ArticleWorld journal of gastroenterology2010
Comparative analysis of dideoxy sequencing, the KRAS StripAssay and pyrosequencing for detection of KRAS mutation.
Article in World journal of gastroenterology, 2010. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 14 papers.
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Who cites it
14 citing papers in PubMed, 24 citations in OpenAlex.
- Review
- [Advances in Diagnosis and Targeted Therapy of KRASG12C Mutant Non-small Cell Lung Cancer].Zhongguo fei ai za zhi = Chinese journal of lung cancer · 2025Review
- Clinical retrospective study on the expression of the PD-L1 molecule in sporadic colorectal cancer and its correlation with K-ras gene mutations in Chinese patients.American journal of translational research · 2021Article
- Not all mutations ofInternational journal of clinical and experimental pathology · 2019Article
- Current companion diagnostics in advanced colorectal cancer; getting a bigger and better piece of the pie.Journal of gastrointestinal oncology · 2017Review
- Article
- Article
- Frequency of KRAS mutations in adult Korean patients with acute myeloid leukemia.International journal of hematology · 2013Article
- 454 next generation-sequencing outperforms allele-specific PCR, Sanger sequencing, and pyrosequencing for routine KRAS mutation analysis of formalin-fixed, paraffin-embedded samples.OncoTargets and therapy · 2013Article
- A comparison of Direct sequencing, Pyrosequencing, High resolution melting analysis, TheraScreen DxS, and the K-ras StripAssay for detecting KRAS mutations in non small cell lung carcinomas.Journal of experimental & clinical cancer research : CR · 2012Article
- KRAS Testing: A Tool for the Implementation of Personalized Medicine.Genes & cancer · 2012Article
- Analytical performance of a PCR assay for the detection of KRAS mutations (codons 12/13 and 61) in formalin-fixed paraffin-embedded tissue samples of colorectal carcinoma.Virchows Archiv : an international journal of pathology · 2012Article
- Clinical significance of K-ras and BRAF mutations in Chinese colorectal cancer patients.World journal of gastroenterology · 2011Article
- KRAS mutation detection in paired frozen and Formalin-Fixed Paraffin-Embedded (FFPE) colorectal cancer tissues.International journal of molecular sciences · 2011Article
Corrections and comments
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Authors and funding
4 authors at 1 institution in 1 country.
Funding
No grant is acknowledged in the PubMed record.
Abstract
aimTo compare the differences between dideoxy sequencing/KRAS StripAssay/pyrosequencing for detection of KRAS mutation in Chinese colorectal cancer (CRC) patients.
methodsFormalin-fixed, paraffin-embedded (FFPE) samples with tumor cells ≥ 50% were collected from 100 Chinese CRC patients at Beijing Cancer Hospital. After the extraction of genome DNA from FFPE samples, fragments contained codons 12 and 13 of KRAS exon 2 were amplified by polymerase chain reaction and analyzed by dideoxy sequencing, the KRAS StripAssay and pyrosequencing. In addition, the sensitivities of the 3 methods were compared on serial dilutions (contents of mutant DNA: 100%, 50%, 20%, 15%, 10%, 5%, 1%, 0%) of A549 cell line DNA (carrying the codon 12 Gly>Ser mutation) into wild-type DNA (human normal intestinal mucosa). The results of dideoxy sequencing, the KRAS StripAssay and pyrosequencing were analyzed by Chromas Software, Collector for KRAS StripAssay and the pyrosequencing PyroMarkTM Q24 system, respectively.
resultsAmong 100 patients, KRAS mutations were identified in 34%, 37% and 37% of patients by dideoxy sequencing, the KRAS StripAssay and pyrosequencing, respectively. The sensitivity was highest with the KRAS StripAssay (1%), followed by pyrosequencing (5%), and dideoxy sequencing was lowest (15%). Six different mutation types were found in this study with 3 main mutations Gly12Asp (GGT>GAT), Gly12Val (GGT>GTT) and Gly13Asp (GGC>GAC). Thirty-three patients were identified to have KRAS mutations by the 3 methods, and a total of 8 patients had conflicting results between 3 methods: 4 mutations not detected by dideoxy sequencing and the KRAS StripAssay were identified by pyrosequencing; 3 mutations not detected by dideoxy sequencing and pyrosequencing were identified by the KRAS StripAssay; and 1 mutation not detected by pyrosequencing was confirmed by dideoxy sequencing and the KRAS StripAssay. Among these discordant results, the results identified by dideoxy sequencing were consistent either with the KRAS StripAssay or with pyrosequencing, which indicated that the accuracy of dideoxy sequencing was high.
conclusionTaking a worldwide view of reports and our results, dideoxy sequencing remains the most popular method because of its low cost and high accuracy.
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