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.
journal_name
J Clin Microbioljournal_title
Journal of clinical microbiologyauthors
Dahlén PO,Iitiä AJ,Skagius G,Frostell A,Nunn MF,Kwiatkowski Mdoi
10.1128/JCM.29.4.798-804.1991subject
Has Abstractpub_date
1991-04-01 00:00:00pages
798-804issue
4eissn
0095-1137issn
1098-660Xjournal_volume
29pub_type
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journal_title:Journal of clinical microbiology
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journal_title:Journal of clinical microbiology
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journal_title:Journal of clinical microbiology
pub_type: 杂志文章
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journal_title:Journal of clinical microbiology
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doi:10.1128/JCM.35.9.2215-2219.1997
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journal_title:Journal of clinical microbiology
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pub_type: 杂志文章
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journal_title:Journal of clinical microbiology
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journal_title:Journal of clinical microbiology
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journal_title:Journal of clinical microbiology
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doi:10.1128/JCM.14.3.313-317.1981
更新日期:1981-09-01 00:00:00
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journal_title:Journal of clinical microbiology
pub_type: 杂志文章
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journal_title:Journal of clinical microbiology
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pub_type: 杂志文章
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journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/JCM.39.11.4058-4065.2001
更新日期:2001-11-01 00:00:00