LightCycler Technology in Molecular Diagnostics

被引:73
作者
Lyon, Elaine
Wittwer, Carl T.
机构
[1] Univ Utah, Dept Pathol, Salt Lake City, UT USA
[2] ARUP Inst Expt & Clin Pathol, Salt Lake City, UT USA
关键词
POLYMERASE-CHAIN-REACTION; DNA MELTING ANALYSIS; FACTOR-V-LEIDEN; REAL-TIME PCR; FLUORESCEIN-LABELED OLIGONUCLEOTIDES; GASTROINTESTINAL STROMAL TUMORS; KIT-ACTIVATING MUTATIONS; RAPID-CYCLE PCR; CURVE ANALYSIS; HEMOCHROMATOSIS MUTATIONS;
D O I
10.2353/jmoldx.2009.080094
中图分类号
R36 [病理学];
学科分类号
100104 ;
摘要
LightCycler technology combines rapid-cycle polymerase chain reaction with real-time fluorescent monitoring and melting curve analysis. Since its introduction in 1997, It is now used in many areas of molecular pathology, including oncology (solid tumors and hematopathology), inherited disease, and infectious disease. By monitoring product accumulation during rapid amplification, quantitative polymerase chain reaction in a closed-tube system is possible in 15 to 30 minutes. Furthermore, melting curve analysis of probes and/or amplicons provides genotyping and even haplotyping. Novel mutations are identified by unexpected melting temperature or curve shape changes. Melting probe designs include adjacent hybridization probes, single labeled probes, unlabeled probes, and snapback primers. High-resolution melting allows mutation scanning by detecting all heterozygous changes. This review describes the major advances throughout the last 1.5 years regarding LightCycler technology and its application in clinical laboratories.(j Moldx Diagn 2009, 11-9.3-101; DOI: 10.23531/jmoldx.2009.080094)
引用
收藏
页码:93 / 101
页数:9
相关论文
共 86 条
[1]  
*ACMG CAP MOL GEN, 2006, PART SUMM REP MGL B, P4
[2]   Randomized trial of genotype-guided versus standard warfarin dosing in patients initiating oral anticoagulation [J].
Anderson, Jeffrey L. ;
Horne, Benjamin D. ;
Stevens, Scott M. ;
Grove, Amanda S. ;
Barton, Stephanie ;
Nicholas, Zachery P. ;
Kahn, Samera F. S. ;
May, Heidi T. ;
Samuelson, Kent M. ;
Muhlestein, Joseph B. ;
Carlquist, John F. .
CIRCULATION, 2007, 116 (22) :2563-2570
[3]   Validation of high-resolution DNA melting analysis for mutation scanning of the cystic fibrosis transmembrane conductance regulator (CFTR) gene [J].
Audrezet, Marie-Pierre ;
Dabricot, Aurelia ;
Le Marechal, Cedric ;
Ferec, Claude .
JOURNAL OF MOLECULAR DIAGNOSTICS, 2008, 10 (05) :424-434
[4]   High-throughput amplicon scanning of the TP53 gene in breast cancer using high-resolution fluorescent melting curve analyses and automatic mutation calling [J].
Bastien, Roy ;
Lewis, Tracey B. ;
Hawkes, Jason E. ;
Quackenbush, John F. ;
Robbins, Thomas C. ;
Palazzo, Juan ;
Perou, Charles M. ;
Bernard, Philip S. .
HUMAN MUTATION, 2008, 29 (05) :757-764
[5]   Homogeneous multiplex genotyping of hemochromatosis mutations with fluorescent hybridization probes [J].
Bernard, PS ;
Ajioka, RS ;
Kushner, JP ;
Wittwer, CT .
AMERICAN JOURNAL OF PATHOLOGY, 1998, 153 (04) :1055-1061
[6]  
Bolufer P, 2000, HAEMATOLOGICA, V85, P1248
[7]  
BUSTIN S, CLIN CHEM IN PRESS
[8]   Unlabeled oligonucleotide probes modified with locked nucleic acids for improved mismatch discrimination in genotyping by melting analysis [J].
Chou, LS ;
Meadows, C ;
Wittwer, CT ;
Lyon, E .
BIOTECHNIQUES, 2005, 39 (05) :644-+
[9]  
Cockerill FR, 2003, ARCH PATHOL LAB MED, V127, P1112
[10]   Fluorescein-labeled oligonucleotides for real-time PCR: Using the inherent quenching of deoxyguanosine nucleotides [J].
Crockett, AO ;
Wittwer, CT .
ANALYTICAL BIOCHEMISTRY, 2001, 290 (01) :89-97