TALANTA
·
2023
·
IF 6.1
Haodong Cui,Wenxing Song,Xiaoling Ru,Wen Fu,Ling Ji,Wenhua Zhou,Zhen Zhao,Guangbo Qu,Xue-Feng Yu,Guibin Jiang
The ongoing outbreak of the novel coronavirus disease 2019 (COVID-19) draws worldwide concerns due to its long incubation period and strong infectivity. Although RT-PCR-based methods are being widely applied for clinical diagnosis, timely and accurate diagnosis towards COVID-19 causing virus, the SARS-CoV-2, is still limited due to labor-intensive and time-consuming operations. Herein, we report a new viral RNA extraction method based on poly-(amino ester) with carboxyl group (PC)-coated magnetic nanoparticles (pcMNPs) for the sensitive detection of SARS-CoV-2. This method combines the lysis and binding steps into one step, and refines multiple washing steps into one step, giving a turnaround time of less than 9 min. Furthermore, the extracted pcMNP-RNA complexes can be directly introduced into subsequent RT-PCR reactions without elution. This simplified viral RNA method could be well adapted in fast manual and automated high-throughput nucleic acids extraction protocols suitable for different scenarios. A high sensitivity down to 100 copies/mL and a linear correlation between 100 and 10 6 copies/mL of SARS-CoV-2 pseudovirus particles are achieved in both protocols. Benefitting from the simplicity and excellent performances, this new method can dramatically improve the efficiency and reduce operational requirements for the early clinical diagnosis and large-scale SARS-CoV-2 nucleic acid screening.
研究领域:
分子生物学
·
医学诊断
·
纳米技术
·
传染病研究
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产品关联
DOI
PubMed
高影响因子
LANGMUIR
·
2022
·
IF 4.331
Feixia Peng,Xuelian Wang,Wenting Tao,Yao Chen,Yani Ma,Xueqin Ding
Four kinds of hydrophobic magnetic deep eutectic solvents (HMDESs) were prepared and applied to RNA extraction. Based on the HMDESs, a mechanical shaking-assisted liquid–liquid extraction (MSLLE) was developed for the extraction of RNA. Factors that influence the extraction, including the extraction time, temperature, volume of HMDES, buffer types, and pH, were evaluated. After the optimization of all conditions, the RNA extraction efficiency was 82.31 ± 0.02%. RNA can be extracted from complex samples and medicinal yeast by the method proposed in this work and can be recovered from the HMDESs after being extracted.
研究领域:
生物化学
·
分子生物学
✓ 已验证
DOI
PubMed
高影响因子
ACS Applied Materials & Interfaces
·
2023
·
IF 9.5
Zhongjie Fei,Ping Liu,Chu Cheng,Rongbin Wei,Pengfeng Xiao,Youfa Zhang
Although numerous approaches were proposed for the nucleic acid (NA)-based SARS-CoV-2 detection, the nonideal NA desorption efficiency of conventional magnetic beads (MBs) limits their widespread application. In this study, we developed solvent-responsive MBs (called responsive MBs), which, in the presence of buffers, modulated the absorption and desorption capacities of NA by flipping the surface −COO–. Relative to other commercial MBs, responsive MBs exhibited similar absorption profiles and markedly enhanced desorption profiles. When applied for NA detection of complex samples, responsive MBs exhibited better performance of RNA detection than DNA, with obvious advantages in sensitivity. Specifically, the RNA and DNA desorption rates of commercial MBs were ∼85 and 82.5%, while those of responsive MBs were nearly 94 and 93.5%, respectively. Furthermore, responsive MBs exhibited remarkable extraction ability in a wide range of tissues and better performance of RNA extraction than DNA. When applied for SARS-CoV-2 detection, the responsive MBs along with the simulated digital RT-LAMP (a previously established apparatus) further improved detection efficiency, yielding a precise quantitative detection as low as 25 copies and an ultimate sensibility detection of 5 copies/mL. It was also successfully employed in numerous NA-based technologies such as polymerase chain reaction (PCR), sequencing, and so on.
研究领域:
生物技术
·
分子生物学
·
病毒学
·
分析化学
✓ 已验证
产品关联
DOI
PubMed
高影响因子
LANGMUIR
·
2022
·
IF 4.331
Yani Ma,Yao Chen,Feixia Peng,Xueqin Ding
A phenylpropyl guanidinium magnetic ionic liquid (PGMIL) was designed and prepared to extract RNA from complex samples. The properties of PGMIL were characterized by a vibrating sample magnetometer, Fourier transform infrared spectrometer, thermogravimetric analyzer, transmission electron microscope, and scanning electron microscope. Through single-factor analysis, the factors affecting the RNA extraction process, such as PGMIL volume, temperature, extraction time, and pH, were systematically investigated. The ability of PGMIL to selectively extract RNA was investigated by a NanoDrop. Under the optimized conditions, the extraction efficiency of RNA can reach 81.9 ± 1.9%. The proposed extraction method has been demonstrated with the extraction of RNA from a series of complex sample matrices, including a metal ion mixture and medicinal yeast. After extraction, the retained RNA could be readily recovered by simply using Tris–HCl buffer, with a recovery rate of 68.11 ± 2.45%. Regeneration studies have shown that the extraction efficiency of PGMIL did not change significantly after using 4 times. This study provides a green, rapid, and environmental friendly extraction method for the selective extraction of RNA.
研究领域:
生物化学
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分析化学
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产品关联
DOI
PubMed
高影响因子
ANALYST
·
2012
·Vol. 137
·期 6
·1389-1395
·
IF 4.23
Huanshun Yin,Yunlei Zhou,Chuanxia Chen,Lusheng Zhu,Shiyun Ai
The abnormal expression of microRNAs (miRNAs) in many solid tumors makes miRNAs potential biomarkers for disease diagnosis and highlights the need for the sensitive and selective detection of miRNAs. In the present work, an ‘off-on’ signaling genosensor platform for miRNA-21 detection was well developed. This tactic was based on a locked nucleic acid-integrated nucleic acid hairpin probe, a biotin-labeled bridge DNA–AuNPs–bio-barcode signal amplification unit and enzymatic signal amplification. The test is simple, fast and ultrasensitive with a linear range of 0.01–700 pM. The detection limit was estimated to be 6 fM. The overexpression of miRNA-21 was confirmed in total RNA extracted from human hepatocarcinoma cells BEL-7402 and human HeLa cells compared with the control sample extracted from normal human hepatic L02 cells. This method does not need miRNA-21 labeling, isolation, enrichment or PCR amplification. The performance of the assay developed here could satisfy the need for rapid, easy, sensitive and specific early cancer diagnosis in clinical diagnostics.
研究领域:
生物化学
·
分子生物学
·
癌症研究
·
生物技术
✓ 已验证
产品关联
DOI
PubMed
高影响因子
危险品化学品经营许可证（带存储）