NDLI: SNaPshot and StripAssay as valuable alternatives to direct sequencing for KRAS mutation detection in colon cancer routine diagnostics.

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Year: 2011 Volume: 13 Issue: 1

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SNaPshot and StripAssay as valuable alternatives to direct sequencing for KRAS mutation detection in colon cancer routine diagnostics.

Content Provider	World Health Organization (WHO)-Global Index Medicus
Author	Fariña Sarasqueta, Arantza Moerland, Elna de Bruyne, Hanneke de Graaf, Henk Vrancken, Tamara Van Lijnschoten, Gesina Van Den Brule, Adriaan J. C.
Description	Country affiliation: Netherlands Author Affiliation: Fariña Sarasqueta A ( PAMM Laboratory for Pathology, Eindhoven, the Netherlands.)
Abstract	Although direct sequencing is the gold standard for KRAS mutation detection in routine diagnostics, it remains laborious, time consuming, and not very sensitive. Our objective was to evaluate SNaPshot and the KRAS StripAssay as alternatives to sequencing for KRAS mutation detection in daily practice. KRAS exon 2-specific PCR followed by sequencing or by a SNaPshot reaction was performed. For the StripAssay, a mutant-enriched PCR was followed by hybridization to KRAS-specific probes bound to a nitrocellulose strip. To test sensitivities, dilution series of mutated DNA in wild-type DNA were made. Additionally, direct sequencing and SNaPshot were evaluated in 296 colon cancer samples. Detection limits of direct sequencing, SNaPshot, and StripAssay were 20%, 10%, and 1% tumor cells, respectively. Direct sequencing and SNaPshot can detect all 12 mutations in KRAS codons 12 and 13, whereas the StripAssay detects 10 of the most frequent ones. Workload and time to results are comparable for SNaPshot and direct sequencing. SNaPshot is flexible and easy to multiplex. The StripAssay is less time consuming for daily laboratory practice. SNaPshot is more flexible and slightly more sensitive than direct sequencing. The clinical evaluation showed comparable performances between direct sequencing and SNaPshot. The StripAssay is rapid and an extremely sensitive assay that could be considered when few tumor cells are available. However, found mutants should be confirmed to avoid risk of false positives.
File Format	HTM / HTML
ISSN	15251578
e-ISSN	19437811
DOI	10.1016/j.jmoldx.2010.10.006
Journal	The Journal of Molecular Diagnostics
Issue Number	2
Volume Number	13
Language	English
Publisher	Elsevier
Publisher Date	2011-03-01
Publisher Place	United States
Access Restriction	Open
Subject Keyword	Discipline Molecular Biology Colonic Neoplasms Dna, Neoplasm Mutation Polymerase Chain Reaction Proto-oncogene Proteins Genetics Ras Proteins Diagnosis Enzymology Proto-oncogene Proteins P21(ras) Reproducibility Of Results Sensitivity And Specificity Sequence Analysis, Dna Evaluation Studies
Content Type	Text
Resource Type	Article
Subject	Molecular Medicine Pathology and Forensic Medicine

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