文献阅读 呼吸道感染类病毒探针诊断优化课件.pptVIP

Detecting respiratory viral RNA using expanded genetic alphabets and self-avoiding DNA 季志华 2015201188 北京化工大学 生研一班 BE NOT introduction Nucleic acid (NA)-targeted tests detect and quantify viral DNA and RNA (collectively xNA) to support epidemiological surveillance and, in individual patients, to guide therapy. They commonly use polymerase chain reaction (PCR) and reverse transcription PCR. advantages: 1, Sensitivity 2, rapid disadvantages: 1,expensive Multiplexing would allow their cost to be spread over multiple targets, but often only with lower sensitivity and accuracy, noise, false positives, and false negatives. Here we offer a multiplexed assay for a panel of respiratory viruses that mitigates these problems by combining several nucleic acid analogs from the emerging field of synthetic biology. optimizing sides: (i) self-avoiding molecular recognition systems (SAMRSs), which facilitate multiplexing, (ii) artificially expanded genetic information systems(AEGISs), which enable low-noise PCR. expanded genetic information system (AEGIS) the Luminex xMAP multiplexed assays platform Materials influenzas A and B (InfA and InfB, respectively), respiratory syncytial virus (RSV), and the coronaviruses that cause severe acute respiratory syndrome (SARS) and Middle Eastrespiratory syndrome (MERS). methods 1,In vitro production of viral RNA simulants via transcription by T7 RNA polymerase. 2,viral RNA isolation 3,RTePCR 4,Digestion of excess primers and dNTPs and RPER 5,Probe coupling to Luminex MicroPlex carboxylated microspheres Results