Chemistry: molecular biology and microbiology – Measuring or testing process involving enzymes or... – Involving virus or bacteriophage
Patent
1992-05-22
1994-12-20
Saunders, David
Chemistry: molecular biology and microbiology
Measuring or testing process involving enzymes or...
Involving virus or bacteriophage
435 795, 435975, 436812, 530325, 530326, 530329, C07K 706, C07K 708, C07K 710, G01N 33569
Patent
active
053745175
DESCRIPTION:
BRIEF SUMMARY
TECHNICAL FIELD
THIS INVENTION is directed to the diagnosis and treatment of herpes virus related diseases. In particular, it relates to the use of specific open reading frames (ORFs) within the Epstein-Barr virus (EBV) which encode antigens recognized by EBV-specific antibodies raised during infectious mononucleosis (IM) and related diseases and the use of synthetic peptides based on the amino acid sequences encoded by these ORFs in a specific and reliable diagnostic test for, and treatment of, IM and related diseases.
BACKGROUND ART
Epstein-Barr virus (EBV) is a member of the herpes virus family and is present in all human populations. Primary infection usually occurs in early childhood and remains silent throughout a person's life. However, when uninfected adolescents and young adults are exposed to EBV, about 60% manifest infectious mononucleosis (IM).
The predominant laboratory test used to establish the diagnosis of IM has been the demonstration of heterophil antibodies. The rapid slide tests have become the most widely used method to detect these heterophil antibodies. In contrast, quantitative agglutination tests, such as the Paul-Bunnell-Davidsohn method, are more accurate but are also more tedious and time consuming.
The use of tests to measure heterophil antibodies have limitations, namely, only between 80-95% of IM patients produce these antibodies, these antibodies are absent in a large percentage of young children and the antibodies are produced in a variety of other diseases such as lymphoma, hepatitis and leukemia. The measurement of heterophil antibodies also does not give any indication of the severity of the disease and cannot be used to monitor the course of IM.
Immunofluoresence tests that measure antibodies to EBV can also be used in the diagnosis of heterophil-negative cases of IM, or patients with atypical manifestations.
These tests, however, are time consuming and require the use of trained personnel and specialized equipment which does not make them amenable to the routine analysis of large numbers of samples.
The diagnosis of an acute primary EBV infection can also be determined by an IgM response to EBV-viral capsid antigen (VCA). The VCA is composed of a large number of different antigens. Components of VCA are defined by the fact that they are expressed late in the replicative cycle of the virus. Many VCA components have been mapped to specific open reading frames (ORF's) within the EBV genome though there are many ORF's, known to be expressed late in replication, to which specific VCA antigens have not yet been identified.
Genetic engineering and synthetic polypeptide technologies now enable the manufacture of large quantities of protein and polypeptide antigens. However, these techniques are only effective if the amino acid residue sequence of the native protein is known.
The amino acid residue sequence of a natural protein can be determined by sequencing of the protein itself.
Alternatively, the DNA sequence that codes for the protein may also reveal the protein's amino acid residue sequence.
Antibodies can be used to determine whether an ORF present in a DNA sequence codes for a protein. This involves manufacturing an array of protein fragments or synthetic polypeptides whose amino acid residue sequences correspond to the hypothetical sequences obtained from the ORFs. The protein fragments or polypeptides to which naturally occurring antibodies immunoreact thereby identify the ORF as encoding a naturally occurring protein. The complete amino acid sequence of this protein could then be deduced from the DNA sequence of the ORF.
DISCLOSURE OF THE INVENTION
It is a general object of the present invention overcome, or at least ameliorate, one or more of the above disadvantages, and to provide a specific and reliable test for the diagnosis for, and treatment of, IM and related diseases.
As the complete DNA sequence for EBV has been identified, including the start and stop codons which, prima facie, define potential ORFs for the transcription of the genetic code, the pres
REFERENCES:
patent: 4554101 (1985-11-01), Hopp
patent: 4707358 (1987-11-01), Kieff et al.
Middeldorp et al., 1988b. Epitope mapping on the Epstein-Barr virus major capoid protein using systemic synthesis of overlapping oligopeptides. J. Virolog. Meth. 21:147-59.
Middeldorp et al., 1988a. Epstein-Barr virus specific marker molecules for early diagnosis of infectious mononucleosis, J. Virolog. Meth., 21:133-46.
Sculley et al., 1986. Reactions of sera from patients with rheumatord arthritis systemic lupus erythematosus and infections mononucleosis to Epstein-Barr Virus-induced Polypeptides, J. Gen. Virol., 67:2253-8.
Sculley et al., 1985. Identification of Epstein-Barr Virus-induced polypeptides in P3HR-1 cells by protein immunoblot. J. Gen. Virol. 66:1113-22.
Baer et al., 1984. DNA sequence and expression of the B95-8 Epstein-Barr virus genome. Nature 310:207-11.
Mackett et al., Jun. 1990, Characterization and expression of a glycoprotein encoded by the Epstein-Barr virus Bam HI I fragment. J. Virol. 64:2545-52.
Seguin et al., 1983. DNA sequence and transcription of the Bam HI fragment B region of B95-8 Epstein-Barr virus. Mol. Biol. Med., 1:369-92.
Parker et al., 1986. New hydrophilicity seale derived from high-performance liquid chromatography peptide retention data: correlation of predicted surface residue with antigenicity and X-ray-derived accessible sites. Biochem. 25:5425-32.
Kinney et al., 1989. The full-length nucleotide sequence of the verulent Trinidad Donkey Strain of Venezualan equine encephalitis virus and its attenuated vaccine derivative, strain TC-83. Virol. 170:19-30.
Walls et al.; "The analysis of EBV proteins which are antigenic in vivo"; Nucleic Acids Research, vol. 16, No. 7, 1988.
Moss Denis J.
Sculley Tom B.
Council of the Queensland Institute of Medical Research
Grun James L.
Saunders David
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