THE USE OF EUROPIUM (EU-3+) LABELED PRIMERS IN PCR AMPLIFICATION OF SPECIFIC TARGET DNA

被引：23

作者：

DAHLEN, P ^{[1
]}

IITIA, A ^{[1
]}

MUKKALA, VM ^{[1
]}

HURSKAINEN, P ^{[1
]}

KWIATKOWSKI, M ^{[1
]}

机构：

[1] WALLAC OY,SF-20101 TURKU,FINLAND

来源：

MOLECULAR AND CELLULAR PROBES | 1991年 / 5卷 / 02期

关键词：

TIME-RESOLVED FLUOROMETRY; EUROPIUM; POLYMERASE CHAIN REACTION (PCR); LABELED PRIMERS; NESTED PRIMERS; HIV-1;

D O I：

10.1016/0890-8508(91)90009-9

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

The polymerase chain reaction (PCR) has many potential applications in the field of DNA probe diagnostics. Here we describe a method that utilizes PCR and time-resolved fluorometry (TRF) for the detection of specific target DNA. First the DNA segment to be detected is amplified according to standard procedures. Then a pair of europium (Eu3+) and biotin-labelled primers nested within the amplified fragment is incorporated in a few additional PCR cycles. Thus amplified DNA fragments are generated that contain an affinity label (biotin) and a detectable label (europium). The doubly-labelled amplified DNA fragments are collected onto streptavidin coated microtitration strips and the bound Eu3+ is measured in a time-resolved fluorometer. We show here the application of this method to the detection of HIV-1 DNA. As few as five copies of HIV-1 DNA could readily be detected using this assay. The method described here is sensitive, rapid and easy to employ. In addition it lends itself to automation. © 1991.

引用

页码：143 / 149

页数：7

共 21 条

[1] DISEASE DIAGNOSIS BY RECOMBINANT-DNA METHODS [J].