VIRAL GENE AMPLIFICATION ASSAYS FOR DETECTION OF CONTAMINATING MURINE VIRUSES
VIRAL GENE AMPLIFICATION ASSAYS FOR DETECTION OF CONTAMINATING MURINE VIRUSES
批准号:
6122943
负责人:
CYNTHIA L. BESCH-WILLIFORD
金额:
$0.0万
依托单位国家:
美国
项目类别:
财政年份:
1997
资助国家:
美国
项目状态:
已结题
起止时间:
1997-06-01 至 1999-05-31
关键词:
DNA virus Polyomavirus muris 1 RNA virus antiviral antibody communicable disease diagnosis cytomegalovirus diagnosis design /evaluation laboratory mouse laboratory rat lymphocytic choriomeningitis method development nucleic acid hybridization nucleic acid probes nucleic acid sequence polymerase chain reaction virus diseases virus genetics zoonosis
中文摘要
广泛使用源自或经过的生物材料
英文摘要
Extensive use of biologic materials originating from or passaged through
rodents for use in research investigations or human therapeutic regimes
has prompted increasing concern about detecting inadvertent
contamination of materials with rodent viruses. The diagnostic test
mouse frequently used to screen for virus contamination of these
biologic materials is the modified mouse or rat antibody production (MAP
or RAP) test, a bioassay involving a live animal component for antibody
production to viral contaminates, and a virus isolation component. The
time-consuming MAP/RAP test is plagues by variable sensitivities and
specificities. A more accurate and sensitive approach for the detection
of rodent viral contaminates in biologic specimens can be effected by
use of nucleic acid amplification and hybridization technologies. The
overall goal of this proposal is to develop gene amplification and
hybridization assays for the direct identification of rodent viruses in
biologic materials. The specific aims are as follows: 1.) Amplify gene
sequences by polymerase chain reaction (PCR) from each of a selected
list of murine RNA and DNA viruses known to contaminate biological
samples. The specificity of each assay evaluated by hybridization
experiments utilizing high-specificity, nonradioactive oligonucleotide
probes and nested primer sets. 2.) Develop complex PCR assays such that
multiple viruses can be detected in a single specimen by an
amplification assay containing primer paris specific for each virus. In
addition, liquid hybridization methods using enzyme-labelled
oligonucleotide probes will be developed to automate the amplicon
sequence confirmation procedures. 3.) Compare the specificity and
sensitivity of the PCR assays with that of the antibody production
tests. The sensitivity of the simple and complex PCR assays will be
calculated from cellular samples infected with known virus
concentrations, and compared with results of identical samples subjected
to the MAP/RAP test. Through the application of gene amplification and
hybridization technology, we will be capable of quickly and accurately
assessing murine virus contamination in reagents, and thus prevent the
use of these materials in human clinical treatments and in biological
research.
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TAS::75 0849::TAS 08024920111DA0
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批准号:8343276
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项目类别:
-
资助金额:$10.63万
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财政年份:2010
-
负责人:CYNTHIA L. BESCH-WILLIFORD
-
依托单位:
Operation of an Animal Diagnostic Laboratory
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批准号:7789039
-
项目类别:
-
资助金额:$10.0万
-
财政年份:2005
-
负责人:CYNTHIA L. BESCH-WILLIFORD
-
依托单位:
Operation of an Animal Diagnostic Laboratory
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批准号:8036362
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项目类别:
-
资助金额:$4.5万
-
财政年份:2005
-
负责人:CYNTHIA L. BESCH-WILLIFORD
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依托单位:
PATHOGENESIS OF MCMV INFECTION
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批准号:6319847
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项目类别:
-
资助金额:$0.13万
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财政年份:1999
-
负责人:CYNTHIA L. BESCH-WILLIFORD
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依托单位:--