Comparison of real-time PCR-optimized oligonucleotide probe method and Sanger sequencing for detection of KRAS mutations in colorectal and lung carcinomas / 中华病理学杂志
Chinese Journal of Pathology
; (12): 599-602, 2012.
Article
de Zh
| WPRIM
| ID: wpr-303511
Bibliothèque responsable:
WPRO
ABSTRACT
<p><b>OBJECTIVE</b>To investigate the feasibility of real-time PCR-optimized oligonucleotide probe method for detection of KRAS mutations in lung and colorectal carcinomas, as compared with Sanger sequencing method.</p><p><b>METHODS</b>Genomic DNA was extracted from formalin fixed paraffin embedded samples of 221 lung carcinomas and 131 colorectal carcinomas after tumor cell content assessment and macrodissection. Real-time PCR-optimized oligonucleotide probe method and Sanger sequencing were performed to detect KRAS gene mutations. The frequency of KRAS mutation, mutation types, and their concordance were analyzed.</p><p><b>RESULTS</b>KRAS mutation was detected in 6.3% (14/221) and 4.5% (10/221) of 221 lung cancer samples by using real-time PCR-optimized oligonucleotide probe method and Sanger sequencing, respectively, while in 41.2% (54/131) and 40.5% (53/131) of 131 colorectal cancer samples, respectively. There was no significant correlation between KRAS gene mutations and patients' gender and age (P > 0.05). The positive rate of KRAS codon 12 mutation was significantly higher than that of KRAS codon 13 mutation (P < 0.05). The overall concordance between real-time PCR-optimized oligonucleotide probe method and Sanger sequencing for KRAS mutation detection was 97.4%.</p><p><b>CONCLUSION</b>Real-time PCR-optimized oligonucleotide probe method provides an alternative method with high consistency and sensitivity as compared to Sanger sequencing in gene mutation detection.</p>
Texte intégral:
1
Indice:
WPRIM
Sujet Principal:
Codon
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Analyse de mutations d'ADN
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Tumeurs colorectales
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Sondes oligonucléotidiques
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Protéines proto-oncogènes p21(ras)
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Protéines proto-oncogènes
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Sensibilité et spécificité
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Protéines G ras
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Réaction de polymérisation en chaine en temps réel
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Génétique
Type d'étude:
Diagnostic_studies
Limites du sujet:
Female
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Humans
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Male
langue:
Zh
Texte intégral:
Chinese Journal of Pathology
Année:
2012
Type:
Article