled at the 5' end and wherein the amplification product can be used to distinguish hepatitis A genotypes I and III; and (iv) detecting the amplified cDNA of step (iii), wherein the presence of detectable amplification indicates the presence of hepatitis A virus.
Claim 3
Independent Claim
127 words
A method for detecting Genotype I of hepatitis A virus comprising the steps of: (i) capturing whole hepatitis A virus with antibodies specific to hepatitis A virus; (ii) generating a cDNA copy of the viral RNA by reverse transcription in the presence of a negative strand primer consisting of the nucleotides defined by SE ID NO:1; (iii) amplifying the cDNA of step (ii) by polymerase chain reaction with both said negative strand primer and a positive strand primer consisting of the nucleotides defined by SEQ ID NO: 2; and (iv) detecting the amplified cDNA of step (iii) by hybridization with the probe consisting of the nucleotides defined by SEQ ID NO:3, wherein the presence of detectable hybridization indicates the presence of Genotype I of hepatitis A virus.
Claim 4
Independent Claim
127 words
A method for detecting Genotype I of hepatitis A virus comprising the steps of: (i) capturing whole hepatitis A virus with antibodies specific to hepatitis A virus; (ii) generating a cDNA copy of the viral RNA by reverse transcription in the presence of a negative strand primer consisting of the nucleotides defined by SE ID NO:1; (iii) amplifying the cDNA of step (ii) by polymerase chain reaction with both said negative strand primer and a positive strand primer consisting of the nucleotides defined by SEQ ID NO: 2; and (iv) detecting the amplified cDNA of step (iii) by hybridization with the probe consisting of the nucleotides defined by SEQ ID NO:3, wherein the presence of detectable hybridization indicates the presence of Genotype I of hepatitis A virus.
Claim 5
Independent Claim
126 words
A method for detecting Genotype III of hepatitis A virus comprising the steps of: (i) capturing whole hepatitis A virus with antibodies specific to hepatitis A virus; (ii) generating a cDNA copy of the viral RNA by reverse transcription in the presence of a negative strand primer consisting of the nucleotides defined by SEQ ID NO:1; (iii) amplifying the cDNA of step (ii) by polymerase chain reaction with both said negative strand primer and a positive strand primer consisting of the nucleotides defined by SEQ ID NO:2; and (iv) detecting the amplified cDNA of step (iii) by hybridization with the probe consisting of the nucleotides defined by SEQ ID NO:4, wherein the presence of detectable hybridization indicates the presence of Genotype III of hepatitis A virus.
Claim 6
Independent Claim
126 words
A method for detecting Genotype III of hepatitis A virus comprising the steps of: (i) capturing whole hepatitis A virus with antibodies specific to hepatitis A virus; (ii) generating a cDNA copy of the viral RNA by reverse transcription in the presence of a negative strand primer consisting of the nucleotides defined by SEQ ID NO:1; (iii) amplifying the cDNA of step (ii) by polymerase chain reaction with both said negative strand primer and a positive strand primer consisting of the nucleotides defined by SEQ ID NO:2; and (iv) detecting the amplified cDNA of step (iii) by hybridization with the probe consisting of the nucleotides defined by SEQ ID NO:4, wherein the presence of detectable hybridization indicates the presence of Genotype III of hepatitis A virus.
Claim 7
Dependent Claim
18 words
A DNA primer that hybridizes with hepatitis A virus, consisting of the sequence defined by SEQ ID NO:1.
Claim 8
Dependent Claim
18 words
A DNA primer that hybridizes with hepatitis A virus, consisting of the sequence defined by SEQ ID NO:1.
Claim 9
Dependent Claim
18 words
DNA primer that hybridizes with hepatitis A virus, consisting of the sequence defined by SEQ ID NO: 2.
Claim 10
Dependent Claim
18 words
DNA primer that hybridizes with hepatitis A virus, consisting of the sequence defined by SEQ ID NO: 2.
Claim 11
Dependent Claim
20 words
A DNA probe that hybridizes with hepatitis A virus genotype I, consisting of the sequence defined by SEQ ID NO:3.
Claim 12
Dependent Claim
20 words
A DNA probe that hybridizes with hepatitis A virus genotype I, consisting of the sequence defined by SEQ ID NO:3.
Claim 13
Dependent Claim
20 words
A DNA probe that hybridizes with hepatitis A virus genotype III, consisting of the sequence defined by SEQ ID NO:4.
Claim 14
Dependent Claim
20 words
A DNA probe that hybridizes with hepatitis A virus genotype III, consisting of the sequence defined by SEQ ID NO:4.
References Cited (Backward Citations)
US5075212A
Cited
USPTO Annuity & Maintenance Windows
| Annuity Stage | Window Opens | Payment Deadline | Surcharge Period Until | Estimated Fee | Status |
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| 3.5 Years (Year 4) | 1997-02-28 | 1997-08-30 | 1998-03-01 | $2000.00 | Window Closed / Lapsed |
| 7.5 Years (Year 8) | 2001-02-28 | 2001-08-30 | 2002-03-01 | $3760.00 | Window Closed / Lapsed |
| 11.5 Years (Year 12) | 2005-02-28 | 2005-08-30 | 2006-03-01 | $7700.00 | Window Closed / Lapsed |