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金獅水友至尊木蟲 (著名寫手)
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[求助]
英譯中---生物傳感器論文--求科學的翻譯
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Apart from its high sensitivity, as compared to other electrochemical miRNA biosensors, the much-simplified label- free approach makes the proposed biosensor particularly attractive for routine miRNA analysis. 30,31 To date, all miRNA assays must work with highly purified RNA samples. The freedom from chemical/biological ligation and PCR amplification greatly improves the suitability in direct profiling miRNAs with minimal or no sample pretreatment. Attempts were thus made in utilizing the biosensor for analyzing circulating miRNAs in blood and miRNAs in total RNA extract from culture cells. The same sample (except serum) was simultaneously analyzed by the biosensor and by qPCR. As presented in Table 1, good agreements between the results obtained by the biosensor and qPCR on the same sample confirmed the practical value of the biosensor. In addition, the results obtained were also consistent with published data. 32,33 Because of its high sensitivity, the amount of RNA needed for asuccessful miRNA detection was in the range of 3−10 ng. In addition, a direct analysis of let-7 miRNA in serum was attempted. Unfortunately, persistently lower values with much larger variations (30%−50%) were obtained. The results were even worse when RNA extraction efficiency is taken into consideration. Evidently, the high protein content and other blood constituents in serum severely interfere with the detection of miRNA, probably by fouling the biosensor through nonspecific adsorption. Efforts in developing an antifouling interface while maintaining most of the attractive features of the biosensor are underway. At the present time, this work only serves as a proof-of- concept through rather labor-intensive manual operation. Nonetheless, as with other electronic/electrochemical bio- sensors, both low- and high-density sensor arrays can, in principle, be constructed by adopting the standard micro- fabrication technology and the complementary metal-oxide semiconductor technology that require no additional processes other than a simple switching mechanism. Two prominent examples demonstrating the power and capacity of the electronic/electrochemical detection systems are the electro- chemical microarray developed by combimatrix 34 and the Ion Proton Sequencer recently developed by Life Technologies. 35 For example, up to 12 million individal sensors can be fabricated on a single chip in the Ion Proton Sequencer. Compared to other label-free electrochemical biosensors, 12−15 the much simplified procedure implies that the throughput can also be significantly upscaled by upgrading the manual operation to a microfluidic cartridge. Of course, similar to all other hybridization-based systems, great care must be taken when profiling multiple miRNAs sinutaneously, since there is very little room to fine-tune the hybridization conditions for all miRNAs. |
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