P53 assay for a urine test for HCC screening

摘要

A rapid and sensitive assay to detect p53 mutations in urine has been developed for use in screening cancer patients. The method uses a locked nucleic acid (LNA) clamp mediated one-step PCR-based assay with a sensitivity of up to a single copy and can be used not only in urine, but also other biological samples. The assay is particularly useful for hepatocellular carcinoma, colon cancer, breast cancer, lung cancer, prostate cancer, ovarian cancer, bladder cancer, lymphoma, and stomach cancer.

主权项

1. A method for detecting hepatocellular carcinoma in a subject comprising:
amplifying a low molecular weight p53 249T mutation in a urine sample from said subject, wherein the amplification comprises:
suppressing the amplification of wildtype p53 nucleic acid sequences using a Locked Nucleic Acid clamp formed of GGAGGCC, wherein the Locked Nucleic Acid clamp binds wildtype p53 but not p53 249T nucleic acid sequences, andamplifying the sample with primers consisting of SEQ ID NO: 1 and SEQ ID NO: 3; and detecting the presence of p53 249T mutant using an oligonucleotide probe, wherein the presence of p53 249T mutant nucleic acid sequences is indicative of hepatocellular carcinoma.