title: Rapid detection of SNP (c.309T>G) in the MDM2 gene by the Duplex SmartAmp method.
abstract: BACKGROUND
  Genetic polymorphisms in the human MDM2 gene are suggested to be a tumor susceptibility marker and a prognostic factor for cancer. It has been reported that a single nucleotide polymorphism (SNP) c.309T>G in the MDM2 gene attenuates the tumor suppressor activity of p53 and accelerates tumor formation in humans.
  
  
  METHODS
  In this study, to detect the SNP c.309T>G in the MDM2 gene, we have developed a new SNP detection method, named "Duplex SmartAmp," which enabled us to simultaneously detect both 309T and 309G alleles in one tube. To develop this new method, we introduced new primers i.e., nBP and oBPs, as well as two different fluorescent dyes that separately detect those genetic polymorphisms.
  
  
  CONCLUSIONS
  By the Duplex SmartAmp method, the genetic polymorphisms of the MDM2 gene were detected directly from a small amount of genomic DNA or blood samples. We used 96 genomic DNA and 24 blood samples to validate the Duplex SmartAmp by comparison with results of the conventional PCR-RFLP method; consequently, the Duplex SmartAmp results agreed totally with those of the PCR-RFLP method. Thus, the new SNP detection method is considered useful for detecting the SNP c.309T>G in the MDM2 gene so as to judge cancer susceptibility against some cellular stress in the clinical setting, and also to handle a large number of samples and enable rapid clinical diagnosis.
authors.affiliation: Division of Thoracic and Visceral Organ Surgery, Gunma University Graduate School of Medicine, Maebashi, Gunma, Japan.
authors.forename: Yasuaki
authors.initials: Y
authors.lastname: Enokida
authors.forename: Kimihiro
authors.initials: K
authors.lastname: Shimizu
authors.forename: Jun
authors.initials: J
authors.lastname: Atsumi
authors.forename: Alexander
authors.initials: A
authors.lastname: Lezhava
authors.forename: Yuki
authors.initials: Y
authors.lastname: Tanaka
authors.forename: Yasumasa
authors.initials: Y
authors.lastname: Kimura
authors.forename: Takahiro
authors.initials: T
authors.lastname: Soma
authors.forename: Takeshi
authors.initials: T
authors.lastname: Hanami
authors.forename: Yuki
authors.initials: Y
authors.lastname: Kawai
authors.forename: Kengo
authors.initials: K
authors.lastname: Usui
authors.forename: Yasuko
authors.initials: Y
authors.lastname: Okano
authors.forename: Seiichi
authors.initials: S
authors.lastname: Kakegawa
authors.forename: Hiroomi
authors.initials: H
authors.lastname: Ogawa
authors.forename: Yohei
authors.initials: Y
authors.lastname: Miyamae
authors.forename: Yohei
authors.initials: Y
authors.lastname: Miyagi
authors.forename: Haruhiko
authors.initials: H
authors.lastname: Nakayama
authors.forename: Toshihisa
authors.initials: T
authors.lastname: Ishikawa
authors.forename: Yoshihide
authors.initials: Y
authors.lastname: Hayashizaki
authors.forename: Izumi
authors.initials: I
authors.lastname: Takeyoshi
mesh_terms: D000368:Aged; D000483:Alleles; D005260:Female; D005787:Gene Frequency; D020022:Genetic Predisposition to Disease; D005838:Genotype; D006801:Humans; D008297:Male; D008875:Middle Aged; D009369:Neoplasms / Q000235:genetics / Q000473:pathology; D021141:Nucleic Acid Amplification Techniques / Q000379:methods; D016133:Polymerase Chain Reaction / Q000379:methods; D012150:Polymorphism, Restriction Fragment Length; D020641:Polymorphism, Single Nucleotide; D051736:Proto-Oncogene Proteins c-mdm2; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity
chemical_list: C497206:MDM2 protein, human; D051736:Proto-Oncogene Proteins c-mdm2
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