Chemistry: molecular biology and microbiology – Measuring or testing process involving enzymes or... – Involving virus or bacteriophage
Patent
1992-06-26
1997-04-15
Jones, W. Gary
Chemistry: molecular biology and microbiology
Measuring or testing process involving enzymes or...
Involving virus or bacteriophage
4352523, 4353201, 436536, 5303883, 5303894, C12Q 170
Patent
active
056208430
DESCRIPTION:
BRIEF SUMMARY
This application is the national filing of PCT application No. PCT/EP91/01110.
The invention relates to a nucleic acid sequence coding for a peptide or a fragment thereof which is immunochemically reactive with NANBH virus antibodies (Non-A Non-B Hepatitis).
The invention also relates to a method for the detection of NANBH or anti-NANBH in a test fluid and also to an immunochemical reagent and a test kit for carrying out the said detection methods.
Non-A, Non-B hepatitis which may or may not be caused by Hepatitis C Virus (HCV) is a transmissible disease or family of diseases shown to be virus-induced. It can be distinguished from other forms of viral-associated liver diseases, including that caused by the known hepatitis viruses, i.e., hepatitis A virus (HAV), hepatitis B virus (HBV), and delta hepatitis virus (HDV), as well as the hepatitis induced by cytomegalovirus (CMV) or Epstein-Barr virus (EBV). NANBH was first identified in transfused individuals. Transmission from man to chimpanzee and serial passage in chimpanzees provided evidence that NANBH is due to a transmissible infectious agent or agents.
Epidemiologic evidence is suggestive that three types of NANBH exist: the water-borne epidemic type; the blood or needle associated type; and the sporadically occurring (community acquired) type. However, the number of agents which may be the causative of NANBH is unknown.
Clinical diagnosis and identification of NANBH has been accomplished primarily by exclusion of other viral markers. Among the methods used to detect putative NANBH antigens and antibodies are agar-gel diffusion, counter-immunoelectrophoresis, immunofluorescence microscopy, immune electron microscopy, radioimmunoassay, and enzyme-linked immunosorbent assay. However, none of these assays has proved to be sufficiently sensitive, specific, and reproducible to be used as a diagnostic test for NANBH.
However, for the development of a specific and sensitive method to enable a reliable diagnosis to be made in various phases of the infection with NANBH it is of great importance to identify immuno-dominant viral epitopes of this type.
A nucleic acid sequence according to FIG. 1 (SEQ. ID NO: 5) or a fragment thereof has now been found coding for a peptide which is surprisingly immunochemically reactive with NANBH-antibodies.
The invention further comprises a peptide with 87 amino acids and an amino acid sequence as shown in FIG. 2 which is immunochemically reactive with NANBH antibodies. Said amino acid sequence does not correspond to any published amino acid sequence with regard to NANBH (for example the published sequence of Chiron in EP 318,216).
The invention also comprises fragments of the said peptide which are still immunochemically reactive with NANBH-antibodies and also polypeptides comprising the said peptide or said fragment thereof.
The invention also relates to an immunochemical reagent, which reagent comprises at least one of the peptides or fragments thereof according to the invention.
The invention also comprises a method for the detection of antibodies directed against NANBH in a test fluid, using at least one of the peptides according to the invention.
The invention also relates to a method for the detection of NANBH in a test fluid, using at least one of the peptides according to the invention.
The invention also relates to a test kit to be used in an immuno-assay, said test kit containing at least an immunochemical reagent according to the invention.
It is within the scope of this invention to use the new nucleotide sequence according to FIG. 1 as the basis of a test to detect NANBH DNA or RNA by a nucleic acid amplification technique for instance the polymerase chain reaction (PCR) or the nucleic acid sequence based amplification (NASBA), as described in EP 201,184 and EP 329,822, respectively.
Moreover, a peptide or fragment thereof according to the invention can be used in suitable pharmaceutical dosage forms in the treatment of NANB Hepatitis-disease. The preparation of vaccines thus obtained which contain a peptide or fra
REFERENCES:
The 1988/89 Promega Catalog, Chapter 12, p. 10.
K. Takeuchi et al., "Hepatitis C viral cDNA clones isolated from a healthy carrier donor implicated in post-transfusion non-A, non-B hepatitis," Gene, vol. 91, No. 2, pp. 287-291, Jul. 16, 1990, The Netherlands.
H. Okamoto et al., "The 5'-terminal sequence of the hepatitis C virus genome," Japan. J. Exp. Med., vol. 60, No. 3, pp. 166-177, 1990.
Y. Kubo et al., "A cDNA fragment of hepatitis C virus isolated from an implicated donor of post-transfusion non-A, non-B hepatitis in Japan," Nucleic Acid Research, vol. 17, No. 24, pp. 10367-10372, 1989.
Hellings Jan A.
Wilhelmus de Haard Johannes J.
Akzo Nobel N.V.
Blackstone William M.
Gormley Mary E.
Houtteman Scott
Jones W. Gary
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