COMBINED ASSAY FOR HIV-1 AND HTLV-1 USING POLYMERASE CHA
COMBINED ASSAY FOR HIV-1 AND HTLV-1 USING POLYMERASE CHA
批准号:
3362207
负责人:
MARK M MANAK
金额:
$24.13万
依托单位国家:
美国
项目类别:
财政年份:
1989
资助国家:
美国
项目状态:
已结题
起止时间:
1989-08-15 至 1992-06-30
关键词:
AIDS /HIV test diagnosis design /evaluation enzyme linked immunosorbent assay globin human T cell lymphotropic virus type 1 human immunodeficiency virus 1 human subject lymphocyte molecular cloning natural gene amplification nucleic acid hybridization nucleic acid probes virus DNA virus classification
中文摘要
需要对感染逆转录病毒的细胞进行直接和灵敏的检测
用于确认患者暴露和监测疾病状态,以及
治疗。我们建议首先开发一种检测感染有病毒的PBL的方法
用聚合酶链式反应(PRC)扩增HTLV-I病毒
DNA和非放射性夹心杂交分析(在ELISA法中
微孔法)以检测扩增产物。这样的化验应该
能在10‘6个未感染的细胞中检测到一个感染的淋巴细胞。
初步工作将包括亚克隆、简化样品制备
以及HTLV-I和β-珠蛋白内对照试验的发展。
接下来是测试HTLV-I组合的可行性
用我们现有的HIV-1检测在聚合酶链式反应水平上进行检测以及
检测化验水平。组合测试应该会大大降低成本
使用DNA探针确认逆转录病毒暴露,节省时间和劳动力
和DNA样本。当选择最终的正式化验时,临床样本
将进行检测,并将HTLV-I/HIV-1 DNA结果与患者相关
抗体、临床和病毒分离数据。在后面的过程中,
根据这项提议,我们将为HIV-2和HTLV-II开发类似的检测方法。后
开发了单独的检测方法,两种检测方法的组合将是
经过测试并最终选定了格式。临床样本将进行测试,使用
联合化验正式,以确定敏感性和特异性
化验的结果。
英文摘要
Direct and sensitive assays for retrovirus infected cells are required
for confirming patient exposure and for monitoring disease status and
treatment. We propose first to develop an assay for PBLs infected with
HTLV-I by using the polymerase chain reaction (PRC) to amplify the viral
DNA and a nonradioactive sandwich hybridization assay (in an ELISA
microwell format) to detect the amplified product. Such an assay should
be capable of detecting one infected lymphocyte in 10'6 uninfected cells.
Initial work will entail subcloning, streamlining of sample preparation
and development of an HTLV-I and a beta-globin internal control assay.
This to be followed by testing the feasibility of combining the HTLV-I
assay with our existing HIV-1 assay at the PCR level as well as the
detection assay level. A combination test should greatly cut the cost of
confirming retroviral exposure, using DNA probes, by saving time, labor
and sample DNA. When the final assay formal is chose, clinical samples
will be tested and the HTLV-I/HIV-1 DNA results correlated with patient
antibody, clinical and viral isolation data. Later in the course of this
proposal, we will develop similar assays for HIV-2 and HTLV-II. After the
individual assays are developed, combinations of the two assays will be
tested and a final format chosen. Clinical samples will be tested, using
the combined assay formal, to determine the sensitivity and specificity
of the assay.,
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批准号:2064884
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资助金额:$26.62万
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财政年份:1990
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依托单位:
SIMPLE, RAPID PCR BASED ASSAY FOR LYME BORRELIOSIS
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批准号:2064885
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资助金额:$27.79万
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财政年份:1990
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DNA PROBES FOR THE DETECTION OF LYME DISEASE
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批准号:3489141
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资助金额:$4.91万
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财政年份:1990
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USE OF PCR FOR DIAGNOSING HIV-I INFECTION
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批准号:3489152
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项目类别:
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资助金额:$5.0万
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财政年份:1990
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负责人:MARK M MANAK
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依托单位:
USE OF PCR FOR DIAGNOSING HIV-1 INFECTION IN CHILDREN
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批准号:3506122
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项目类别:
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资助金额:$25.15万
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财政年份:1990
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负责人:MARK M MANAK
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依托单位:
USE OF PCR FOR DIAGNOSING HIV-1 INFECTION IN CHILDREN
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批准号:3506123
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项目类别:
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资助金额:$24.85万
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财政年份:1990
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负责人:MARK M MANAK
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依托单位:
COMBINED ASSAY FOR HIV-1 AND HTLV-1 USING POLYMERASE CHA
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批准号:2221082
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项目类别:
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资助金额:$22.83万
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财政年份:1989
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负责人:MARK M MANAK
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依托单位:
COMBINED ASSAY FOR HIV-1 AND HTLV-1 USING POLYMERASE CHA
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批准号:2221081
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项目类别:
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资助金额:$21.15万
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财政年份:1989
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负责人:MARK M MANAK
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依托单位:
NON-ISOTOPIC DNA PROBES TO DETECT HTLV-III NUCLEIC ACID
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批准号:3506421
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项目类别:
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资助金额:$19.11万
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财政年份:1986
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负责人:MARK M MANAK
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依托单位:
NON-ISOTOPIC DNA PROBES TO DETECT HTLV-III NUCLEIC ACID
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批准号:3506422
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项目类别:
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资助金额:$19.01万
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财政年份:1986
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负责人:MARK M MANAK
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依托单位:
DNA PROBE FOR DETECTION OF VIRAL NUCLEIC ACID
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批准号:3491070
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项目类别:
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资助金额:$3.59万
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财政年份:1985
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负责人:MARK M MANAK
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依托单位:
DNA TECHNIQUES FOR PRODUCTION OF A VACCINE TO AIDS
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批准号:3594774
-
项目类别:
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资助金额:$4.98万
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财政年份:1985
-
负责人:MARK M MANAK
-
依托单位:
DNA TECHNIQUES FOR PRODUCTION OF A VACCINE TO AIDS
-
批准号:3594775
-
项目类别:
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资助金额:$0.0万
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财政年份:1985
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负责人:MARK M MANAK
-
依托单位: