利用聚合酶链反应和基于时间分辨荧光的杂交分析法检测1型人类免疫缺陷病毒
Detection of human immunodeficiency virus type 1 by using the polymerase chain reaction and a time-resolved fluorescence-based hybridization assay.
作者信息
Dahlén P O, Iitiä A J, Skagius G, Frostell A, Nunn M F, Kwiatkowski M
机构信息
Pharmacia Genetic Engineering Inc., La Jolla, California 92037.
出版信息
J Clin Microbiol. 1991 Apr;29(4):798-804. doi: 10.1128/jcm.29.4.798-804.1991.
Abstract
The polymerase chain reaction (PCR) has many potential applications in the field of nucleic acid diagnostics. In particular, it has been successfully applied to the detection of pathogens present in low copy numbers such as the human immunodeficiency virus type 1. Here we describe a time-resolved fluorescence-based hybridization assay which, combined with the PCR, offers an extremely sensitive method for the detection of nucleic acids. In this assay format, the PCR is run by standard procedures and the subsequent hybridization reaction is carried out in solution by using two oligonucleotide probes, one biotinylated and one labeled with europium (Eu3+). The sandwich hybrids are then collected onto a streptavidin-coated microtitration well, and the bound Eu3+ is measured in a time-resolved fluorometer. This assay is rapid, user friendly, and quantitative and lends itself to automation. The application of this assay to the detection of human immunodeficiency virus type 1 is described.
摘要
聚合酶链反应（PCR）在核酸诊断领域有许多潜在应用。特别是，它已成功应用于检测低拷贝数存在的病原体，如1型人类免疫缺陷病毒。在此，我们描述一种基于时间分辨荧光的杂交检测方法，该方法与PCR相结合，为核酸检测提供了一种极其灵敏的方法。在这种检测形式中，PCR按标准程序进行，随后的杂交反应在溶液中通过使用两种寡核苷酸探针进行，一种用生物素标记，另一种用铕（Eu3+）标记。然后将夹心杂交体收集到包被链霉抗生物素蛋白的微量滴定孔上，并在时间分辨荧光计中测量结合的Eu3+。该检测方法快速、用户友好且定量，适合自动化操作。本文描述了该检测方法在1型人类免疫缺陷病毒检测中的应用。
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本文引用的文献
1
Rapid prenatal diagnosis of sickle cell anemia by a new method of DNA analysis.通过一种新的DNA分析方法对镰状细胞贫血进行快速产前诊断。
N Engl J Med. 1987 Mar 12;316(11):656-61. doi: 10.1056/NEJM198703123161103.
2
Analysis of enzymatically amplified beta-globin and HLA-DQ alpha DNA with allele-specific oligonucleotide probes.用等位基因特异性寡核苷酸探针分析酶促扩增的β-珠蛋白和HLA-DQα DNA。
Nature. 1986;324(6093):163-6. doi: 10.1038/324163a0.
3
Sensitive non-radioactive dot-blot hybridization using DNA probes labelled with chelate group substituted psoralen and quantitative detection by europium ion fluorescence.使用螯合基团取代补骨脂素标记的DNA探针进行灵敏的非放射性斑点杂交以及通过铕离子荧光进行定量检测。
Nucleic Acids Res. 1988 Feb 11;16(3):1181-96. doi: 10.1093/nar/16.3.1181.
4
Sensitive detection of genes by sandwich hybridization and time-resolved fluorometry.通过夹心杂交和时间分辨荧光法对基因进行灵敏检测。
Mol Cell Probes. 1987 Jun;1(2):159-68. doi: 10.1016/0890-8508(87)90024-7.
5
Time-resolved fluorometry for the identification of viral DNA in clinical specimens.用于鉴定临床标本中病毒DNA的时间分辨荧光测定法。
J Clin Microbiol. 1988 Nov;26(11):2434-6. doi: 10.1128/jcm.26.11.2434-2436.1988.
6
Deletion screening of the Duchenne muscular dystrophy locus via multiplex DNA amplification.通过多重DNA扩增对杜氏肌营养不良基因座进行缺失筛查。
Nucleic Acids Res. 1988 Dec 9;16(23):11141-56. doi: 10.1093/nar/16.23.11141.
7
Amplification of human minisatellites by the polymerase chain reaction: towards DNA fingerprinting of single cells.通过聚合酶链反应扩增人类小卫星：迈向单细胞DNA指纹识别
Nucleic Acids Res. 1988 Dec 9;16(23):10953-71. doi: 10.1093/nar/16.23.10953.
8
Generation of single-stranded DNA by the polymerase chain reaction and its application to direct sequencing of the HLA-DQA locus.通过聚合酶链反应生成单链DNA及其在HLA - DQA基因座直接测序中的应用。
Proc Natl Acad Sci U S A. 1988 Oct;85(20):7652-6. doi: 10.1073/pnas.85.20.7652.
9
Enzymatic amplification of beta-globin genomic sequences and restriction site analysis for diagnosis of sickle cell anemia.用于镰状细胞贫血诊断的β-珠蛋白基因组序列的酶促扩增及限制性酶切位点分析。
Science. 1985 Dec 20;230(4732):1350-4. doi: 10.1126/science.2999980.
10
Preparation of crude cell extract suitable for amplification of RNA by the polymerase chain reaction.制备适用于通过聚合酶链反应扩增RNA的粗细胞提取物。
Nucleic Acids Res. 1989 Mar 11;17(5):2141. doi: 10.1093/nar/17.5.2141.
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