Research Topics
| C T WittwerSummaryAffiliation: University of Utah Country: USA Publications
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Detail Information
Publications
High-resolution genotyping by amplicon melting analysis using LCGreenCarl T Wittwer
Department of Pathology, University of Utah Medical School, Salt Lake City, UT 84132, USA
Clin Chem 49:853-60. 2003..Our aim was to develop a closed-tube system for genotyping and mutation scanning that did not require labeled oligonucleotides...- High-resolution DNA melting analysis: advancements and limitationsCarl T Wittwer
Department of Pathology, University of Utah School of Medicine, Salt Lake City, UT 84132, USA
Hum Mutat 30:857-9. 2009.... - Real-time multiplex PCR assaysC T Wittwer
Department of Pathology, University of Utah School of Medicine, Salt Lake City, Utah 84132, USA
Methods 25:430-42. 2001..Instead of physical separation along the X and Y axes, amplification products are identified by different fluorescence spectra and melting characteristics... - High resolution melting analysis for gene scanningMaria Erali
ARUP Institute for Clinical and Experimental Pathology, Salt Lake City, UT 84108, USA
Methods 50:250-61. 2010..For each step in the protocol, a general overview of principles is provided, followed by an in depth analysis of one example, scanning of CYBB, the gene that is mutated in X-linked chronic granulomatous disease... - Amplicon melting analysis with labeled primers: a closed-tube method for differentiating homozygotes and heterozygotesCameron N Gundry
Department of Pathology, University of Utah Medical School, Salt Lake City, UT 84132, USA
Clin Chem 49:396-406. 2003..CONCLUSION: High-resolution melting analysis of PCR products amplified with labeled primers can identify both heterozygous and homozygous sequence variants... - Unlabeled oligonucleotide probes modified with locked nucleic acids for improved mismatch discrimination in genotyping by melting analysisLan-Szu Chou
ARUP Laboratories, Salt Lake City, UT 84108-1221, USA
Biotechniques 39:644, 646, 648 passim. 2005 - High-resolution DNA melting analysis for simultaneous mutation scanning and genotyping in solutionLuming Zhou
Department of Pathology, University of Utah Medical Center, Salt Lake City, UT 84105, USA
Clin Chem 51:1770-7. 2005..The method is no more complex than PCR and may reduce the need for resequencing... - Continuous fluorescence monitoring of rapid cycle DNA amplificationC T Wittwer
Department of Pathology, University of Utah Medical School, Salt Lake City 84132, USA
Biotechniques 22:130-1, 134-8. 1997..Continuous within-cycle monitoring allows rapid optimization of amplification conditions and should be particularly useful in developing new, standardized clinical assays... - Scanning the cystic fibrosis transmembrane conductance regulator gene using high-resolution DNA melting analysisJesse Montgomery
Department of Pathology, University of Utah Health Sciences Center, Salt Lake City, UT 84132, USA
Clin Chem 53:1891-8. 2007..High-resolution DNA melting analysis is a rapid, closed-tube alternative for gene scanning and genotyping... - Characterization of aberrant melting peaks in unlabeled probe assaysShale Dames
ARUP Institute for Clinical and Experimental Pathology, Salt Lake City, UT 84108, USA
J Mol Diagn 9:290-6. 2007.... - Amplicon DNA melting analysis for mutation scanning and genotyping: cross-platform comparison of instruments and dyesMark G Herrmann
Institute for Clinical and Experimental Pathology, ARUP, Salt Lake City, UT, USA
Clin Chem 52:494-503. 2006..The comparative performance of other instruments and dyes has not been evaluated... - Base-pair neutral homozygotes can be discriminated by calibrated high-resolution melting of small ampliconsCameron N Gundry
Idaho Technology Inc, 390 Wakara Way and Department of Pathology, University of Utah School of Medicine, 50 North Medical Drive 5B426, Salt Lake City, Utah 84108, USA
Nucleic Acids Res 36:3401-8. 2008..Unexpectedly, we were able to distinguish these homozygotes by T(m) even though current nearest neighbor models predict that the two homozygous alleles would be identical... - The LightCycler: a microvolume multisample fluorimeter with rapid temperature controlC T Wittwer
Idaho Technology, Idaho Falls, USA
Biotechniques 22:176-81. 1997..Complete amplification and analysis requires only 10-15 min... - RET proto-oncogene genotyping using unlabeled probes, the masking technique, and amplicon high-resolution melting analysisRebecca L Margraf
Advanced Technology Group, ARUP Institute for Clinical and Experimental Pathology, 500 Chipeta Way, Salt Lake City, UT 84108, USA
J Mol Diagn 9:184-96. 2007..Amplicon high-resolution melting analysis with unlabeled probes and the masking technique is a fast, accurate method for genotyping the >50 RET sequence variations... - Closed-tube genotyping of apolipoprotein E by isolated-probe PCR with multiple unlabeled probes and high-resolution DNA melting analysisMatthew Dean Poulson
Department of Pathology, University of Utah, Salt Lake City 84132, USA
Biotechniques 43:87-91. 2007..The IP-PCR technique should also be useful in detecting SNPs in other high-GC regions of the human genome... - Solution-phase DNA mutation scanning and SNP genotyping by nanoliter melting analysisScott O Sundberg
Department of Bioengineering, University of Utah, SLC, UT 84112, USA
Biomed Microdevices 9:159-66. 2007..Microscale systems for performing DNA melting reduce the reagents/DNA template required with a promise for high throughput analysis in a closed chamber without risk of contamination... - Expanded instrument comparison of amplicon DNA melting analysis for mutation scanning and genotypingMark G Herrmann
Institute for Clinical and Experimental Pathology, ARUP, Salt Lake City, UT 84108, USA
Clin Chem 53:1544-8. 2007..We assessed the performance of these new instruments for genotyping and scanning for mutations... - Unlabeled oligonucleotides as internal temperature controls for genotyping by amplicon meltingMichael T Seipp
ARUP Institute for Clinical and Experimental, Pathology, Salt Lake City, UT 84108, USA
J Mol Diagn 9:284-9. 2007..In conclusion, internal temperature controls increase the accuracy of genotyping by high-resolution amplicon melting and should also improve results on lower resolution instruments... - Enrichment and detection of rare alleles by means of snapback primers and rapid-cycle PCRLuming Zhou
Department of Pathology, University of Utah Health Sciences Center, Salt Lake City, UT 84132, USA
Clin Chem 56:814-22. 2010..Selective amplification of minority alleles is often necessary to detect cancer mutations in clinical samples... - LightCycler technology in molecular diagnosticsElaine Lyon
Department of Pathology, University of Utah, Salt Lake City, Utah, USA
J Mol Diagn 11:93-101. 2009..High-resolution melting allows mutation scanning by detecting all heterozygous changes. This review describes the major advances throughout the last 15 years regarding LightCycler technology and its application in clinical laboratories... - Rapid genetic analysis of x-linked chronic granulomatous disease by high-resolution meltingHarry R Hill
Department of Pathology, Pediatrics and Medicine, 5B114, University of Utah School of Medicine, 50 N Medical Dr, Salt Lake City, UT 84132, USA
J Mol Diagn 12:368-76. 2010..High-resolution melting can provide timely diagnosis at low cost for effective clinical management of rare, genetic primary immunodeficiency disorders... - Family clusters of variant X-linked chronic granulomatous diseaseJeffrey M Bender
Department of Pediatrics, University of Utah School of Medicine, Salt Lake City, Utah, USA
Pediatr Infect Dis J 28:529-33. 2009..A high index of suspicion in the setting of unusual infections such as Burkholderia cepacia pneumonia is essential to make the diagnosis. Family screening can lead to early intervention, prophylaxis, and appropriate genetic counseling... - Spinning disk platform for microfluidic digital polymerase chain reactionScott O Sundberg
University of Utah, Rm 5R441, 1795 E South Campus Dr, Salt Lake City, Utah 84112, USA
Anal Chem 82:1546-50. 2010..The spinning disk platform reduces disposable cost, instrument complexity, and thermocycling time compared to other current digital PCR platforms... - High resolution melting applications for clinical laboratory medicineMaria Erali
ARUP Institute for Clinical and Experimental Pathology, University of Utah, Salt Lake City, UT 84108, USA
Exp Mol Pathol 85:50-8. 2008..In the clinical laboratory, it is an ideal format for in-house testing, with minimal cost and time requirements for new assay development... - Quantitative heteroduplex analysis for single nucleotide polymorphism genotypingRobert A Palais
Department of Mathematics, University of Utah, Salt Lake City, UT 84112, USA
Anal Biochem 346:167-75. 2005..Optimal mixing before PCR followed by high-resolution melting analysis permits genotyping of all SNPs with a single closed-tube analysis... - Real-time PCR technology for cancer diagnosticsPhilip S Bernard
Department of Pathology, University of Utah School of Medicine, 30 North 1900 East, Salt Lake City 84132, USA
Clin Chem 48:1178-85. 2002..The challenge for pathology will be the development and implementation of these molecular classifications for routine clinical practice... - Distinguishing different DNA heterozygotes by high-resolution meltingRobert Graham
Department of Pathology, University of Utah School of Medicine, Salt Lake City 84132, USA
Clin Chem 51:1295-8. 2005 - Quantification of low-copy transcripts by continuous SYBR Green I monitoring during amplificationT B Morrison
Department of Pathology, University of Utah Medical School, Salt Lake City 84132, USA
Biotechniques 24:954-8, 960, 962. 1998..Above 10 copies per reaction, typical replicate coefficients of variation were 6%-37%, with better precision at higher copy numbers... - Quantification of HER2/neu gene amplification by competitive pcr using fluorescent melting curve analysisE Lyon
Department of Pathology, School of Medicine, University of Utah, Salt Lake 84132, USA
Clin Chem 47:844-51. 2001..We designed a quantitative PCR system utilizing fluorescent hybridization probes and a competitor that differed from the HER2/neu sequence by a single base change... - A comparison of high-resolution melting analysis with denaturing high-performance liquid chromatography for mutation scanning: cystic fibrosis transmembrane conductance regulator gene as a modelLan-Szu Chou
Institute for Clinical and Experimental Pathology, ARUP Laboratories, Salt Lake City, UT 84108-1221, USA
Am J Clin Pathol 124:330-8. 2005..HRMA had better sensitivity and specificity than dHPLC with the added advantage that some homozygous sequence alterations could be identified. HRMA has great potential for rapid, closed-tube mutation scanning... - Detection of c-kit-activating mutations in gastrointestinal stromal tumors by high-resolution amplicon melting analysisCarlynn Willmore
Department of Pathology, University of Utah, Salt Lake City 84108, USA
Am J Clin Pathol 122:206-16. 2004..High-resolution melting analysis can be used to scan DNA for potential c-kit-activating mutations and can aid in the diagnosis of GISTs... - Unlabeled probes for the detection and typing of herpes simplex virusShale Dames
ARUP Institute for Clinical and Experimental Pathology, Salt Lake City, UT 84108, USA
Clin Chem 53:1847-54. 2007..Unlabeled probes and amplicon melting have been used to detect small deletions and single-nucleotide polymorphisms in assays where template is in abundance. Unlabeled probes have not been applied to low-level target detection, however... - Rapid diagnosis of MEN2B using unlabeled probe melting analysis and the LightCycler 480 instrumentRebecca L Margraf
ARUP Institute for Clinical and Experimental Pathology, University of Utah Medical School, Salt Lake City, Utah, USA
J Mol Diagn 10:123-8. 2008..This assay demonstrates 100% specificity and sensitivity for the identification of RET mutations causative of MEN2B... - Sensitivity and specificity of single-nucleotide polymorphism scanning by high-resolution melting analysisGudrun H Reed
Department of Pathology, University of Utah Medical Center, 5B418, 50 N Medical Dr, Salt Lake City 84132, USA
Clin Chem 50:1748-54. 2004..The error rate of scanning depends on the PCR product size and the type of base change, but not on the position of the SNP. The technique requires only PCR reagents, the dye LCGreen I, and 1-2 min of closed-tube, post-PCR analysis... - The use of EGFR exon 19 and 21 unlabeled DNA probes to screen for activating mutations in non-small cell lung cancerCarlynn Willmore-Payne
Associated Regional and University Pathologists Institute for Clinical and Experimental Patholog y, Salt Lake City, UT 84108, USA
J Biomol Tech 19:217-24. 2008..This allows for easy identification of activating mutations even in homozygous or amplified states and is useful in the screening of NSCLC for the common EGFR activating mutations... - Closed-tube genotyping with unlabeled oligonucleotide probes and a saturating DNA dyeLuming Zhou
Department of Pathology, University of Utah School of Medicine, Salt Lake City, UT 84132, USA
Clin Chem 50:1328-35. 2004..The method is closed-tube, does not require fluorescently labeled probes or real-time PCR, and can be completed in <10 min on any instrument capable of monitoring melting curves by fluorescence... - Genotyping hepatitis C virus by heteroduplex mobility analysis using temperature gradient capillary electrophoresisRebecca L Margraf
ARUP Institute for Clinical and Experimental Pathology, Salt Lake City, Utah 84108, USA
J Clin Microbiol 42:4545-51. 2004..Compared to conventional HMA, TGCE improves the resolution, with better separation of heteroduplexes and homoduplexes. All common HCV genotypes can be detected and differentiated by this HMA-TGCE assay... - Real-time polymerase chain reaction and melting curve analysisRobert J Pryor
Department of Pathology, University of Utah School of Medicine, Salt Lake City, UT, USA
Methods Mol Biol 336:19-32. 2006..When used together with rapid temperature control, these methods allow amplification and genotyping in less than a half hour... - Evaluation of Her-2/neu gene status in osteosarcoma by fluorescence in situ hybridization and multiplex and monoplex polymerase chain reactionsCarlynn Willmore-Payne
Department of Pathology, University of Utah Health Sciences Center, Salt Lake City, UT 84132, USA
Arch Pathol Lab Med 130:691-8. 2006.... - Simultaneous mutation scanning and genotyping by high-resolution DNA melting analysisJesse Montgomery
Department of Pathology, UUMC, 5B418, 50 N Medical Drive, Salt Lake City, Utah 84105, USA
Nat Protoc 2:59-66. 2007..The PCR requires <30 min (capillaries) or 1.5 h (96- or 384-well plates) and melting acquisition takes 1-2 min per capillary or 5 min per plate... - A rapid and efficient method of genotyping zebrafish mutantsJohn M Parant
Department of Neurobiology and Anatomy, University of Utah School of Medicine, Salt Lake City, Utah 84112, USA
Dev Dyn 238:3168-74. 2009..This technique can genotype individual zebrafish embryos and adults (by tail-clip) and is applicable to other model organisms... - Plastic versus glass capillaries for rapid-cycle PCROluwole Elenitoba-Johnson
University of Utah, Salt Lake City, Utah 84132, USA
Biotechniques 44:487-8, 490, 492. 2008..Only the APOE target was successfully amplified by extending the denaturation and annealing times to 5 or 10 s. A 20 s holding period was necessary to reach target temperatures in plastic capillaries... - Masking selected sequence variation by incorporating mismatches into melting analysis probesRebecca L Margraf
ARUP Institute for Clinical and Experimental Pathology, Salt Lake City, Utah 84108, USA
Hum Mutat 27:269-78. 2006..The masking probes can also localize mutations to specific codons or nucleotide positions. Masking probes can simplify melting analysis of complex regions and eliminate the need for sequencing... - Rapid molecular analysis of the STAT3 gene in Job syndrome of hyper-IgE and recurrent infectious diseasesAttila Kumanovics
Department of Pathology, Human Genetics, University of Utah School of Medicine and the Associated Regional and University Pathologists Institute for Clinical and Experimental Pathology, Salt Lake City, UT, USA
J Mol Diagn 12:213-9. 2010.... - Human papillomavirus genotyping using an automated film-based chip arrayMaria Erali
ARUP Laboratories, 500 Chipeta Way, Salt Lake City, UT 84108, USA
J Mol Diagn 11:439-45. 2009..Based on its analytical performance, future studies with this platform are warranted to assess its clinical utility for HPV detection and genotyping... - Identifying common genetic variants by high-resolution meltingJoshua G Vandersteen
Department of Pathology, University of Utah Health Sciences Center, Salt Lake City, UT 84132, USA
Clin Chem 53:1191-8. 2007..Heteroduplex scanning techniques usually detect all heterozygotes, including common variants not of clinical interest... - Homogeneous amplification and mutation scanning of the p53 gene using fluorescent melting curvesHaleigh Millward
Idaho Technology Inc, Salt Lake City, UT 84105, USA
Clin Chem 48:1321-8. 2002..CONCLUSIONS: p53 mutation scanning by single-labeled hybridization probes is a homogeneous, rapid, and sensitive method with application in both research and clinical diagnostics... - Snapback primer genotyping with saturating DNA dye and melting analysisLuming Zhou
Department of Pathology, University of Utah Health Sciences Center, Salt Lake City, Utah 84132, USA
Clin Chem 54:1648-56. 2008..DNA hairpins have been used in molecular analysis of PCR products as self-probing amplicons. Either physical separation or fluorescent oligonucleotides with covalent modifications were previously necessary... - High-resolution DNA melting curve analysis to establish HLA genotypic identityL Zhou
Department of Pathology, University of Utah, Salt Lake City, 84132, USA
Tissue Antigens 64:156-64. 2004..The technique may also prove useful for confirmation of HLA genotypic identity between unrelated individuals prior to allogeneic hematopoietic stem-cell transplantation... - High-resolution DNA melting analysis for simple and efficient molecular diagnosticsGudrun H Reed
Department of Pathology, University of Utah Medical Center, 5B418, 50 North Medical Drive, Salt Lake City, UT 84132, USA
Pharmacogenomics 8:597-608. 2007..Owing to its simplicity and speed, the method is a good fit for personalized medicine as a rapid, inexpensive method to predict therapeutic response... - High-resolution DNA melting analysis in clinical research and diagnosticsJesse L Montgomery
Department of Pathology, University of Utah Health Sciences Center, Salt Lake City, UT 84132, USA
Expert Rev Mol Diagn 10:219-40. 2010.... - Mutation scanning of the RET protooncogene using high-resolution melting analysisRebecca L Margraf
Advanced Technology Group, ARUP Institute for Clinical and Experimental Pathology, Salt Lake City, UT 84108, USA
Clin Chem 52:138-41. 2006..The current widely used approach for RET mutation detection is sequencing of the exons... - Comparison of two quantitative polymerase chain reaction methods for detecting HER2/neu amplificationAlison Millson
Associated Regional and University Pathologists Institute for Clinical and Experimental Pathology, Salt Lake City, Utah, USA
J Mol Diagn 5:184-90. 2003..Real-time quantification and microdissection is useful clinically for HER2/neu quantification. Its ease of use and broad dynamic range allows screening for amplification of HER2/neu... - Mathematical algorithms for high-resolution DNA melting analysisRobert Palais
Department of Mathematics, University of Utah, Salt Lake City, Utah, USA
Methods Enzymol 454:323-43. 2009.... - Multiplex amplicon genotyping by high-resolution meltingMichael T Seipp
ARUP Institute for Clinical and Experimental Pathology, Salt Lake City, Utah 84108, USA
J Biomol Tech 20:160-4. 2009..The LS-32 simplifies more complex high-resolution melting assays by reducing hands-on manipulation, total time of analysis, and reagent cost while maintaining the resolution necessary for multiplex amplicon genotyping... - Molecular classification of melanoma using real-time quantitative reverse transcriptase-polymerase chain reactionTracey B Lewis
Research and Development, ARUP Laboratories Inc, Salt Lake City, Utah, USA
Cancer 104:1678-86. 2005..These markers could have diagnostic utility for the detection of melanoma micrometastasis in sentinel lymph nodes... - Quadruplex genotyping of F5, F2, and MTHFR variants in a single closed tube by high-resolution amplicon meltingMichael T Seipp
ARUP Institute for Clinical and Experimental Pathology, ARUP Laboratories, 500 Chipeta Way, Salt Lake City, UT 84108, USA
Clin Chem 54:108-15. 2008.... - Single-tube method for nucleic acid extraction, amplification, purification, and sequencingRebecca L Margraf
ARUP Institute for Clinical and Experimental Pathology, Salt Lake City, UT 84108, USA
Clin Chem 50:1755-61. 2004..We used a commercially available nucleic acid extraction system to develop a single-tube extraction-to-sequencing (STETS) method for HCV genotyping... - Rapid species identification within the Mycobacterium chelonae-abscessus group by high-resolution melting analysis of hsp65 PCR productsIan D Odell
Department of Pathology, University of Utah Medical School, Salt Lake City, 84132, USA
Am J Clin Pathol 123:96-101. 2005..Advantages of the method include speed, low risk of amplicon contamination (closed-tube), and no need for separation steps (sequencing, electrophoresis, high-performance liquid chromatography) or real-time monitoring... - Multiple endocrine neoplasia type 2 RET protooncogene database: repository of MEN2-associated RET sequence variation and reference for genotype/phenotype correlationsRebecca L Margraf
ARUP Institute for Clinical and Experimental Pathology R, Salt Lake City, Utah 84108, USA
Hum Mutat 30:548-56. 2009..The MEN2 RET database (www.arup.utah.edu/database/MEN2/MEN2_welcome.php) will serve as a repository for MEN2-associated RET sequence variation and reference for RET genotype/MEN2 phenotype correlations... - Instrument comparison for heterozygote scanning of single and double heterozygotes: a correction and extension of Herrmann et al., Clin Chem 2006;52:494-503Mark G Herrmann
Clin Chem 53:150-2. 2007 - Multiplex single-color PCR with amplicon melting analysis for identification of Aspergillus speciesMaria Erali
Clin Chem 52:1443-5. 2006 - Fifty years of molecular (DNA/RNA) diagnosticsThomas R Gingeras
Affymetrix, Inc, Santa Clara, CA, USA
Clin Chem 51:661-71. 2005
Research Grants
- HOMOGENEOUS MULTIPLEX PCR BY FLUORESCENCE AND TMCarl Wittwer; Fiscal Year: 2002..This will be the first real-time instrument compatible with all visible/near IR fluorescent probes and amplification techniques. Multiplexing by fluorescence and Tm greatly expands the power of real-time nucleic acid analysis. ..
- Homogeneous Mutation ScanningCarl Wittwer; Fiscal Year: 2006....
- A System for Rapid PCR, Mutation Scanning and GenotypingCarl Wittwer; Fiscal Year: 2006....