high throughput detection of telomerase expression by gold nanoprobes assay (nano- trap assay): an alternative method to conventional method of telomerase detection

نویسندگان

abbas ebrahimi- kalan department of neurosciences, faculty of advanced medical sciences, tabriz university of medical sciences, tabriz, iran

shahram ahmadian clinical laboratory, imam reza hospital, tabriz university of medical sciences, tabriz, iran

hamid tayefi-nasrabadi department of anatomical, faculty of medicine, tabriz university of medical sciences, tabriz,

mohammad pourhassan-moghaddam department of medical biotechnology, faculty of advanced medical sciences, tabriz university of medical sciences, tabriz, iran c ian wark research institute, mawson lakes campus, university of south australia, mawson lakes,

چکیده

introduction: telomeric repeat amplification protocol assay (trap assay) is a standard established technique to detect the activity (expression) of telomerase in the cancerous samples. it includes a time-consuming procedure that has significant drawbacks such as high cost, complicacy, time-consuming procedure, equipment-dependent steps and false-positive signals caused by inclusion of pcr inside the assay. methods: in this paper we introduce an assay that takes the advantages of gold nanoparticles in order to avoid application of pcr and post-pcr techniques while remaining highly sensitive, specific and quantitative enough. in addition, our proposed method is much simpler, cheaper and reliable than standard trap assay. we call this new technique “nano-trap assay” as it recruits gold nanoparticles for signal amplification step. results: in nano-trap assay, telomerase elongates a suitable primer, and then gold nanoparticle-probe conjugates (gold nanoprobes) could be used to recognize the elongated products of telomerase. after that, gold or silver enhancement step could be done to translate the presence of a few gold nanoprobe-elongated primer complex to a measurable signal (signal enhancement). this enhanced signal could be inspected by naked eye, optical density measurement and analysis of their scanned images by image analysis tools. conclusion: nano-trap assay is an easy to use, low-cost and rapid alternative to conventional trap assay and since it is not very equipment demanding, it could be done in low-resources laboratories.

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