Direct detection of circulating microRNAs in serum of cancer patients by coupling protein-facilitated specific enrichment and rolling circle amplification

2014 ◽  
Vol 50 (25) ◽  
pp. 3292-3295 ◽  
Author(s):  
Cheng-Yi Hong ◽  
Xian Chen ◽  
Juan Li ◽  
Jing-Hua Chen ◽  
Guonan Chen ◽  
...  
Keyword(s):  
Cancer Patients ◽  
Direct Detection ◽  
Rolling Circle Amplification ◽  
Circulating Mirnas ◽  
Simple Method ◽  
Circulating Micrornas ◽  
Rolling Circle ◽  
Coupling Protein

A simple method for direct detection of circulating miRNAs in serum by coupling p19 protein-facilitated specific enrichment and RCA.

Highly Sensitive Electrochemical Biosensor for Circulating Tumor Cells Detection via Dual-Aptamer Capture and Rolling Circle Amplification Strategy

2019 ◽  
Vol 15 (7) ◽  
pp. 1568-1577 ◽  
Author(s):  
Yang Wang ◽  
Kai Chang ◽  
Cheng Yang ◽  
Shujing Li ◽  
Lixin Wang ◽  
...  
Keyword(s):  
Tumor Cells ◽  
Circulating Tumor Cells ◽  
Cancer Metastasis ◽  
Electrochemical Biosensor ◽  
Rolling Circle Amplification ◽  
K562 Cells ◽  
Detection Sensitivity ◽  
Experimental Conditions ◽  
Simple Method ◽  
Rolling Circle

A fast and simple strategy for early detection of circulating tumor cells (CTCs) is urgently required because of cancer metastasis. In this work, we assembled an electrochemical biosensor by two aptamers that could form hairpin and specifically recognize K562 cells. The thiolated capture aptamer was fixed on the gold electrode surface. The detection aptamer was linked with a primer at 3 end which could trigger rolling circle amplification to prolong the sequence of aptamer. The dual-aptamer model was fabricated to improve the capture specificity and efficiency for K562 cells. The rolling circle amplification improved the detection sensitivity by inhibiting electron transfer of [Fe(CN)6]3–/4– which could be measured by differential pulse voltammetry. The detection limit of 25 cells mL–1 and linear ranges of 1 × 10 2 to 1 × 105 cells mL–1 were obtained under optimal experimental conditions. Our work exhibited a label-free and simple method for detecting CTCs using cell-specific aptasensor, showing an expected possibility for further CTCs-related study and clinical applications of this novel method.

scholarly journals Direct detection of mRNA expression in microbial cells by fluorescence in situ hybridization using RNase H-assisted rolling circle amplification

2019 ◽  
Author(s):  
Hirokazu Takahashi ◽  
Kyohei Horio ◽  
Setsu Kato ◽  
Toshiro Kobori ◽  
Kenshi Watanabe ◽  
...  
Keyword(s):  
Mrna Expression ◽  
Direct Detection ◽  
Rolling Circle Amplification ◽  
Rnase H ◽  
Microbial Cells ◽  
Rolling Circle ◽  
The Individual ◽  
Meta Analyses

ABSTRACTMeta-analyses using next generation sequencing is a powerful strategy for studying microbiota; however, it cannot clarify the role of individual microbes within microbiota. To know which cell expresses what gene is important for elucidation of the individual cell’s function in microbiota. In this report, we developed novel fluorescence in situ hybridization (FISH) procedure using RNase-H-assisted rolling circle amplification to visualize mRNA of interest in microbial cells without reverse transcription. Our results show that this method is applicable to both gram-negative and gram-positive microbes without any noise from DNA, and it is possible to visualize the target mRNA expression directly at the single-cell level. Therefore, our procedure, when combined with data of meta-analyses, can help to understand the role of individual microbes in the microbiota.

Direct detection of circRNA in real samples using reverse transcription-rolling circle amplification

2020 ◽  
Vol 1101 ◽  
pp. 169-175 ◽  
Author(s):  
Yaqian Liu ◽  
Xiong Zhang ◽  
Min Liu ◽  
Fa Xu ◽  
Qiuyang Zhang ◽  
...  
Keyword(s):  
Reverse Transcription ◽  
Direct Detection ◽  
Rolling Circle Amplification ◽  
Rolling Circle ◽  
Real Samples

A Sensitive and Specific Hyperbranched Rolling Circle Amplification Assay and Test Strip for White Spot Syndrome Virus

2014 ◽  
Vol 97 (5) ◽  
pp. 1410-1415
Author(s):  
Yu-ran Zhao ◽  
Wei-li Yin ◽  
Zhi-qin Yue ◽  
Ba-fang Li
Keyword(s):  
White Spot Syndrome Virus ◽  
Rolling Circle Amplification ◽  
Test Strip ◽  
White Spot ◽  
Capsid Protein Gene ◽  
Simple Method ◽  
Rolling Circle ◽  
Amplification Assay ◽  
White Spot Syndrome ◽  
Global Threat

Abstract White spot syndrome virus (WSSV) is a global threat to the prawn industry, and there is no simple method for field-based testing of this virus. We designed a padlock probe and primers to the capsid protein gene VP28 of WSSV, and established a hyperbranched rolling circle amplification (HRCA) assay and a corresponding strip-based test. The assay and the test strip both had similar high accuracy and specificity, and their sensitivity was about 10 copies/μL, which is 100 times higher than conventional PCR. In this study, 68 batches of prawns were tested for WSSV with the HRCA assay and test strip, and the results were compared with the PCR assay. The results indicated that both the assay and test strip had accuracy similar to each other and to the PCR results. However, the assay and strip were more sensitive and user-friendly than PCR. Establishment of this method will provide a rapid detection of WSSV and also a basis for field-based detection of animal disease.

scholarly journals Electrochemical Genosensing of E. coli Based on Padlock Probes and Rolling Circle Amplification

Sensors ◽  
2021 ◽  
Vol 21 (5) ◽  
pp. 1749
Author(s):  
Alejandra Ben Aissa ◽  
Narayanan Madaboosi ◽  
Mats Nilsson ◽  
Maria Isabel Pividori
Keyword(s):  
Pathogenic Bacteria ◽  
Magnetic Particles ◽  
Direct Detection ◽  
Rolling Circle Amplification ◽  
Isothermal Amplification ◽  
Bacterial Dna ◽  
Rolling Circle ◽  
E Coli ◽  
Padlock Probes ◽  
Reliable Power Supply

Isothermal amplification techniques are emerging nowadays for the rapid and accurate detection of pathogenic bacteria in low resource settings, where many infectious diseases are endemic, and the lack of reliable power supply, trained personnel and specialized facilities pose critical barriers for timely diagnosis. This work addresses the detection of E. coli based on DNA isothermal amplification performed on magnetic particles (MPs) followed by electrochemical genosensing on disposable electrodes by square-wave voltammetry. In this approach, the bacterial DNA is preconcentrated using a target-specific magnetic probe and then amplified on the MPs by rolling circle amplification (RCA). Two different electrochemical readout methods for the RCA amplicons are tested. The first one relied on the labelling of the magnetic RCA product with a digoxigenin probe followed by the incubation with antiDIG-HRP antibody as electrochemical reporter. In the second case, the direct detection with an HRP-probe was performed. This latter strategy showed an improved analytical performance, while simultaneously avoiding the use of thermocyclers or bulky bench top equipment.

Rolling circle amplification and graphene-based sensor-on-a-chip for sensitive detection of serum circulating miRNAs

2019 ◽  
Vol 577 ◽  
pp. 89-97 ◽  
Author(s):  
Kiatnida Treerattrakoon ◽  
Thanakorn Jiemsakul ◽  
Chookiat Tansarawiput ◽  
Preedee Pinpradup ◽  
Tawin Iempridee ◽  
...  
Keyword(s):  
Rolling Circle Amplification ◽  
Sensitive Detection ◽  
Circulating Mirnas ◽  
Rolling Circle
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