Sensitivity of electrospray ionization mass spectrometry detection of codon 249 mutations in the p53 gene compared with RFLP

Geng Sun Qian, Shuang Yuan Kuang, Xia He, John Davis Groopman, Peta E. Jackson

Research output: Contribution to journalArticle

Abstract

Hepatocellular carcinoma (HCC) has several major etiological risk factors, including infection with hepatitis viruses and exposure to aflatoxin B1. A specific missense mutation resulting from a guanine to thymine transversion at the third position of codon 249 in the p53 tumor suppressor gene has been reported in 10-70% of HCCs from areas of high dietary exposure to aflatoxin B1. This mutation has not only been detected in tumor samples but has also been measured in DNA isolated from the blood of patients with HCC in two separate studies by two independent methods: RFLP and short oligonucleotide mass analysis (SOMA), an electrospray ionization mass spectrometry technique. To compare the relative sensitivities of these methodologies, a set of serially diluted samples was analyzed by both techniques. The detection limits of RFLP and SOMA were 6% and 2.4% mutant alleles in the presence of wild-type alleles, respectively. When the DNA samples were predigested with HaeIII before SOMA, the detection limit was improved to 0.4% mutant allele in the presence of wildtype alleles. We have therefore found that SOMA is about 2.5-15-fold more sensitive than RFLP for detection of specific p53 mutations. A set of 26 DNA samples from HCC and normal liver was analyzed by RFLP and SOMA, and 5 samples were positive for the p53 mutation. An additional 4 samples were found to be positive for the mutation when SOMA was repeated after HaeIII predigestion.

Original languageEnglish (US)
Pages (from-to)1126-1129
Number of pages4
JournalCancer Epidemiology Biomarkers and Prevention
Volume11
Issue number10 I
StatePublished - Oct 2002

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Electrospray Ionization Mass Spectrometry
p53 Genes
Codon
Oligonucleotides
Restriction Fragment Length Polymorphisms
Mutation
Alleles
Hepatocellular Carcinoma
Aflatoxin B1
Limit of Detection
DNA
Hepatitis Viruses
Thymine
Guanine
Missense Mutation
Tumor Suppressor Genes
Liver
Infection
Neoplasms

ASJC Scopus subject areas

  • Epidemiology
  • Oncology

Cite this

Sensitivity of electrospray ionization mass spectrometry detection of codon 249 mutations in the p53 gene compared with RFLP. / Qian, Geng Sun; Kuang, Shuang Yuan; He, Xia; Groopman, John Davis; Jackson, Peta E.

In: Cancer Epidemiology Biomarkers and Prevention, Vol. 11, No. 10 I, 10.2002, p. 1126-1129.

Research output: Contribution to journalArticle

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abstract = "Hepatocellular carcinoma (HCC) has several major etiological risk factors, including infection with hepatitis viruses and exposure to aflatoxin B1. A specific missense mutation resulting from a guanine to thymine transversion at the third position of codon 249 in the p53 tumor suppressor gene has been reported in 10-70{\%} of HCCs from areas of high dietary exposure to aflatoxin B1. This mutation has not only been detected in tumor samples but has also been measured in DNA isolated from the blood of patients with HCC in two separate studies by two independent methods: RFLP and short oligonucleotide mass analysis (SOMA), an electrospray ionization mass spectrometry technique. To compare the relative sensitivities of these methodologies, a set of serially diluted samples was analyzed by both techniques. The detection limits of RFLP and SOMA were 6{\%} and 2.4{\%} mutant alleles in the presence of wild-type alleles, respectively. When the DNA samples were predigested with HaeIII before SOMA, the detection limit was improved to 0.4{\%} mutant allele in the presence of wildtype alleles. We have therefore found that SOMA is about 2.5-15-fold more sensitive than RFLP for detection of specific p53 mutations. A set of 26 DNA samples from HCC and normal liver was analyzed by RFLP and SOMA, and 5 samples were positive for the p53 mutation. An additional 4 samples were found to be positive for the mutation when SOMA was repeated after HaeIII predigestion.",
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