Amplified telomerase analysis by using rotating magnetic particles: the rapid and sensitive detection of cancer cells.
A highly sensitive telomerase detection method that involves amplified telomerase analysis and the use of rotating magnetic particles has been developed. Magnetic particles, functionalized with a primer (1) that is recognized by telomerase, are mixed with a nucleotide mixture that includes biotinylated-dUTP, and telomerase-induced elongation of the primers proceeds with simultaneous biotin incorporation. Avidin-Horseradish peroxidase conjugate, coupled to biotin labels, yields the biocatalytic functional particles. Mixing the resulting particles with naphthoquinone-modified magnetic particles enables the optoelectronic detection of telomerase. Attraction of the magnetic particles to an electrode, followed by rotation of the particles, causes the electrocatalytic reduction of O(2) to H(2)O(2) and HRP-catalyzed oxidation of luminol (3); this results in chemilumunescence. The intensity of the emitted light depends on the telomerase content of the sample and the rotation speed of the particles. A minimum number of 10 cancer cells could be detected.
Weizmann
Yossi
Y
Institute of Chemistry, The Hebrew University of Jerusalem, Jerusalem 91904, Israel.
Patolsky
Fernando
F
Katz
Eugenii
E
Willner
Itamar
I
D000230:Adenocarcinoma / Q000175:diagnosis* / Q000378:metabolism; D015374:Biosensing Techniques / Q000295:instrumentation / Q000379:methods*; D019921:Biotinylation; D002294:Carcinoma, Squamous Cell / Q000175:diagnosis* / Q000378:metabolism; D002478:Cells, Cultured; D004247:DNA / Q000032:analysis*; D006367:HeLa Cells; D006735:Horseradish Peroxidase / Q000378:metabolism; D006801:Humans; D006861:Hydrogen Peroxide / Q000032:analysis; D007668:Kidney / Q000378:metabolism; D008163:Luminescent Measurements; D008165:Luminol / Q000737:chemistry; D008175:Lung Neoplasms / Q000175:diagnosis* / Q000378:metabolism; D008280:Magnetics; D009285:Naphthoquinones / Q000737:chemistry; D012680:Sensitivity and Specificity; D019098:Telomerase / Q000032:analysis / Q000378:metabolism*
D009285:Naphthoquinones; D008165:Luminol; D004247:DNA; D006861:Hydrogen Peroxide; D006735:Horseradish Peroxidase; D019098:Telomerase