Title:
【発明の名称】ノーワーク及び関連のウィルスを検出し同定するための方法と試薬
Document Type and Number:
Japanese Patent JP3310281
Kind Code:
B2
Abstract:
Double-stranded cDNA was synthesized from nucleic acid extracted from Norwalk virus purified from stool specimens of volunteers. One clone was isolated from a cDNA library constructed in a pUC-13 vector after amplification of the cDNA. The specificity of this cDNA (pUCNV-953) was shown by hybridization assays. The cDNA reacted with post- (but not pre-) infection stool samples from Norwalk volunteers and with highly purified Norwalk virus, but not with other common enteric viruses such as hepatitis A virus and rotavirus. Finally, the probe detected virus in the same fractions of CsCl gradients in which viral antigen was detected using a specific Norwalk virus radioimmunoassay, and particles were detected by immune electron microscopy. Single-stranded RNA probes derived from the DNA clone after subcloning into an in vitro transcription vector were also used to show that the Norwalk virus contains a ssRNA genome of about 8 kb in size. The original clone was also used to detect additional cDNAs which represent at least 7 kb of nucleic acid of the Norwalk genome. The availability of a Norwalk-specific cDNA and the first partial genome sequence information allow rapid cloning of the entire genome and of establishment of sensitive diagnostic assays. Such assays can be based on detection of Norwalk virus nucleic acid or Norwalk viral antigen using polyclonal or monoclonal antibodies to proteins expressed from the cDNA or to synthetic peptides made based on the knowledge of the genome sequence. Vaccines made by recombinant DNA technology are now feasible.
Inventors:
Estas, Mary Kay.
Jean, J
Graham, David Wye.
Jean, J
Graham, David Wye.
Application Number:
JP51569690A
Publication Date:
August 05, 2002
Filing Date:
October 30, 1990
Export Citation:
Assignee:
Baylor College of Medicine
International Classes:
C07K14/08; C07K16/10; C12N9/12; C12N15/02; C12N15/09; C12N15/33; C12P19/34; C12N5/10; C12P21/08; C12Q1/70; G01N33/53; G01N33/569; G01N33/577; C12R1/91; C12R1/92; (IPC1-7): C12N15/09; C12N5/10; C12N9/12; G01N33/53; G01N33/569; G01N33/577
Attorney, Agent or Firm:
Morio Sada