A HIGHLY SENSITIVE AND FAST NONRADIOACTIVE METHOD FOR DETECTION OF POLYMERASE CHAIN-REACTION PRODUCTS

被引：30

作者：

HOLMSTROM, K

ROSSEN, L

RASMUSSEN, OF

机构：

[1] Department of Molecular Food Technology, Biotechnological Institute, DK-2800 Lyngby, Lundtoftevej 100

来源：

ANALYTICAL BIOCHEMISTRY | 1993年 / 209卷 / 02期

关键词：

D O I：

10.1006/abio.1993.1120

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

A novel system for the detection of polymerase chain reaction (PCR) products has been developed. The system is based on a PCR in which one of the primers is biotinylated and digoxigenin-11-dUTP is incorporated during elongation. Biotinylated PCR products are captured on streptavidin-coated solid supports, and alkaline phosphatase conjugated to anti-digoxigenin antibody is subsequently bound to the incorporated digoxigenin. The detection may be obtained with colorimetric, fluorescent, or luminescent substrates for alkaline phosphatase. The detection system can be performed in microtiter plates allowing easy handling of multiple samples. The total assay time following the PCR is between 1 and 2 h dependent on the type of substrate and the type of solid support applied in the system. Within this period of time the system is capable of detecting 1 template in 29 cycles of PCR. © 1993 Academic Press, Inc.

引用

收藏

页码：278 / 283

页数：6

相关论文

共 22 条

[1] EFFICIENT METHODS FOR ATTACHING NONRADIOACTIVE LABELS TO THE 5' ENDS OF SYNTHETIC OLIGODEOXYRIBONUCLEOTIDES [J].

AGRAWAL, S ;

CHRISTODOULOU, C ;

GAIT, MJ .

NUCLEIC ACIDS RESEARCH, 1986, 14 (15) :6227-6245

[2] QUANTIFICATION OF DNA-SEQUENCES OBTAINED BY POLYMERASE CHAIN-REACTION USING A BIOLUMINESCENT ADSORBENT [J].

BALAGUER, P ;

TEROUANNE, B ;

BOUSSIOUX, AM ;

NICOLAS, JC .

ANALYTICAL BIOCHEMISTRY, 1991, 195 (01) :105-110

[3] DETECTION OF LISTERIA SPECIES AND LISTERIA-MONOCYTOGENES USING POLYMERASE CHAIN-REACTION [J].

BORDER, PM ;

HOWARD, JJ ;

PLASTOW, GS ;

SIGGENS, KW .

LETTERS IN APPLIED MICROBIOLOGY, 1990, 11 (03) :158-162

[4] MAXIMIZING SENSITIVITY AND SPECIFICITY OF PCR BY PREAMPLIFICATION HEATING [J].

DAQUILA, RT ;

BECHTEL, LJ ;

VIDELER, JA ;

ERON, JJ ;

GORCZYCA, P ;

KAPLAN, JC .

NUCLEIC ACIDS RESEARCH, 1991, 19 (13) :3749-3749

[5] DETECTION AND IDENTIFICATION OF LISTERIA-MONOCYTOGENES IN COOKED SAUSAGE PRODUCTS AND IN MILK BY INVITRO AMPLIFICATION OF HEMOLYSIN GENE FRAGMENTS [J].

FURRER, B ;

CANDRIAN, U ;

HOEFELEIN, C ;

LUETHY, J .

JOURNAL OF APPLIED BACTERIOLOGY, 1991, 70 (05) :372-379

[6] USE OF LABELED PRIMERS IN POLYMERASE CHAIN-REACTION (LP-PCR) FOR A RAPID DETECTION OF THE PRODUCT [J].

HAYASHI, K ;

ORITA, M ;

SUZUKI, Y ;

SEKIYA, T .

NUCLEIC ACIDS RESEARCH, 1989, 17 (09) :3605-3605

[7] SIMPLIFIED METHOD FOR CONFIRMATION OF PCR PRODUCTS [J].

KAI, M ;

KAMIYA, S ;

SAWAMURA, S ;

YAMAMOTO, T ;

OZAWA, A .

NUCLEIC ACIDS RESEARCH, 1991, 19 (16) :4562-4562

[8] DETECTION OF HEPATITIS-B VIRUS-DNA IN SERUM BY POLYMERASE CHAIN-REACTION AMPLIFICATION AND MICROTITER SANDWICH HYBRIDIZATION [J].

KELLER, GH ;

HUANG, DP ;

SHIH, JWK ;

MANAK, MM .

JOURNAL OF CLINICAL MICROBIOLOGY, 1990, 28 (06) :1411-1416

[9] QUANTITATIVE-ANALYSIS OF POLYMERASE CHAIN-REACTION (PCR) PRODUCTS USING PRIMERS LABELED WITH BIOTIN AND A FLUORESCENT DYE [J].

LANDGRAF, A ;

RECKMANN, B ;

PINGOUD, A .

ANALYTICAL BIOCHEMISTRY, 1991, 193 (02) :231-235

[10] CHEMILUMINESCENT NUCLEIC-ACID DETECTION WITH DIGOXIGENIN-LABELED PROBES - A MODEL SYSTEM WITH PROBES FOR ANGIOTENSIN CONVERTING ENZYME WHICH DETECT LESS THAN ONE ATTOMOLE OF TARGET DNA [J].

LANZILLO, JJ .

ANALYTICAL BIOCHEMISTRY, 1991, 194 (01) :45-53