scholarly journals Highly sensitive and multiplexed one-step RT-qPCR for profiling genes involved in the circadian rhythm using microparticles

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Mi Yeon Kim ◽  
Seungwon Jung ◽  
Junsun Kim ◽  
Heon Jeong Lee ◽  
Seunghwa Jeong ◽  
...  

AbstractGiven the growing interest in molecular diagnosis, highly extensive and selective detection of genetic targets from a very limited amount of samples is in high demand. We demonstrated the highly sensitive and multiplexed one-step RT-qPCR platform for RNA analysis using microparticles as individual reactors. Those particles are equipped with a controlled release system of thermo-responsive materials, and are able to capture RNA targets inside. The particle-based assay can successfully quantify multiple target RNAs from only 200 pg of total RNA. The assay can also quantify target RNAs from a single cell with the aid of a pre-concentration process. We carried out 8-plex one-step RT-qPCR using tens of microparticles, which allowed extensive mRNA profiling. The circadian cycles were shown by the multiplex one-step RT-qPCR in human cell and human hair follicles. Reliable 24-plex one-step RT-qPCR was developed using a single operation in a PCR chip without any loss of performance (i.e., selectivity and sensitivity), even from a single hair. Many other disease-related transcripts can be monitored using this versatile platform. It can also be used non–invasively for samples obtained in clinics.

RSC Advances ◽  
2021 ◽  
Vol 11 (34) ◽  
pp. 20720-20724
Author(s):  
Guangyu Zhu ◽  
Hanjia Hu ◽  
Tao Yang ◽  
Junjun Ma ◽  
Sanjun Zhang ◽  
...  

Silver nanoclusters were synthesized through one-step UV photoreduction using the multiple carboxyl copolymer poly(methacrylic acid-co-itaconic acid) as a novel template, which can be used for highly sensitive and selective detection of Cu2+ by fluorescence quenching.


2018 ◽  
Vol 42 (4) ◽  
pp. 3097-3101 ◽  
Author(s):  
Qiaoling Liu ◽  
Nan Zhang ◽  
Haoying Shi ◽  
Wenyan Ji ◽  
Xiaoqian Guo ◽  
...  

Schematic illustration of the synthesis of CDs and the recognition process for Cu2+.


2019 ◽  
Author(s):  
Ayumu Karimata ◽  
Pradnya Patil ◽  
Eugene Khaskin ◽  
Sébastien Lapointe ◽  
robert fayzullin ◽  
...  

Direct translation of mechanical force into changes in chemical behavior on a molecular level has important implication not only for the fundamental understanding of mechanochemical processes, but also for the development of new stimuli-responsive materials. In particular, detection of mechanical stress in polymers via non-destructive methods is important in order to prevent material failure and to study the mechanical properties of soft matter. Herein, we report that highly sensitive changes in photoluminescence intensity can be observed in response to the mechanical stretching of cross-linked polymer films when using stable, (pyridinophane)Cu-based dynamic mechanophores. Upon stretching, the luminescence intensity increases in a fast and reversible manner even at small strain (< 50%) and applied stress (< 0.1 MPa) values. Such sensitivity is unprecedented when compared to previously reported systems based on organic mechanophores. The system also allows for the detection of weak mechanical stress by spectroscopic measurements or by direct visual methods.<br>


2021 ◽  
Vol 233 ◽  
pp. 117911
Author(s):  
Minmin Wang ◽  
Linxia Lu ◽  
Wenwu Song ◽  
Xunyue Wang ◽  
Tongming Sun ◽  
...  

2020 ◽  
Vol 17 (1) ◽  
Author(s):  
Yang Zhang ◽  
Chunyang Dai ◽  
Huiyan Wang ◽  
Yong Gao ◽  
Tuantuan Li ◽  
...  

Abstract Background Coronavirus disease 2019 (COVID-19), caused by SARS-CoV-2, is posing a serious threat to global public health. Reverse transcriptase real-time quantitative polymerase chain reaction (qRT-PCR) is widely used as the gold standard for clinical detection of SARS-CoV-2. Due to technical limitations, the reported positive rates of qRT-PCR assay of throat swab samples vary from 30 to 60%. Therefore, the evaluation of alternative strategies to overcome the limitations of qRT-PCR is required. A previous study reported that one-step nested (OSN)-qRT-PCR revealed better suitability for detecting SARS-CoV-2. However, information on the analytical performance of OSN-qRT-PCR is insufficient. Method In this study, we aimed to analyze OSN-qRT-PCR by comparing it with droplet digital PCR (ddPCR) and qRT-PCR by using a dilution series of SARS-CoV-2 pseudoviral RNA and a quality assessment panel. The clinical performance of OSN-qRT-PCR was also validated and compared with ddPCR and qRT-PCR using specimens from COVID-19 patients. Result The limit of detection (copies/ml) of qRT-PCR, ddPCR, and OSN-qRT-PCR were 520.1 (95% CI: 363.23–1145.69) for ORF1ab and 528.1 (95% CI: 347.7–1248.7) for N, 401.8 (95% CI: 284.8–938.3) for ORF1ab and 336.8 (95% CI: 244.6–792.5) for N, and 194.74 (95% CI: 139.7–430.9) for ORF1ab and 189.1 (95% CI: 130.9–433.9) for N, respectively. Of the 34 clinical samples from COVID-19 patients, the positive rates of OSN-qRT-PCR, ddPCR, and qRT-PCR were 82.35% (28/34), 67.65% (23/34), and 58.82% (20/34), respectively. Conclusion In conclusion, the highly sensitive and specific OSN-qRT-PCR assay is superior to ddPCR and qRT-PCR assays, showing great potential as a technique for detection of SARS-CoV-2 in patients with low viral loads.


RSC Advances ◽  
2021 ◽  
Vol 11 (24) ◽  
pp. 14700-14709
Author(s):  
Rintumoni Paw ◽  
Moushumi Hazarika ◽  
Purna K. Boruah ◽  
Amlan Jyoti Kalita ◽  
Ankur K. Guha ◽  
...  

Synthesis of Ag nanoparticles using Allin based garlic extract for highly sensitive and selective detection of metal ions Hg2+ and Sn2+ in water. The limit of detection (LoD) for Hg2+ and Sn2+ ions were found as 15.7 nM and 11.25 nM respectively.


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