| HOME | HELP | FEEDBACK | SUBSCRIPTIONS | ARCHIVE | SEARCH | TABLE OF CONTENTS |
From the Department of Pathology,
*
University of Miami, Miami, Florida; the Blood Center of Southeastern Wisconsin,
Milwaukee, Wisconsin; and the Department of Pathology and TriCore Reference Laboratories,
University of New Mexico, Albuquerque, New Mexico
The goal of this multicenter study was to evaluate the second-generation Invader technology for detecting the factor V (Leiden) mutation directly from genomic DNA of different sample types. Invader assay results were compared with polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) or allele-specific PCR (AS-PCR) analysis. The Invader assay is a PCR-independent methodology that uses a microtiter plate format. In the assay, a specific upstream Invader oligonucleotide and a downstream probe hybridize in tandem to a complementary DNA template and form a partially overlapping structure. The Cleavase VIII enzyme recognizes and cuts this structure to release the 5' flap of the probe. This flap then serves as an Invader oligonucleotide to direct cleavage of a fluorescence resonance energy transfer (FRET) probe in a second invasive cleavage reaction. Cleavage of this FRET probe results in the generation of a fluorescent signal. The results of the Invader assay were 99.5% concordant with the PCR-based methods. Of the 372 samples tested once, only two gave discordant results (one from operator error and one from unknown causes), but were concordant on retesting. These results indicate that a simple microtiter plate-based Invader assay can reliably genotype clinical patient samples for the factor V (Leiden) point mutation directly from genomic DNA without prior target amplification.
This article has been cited by other articles:
 |
|
 |
|
  J. J. Germer, D. W. Majewski, B. Yung, P. S. Mitchell, and J. D. C. Yao Evaluation of the Invader Assay for Genotyping Hepatitis C Virus J. Clin. Microbiol., February 1, 2006; 44(2): 318 - 323. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 W. Yang, T. Yaoi, S. Huang, Q. Yang, S. Hatcher, H. Seet, and J. P. Gregg Detecting the C282Y and H63D Mutations of the HFE Gene by Holliday Junction-Based Allele-Specific Genotyping Methods Clin. Chem., January 1, 2005; 51(1): 210 - 213. [Full Text] [PDF]
|
|
|
|
 |
|
 M. Patnaik, J. S. Dlott, R. N. Fontaine, M.T. Subbiah, M. J. Hessner, K. A. Joyner, M. R. Ledford, E. C. Lau, C. Moehlenkamp, J. Amos, et al. Detection of Genomic Polymorphisms Associated with Venous Thrombosis Using the Invader Biplex Assay J. Mol. Diagn., May 1, 2004; 6(2): 137 - 144. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 K. V. N. Rao, P. W. Stevens, J. G. Hall, V. Lyamichev, B. P. Neri, and D. M. Kelso Genotyping single nucleotide polymorphisms directly from genomic DNA by invasive cleavage reaction on microspheres Nucleic Acids Res., June 1, 2003; 31(11): e66 - e66. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
M. Erali, B. Schmidt, E. Lyon, and C. Wittwer Evaluation of Electronic Microarrays for Genotyping Factor V, Factor II, and MTHFR Clin. Chem., May 1, 2003; 49(5): 732 - 739. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
S. A. E. Marras, F. R. Kramer, and S. Tyagi Efficiencies of fluorescence resonance energy transfer and contact-mediated quenching in oligonucleotide probes Nucleic Acids Res., November 1, 2002; 30(21): e122 - e122. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
J. G. Evans and C. Lee-Tataseo Determination of the Factor V Leiden Single-Nucleotide Polymorphism in a Commercial Clinical Laboratory by Use of NanoChip Microelectronic Array Technology Clin. Chem., September 1, 2002; 48(9): 1406 - 1411. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
M. J. Hessner, K. D. Friedman, K. V. Voelkerding, S. Huber, D. Ryan, B. Nuccie, and M. Ledford Multisite Study for Genotyping of the Factor II (Prothrombin) G20210A Mutation by the Invader Assay Clin. Chem., November 1, 2001; 47(11): 2048 - 2050. [Full Text] [PDF]
|
|
| HOME | HELP | FEEDBACK | SUBSCRIPTIONS | ARCHIVE | SEARCH | TABLE OF CONTENTS |
