Detection of prolactin inducible protein mRNA, a biomarker for breast cancer metastasis, using a molecular beacon-based assay.
Mortality due to breast cancer is increasingly linked to early, undetected metastasis, making methods for earlier detection acutely necessary. We describe the development of an assay based on molecular beacon (MB) chemistry with fluorescence detection to monitor a breast cancer biomarker for the analysis of breast cancer metastasis. The MB assay is based on the complementary base-pairing interactions of the MB nucleic acid with mRNA indicative of breast cancer metastasis. The presence of mRNA is characterized by an increase in the fluorescence intensity of the molecular beacon. The assay gives a linear, reproducible response to prolactin inducible protein mRNA, with a limit of detection in the high picomolar range. This method sensitively and specifically identifies a biomarker directly in serum samples in minimal time and with a straightforward procedure, dramatically reducing the total time for sample analysis over current methods from days to hours. The potential impact of this work in detection and understanding of breast cancer metastasis lies in improvements in simplicity, accuracy, and speed over current methods, which could allow for improved patient treatment and prognoses. Ultimately, additional sample throughput will result in better understanding of disease progression.
Department of Chemistry, Kalamazoo College, 1200 Academy St., Kalamazoo, MI 49006, USA.
Erik D
ED
Guetschow
Will
W
Black
Carolyn M
CM
Walsh
Jennifer R W
JR
Furchak
D001483:Base Sequence; D014408:Biomarkers, Tumor / Q000032:analysis*; D001943:Breast Neoplasms / Q000175:diagnosis* / Q000235:genetics / Q000378:metabolism; D002352:Carrier Proteins / Q000235:genetics*; D017931:DNA Primers; D005260:Female; D006023:Glycoproteins / Q000235:genetics*; D006801:Humans; D026901:Membrane Transport Proteins; D016133:Polymerase Chain Reaction; D012333:RNA, Messenger / Q000032:analysis*
D014408:Biomarkers, Tumor; D002352:Carrier Proteins; D017931:DNA Primers; D006023:Glycoproteins; D026901:Membrane Transport Proteins; C488892:PIP protein, human; D012333:RNA, Messenger