Electrospin-coating of nitrocellulose membrane enhances sensitivity in nucleic acid-based lateral flow assay

2018 ◽  
Vol 1009 ◽  
pp. 81-88 ◽  
Author(s):  
Chee-Hong Takahiro Yew ◽  
Pedram Azari ◽  
Jane Ru Choi ◽  
Fei Li ◽  
Belinda Pingguan-Murphy
Keyword(s):  
Nucleic Acid ◽  
Lateral Flow ◽  
Lateral Flow Assay ◽  
Nitrocellulose Membrane

scholarly journals Glycerol Reduces Cross Hybridization on Nitrocellulose Membrane

2020 ◽  
Vol 38 (3) ◽  
pp. 199
Author(s):  
Narendra Yoga Hendarta ◽  
Abu Tholib Aman ◽  
Asmarani Kusumawati ◽  
Tri Wibawa
Keyword(s):  
Nucleic Acid ◽  
Point Of Care ◽  
Gc Content ◽  
Lateral Flow ◽  
Hybridization Signal ◽  
Nitrocellulose Membrane ◽  
High Concentration ◽  
Cross Hybridization ◽  
High Stringency ◽  
Stringency Condition

Lateral flow assay (LFD) based nucleic acid lateral flow (NALF)  method has been developed recently. The method met point of care testing (POCT) as simple and rapid procedures, less equipment, and can be performance by less skilled technician. NALF based on nucleic acid hybridizationis  more economical then immunochromatography assay which use antibody-antigen recognition. Cross hybridization has issued while used to differentiate organism with high GC content and high homology as high similarity genome. Some techniques has applied to give high stringency condition avoid cross hybridization reaction but need more procedure to apply. We found glycerol applied to buffer assay could reduce cross hybridization on nitrocellulose membrane. The study used 2 kinds of high stringency buffer as PBS and SSC bases and high concentration of ssDNA amplicon as sample. Without glycerol ingredient gave cross hybridization signal on test line. But used glycerol could reduce those even omitted with PBS based buffer assay. Beside those, glycerol could significantly increased hybridization signal in SSC based buffer assay (p<0.05).

scholarly journals One-Tube SARS-CoV-2 Detection Platform Based on RT-RPA and CRISPR/Cas12a

2020 ◽  
Author(s):  
Yangyang Sun ◽  
Lei Yu ◽  
Chengxi Liu ◽  
Wei Chen ◽  
Dechang Li ◽  
...  
Keyword(s):  
Nucleic Acid ◽  
Limit Of Detection ◽  
Lateral Flow ◽  
Lateral Flow Assay ◽  
N Gene ◽  
Clinical Samples ◽  
Nucleic Acid Detection ◽  
Detection Process ◽  
Human Coronaviruses ◽  
Negative Controls

Abstract Background: COVID-19 has spread rapidly around the world, affecting almost every person. When lifting certain mandatory measures for an economic restart, robust surveillance must be established and implemented, with nucleic acid detection for SARS-CoV-2 as an essential component. Methods: We designed RT-RPA (Reverse Transcription and Recombinase Polymerase Isothermal Amplification) primers of RdRp gene and N gene according to the SARS-CoV-2 gene sequence. We optimized the components in the reaction so that the detection process could be carried out in one tube. The specificity was demonstrated through detecting nucleic acid samples from seven human coronaviruses. Clinical samples were used to validate the platform and all results were compared to rRT-PCR. RNA standards diluted by different gradients were used to demonstrate the limit of detection. Furthermore, we have developed a lateral flow assay based on OR-DETECTR for the detection of COVID-19. Results: We have developed a o ne-tube detection platform based on R T- R PA and DNA Endonuclease-Targeted CRISPR Trans Reporter ( DETECTR ) technology, termed OR-DETECTR, to detect SARS-CoV-2. The detection process is completed in one tube, and the time is 50min. The method can specifically detect SARS-CoV-2 from seven human coronaviruses with a low detection limit of 2.5 copies/µl input. Results from six SARS-CoV-2 patient samples, eight samples from patients with fever but no SARS-CoV-2 infection, and one mixed sample from 40 negative controls showed that OR-DETECTR is 100% consistent with rRT-PCR. Furthermore, we have developed a lateral flow assay based on OR-DETECTR for the detection of COVID-19. Conclusions: OR-DETECTR detection platform is rapid, accurate, tube closed, easy-to-operate, and free of large instruments for COVID-19 detection.

scholarly journals A Novel Lateral Flow Assay for Rapid and Sensitive Nucleic Acid Detection of Avibacterium paragallinarum

2021 ◽  
Vol 8 ◽  
Author(s):  
Caiyun Huo ◽  
Donghai Li ◽  
Zhenguo Hu ◽  
Guiping Li ◽  
Yanxin Hu ◽  
...  
Keyword(s):  
Nucleic Acid ◽  
Gel Electrophoresis ◽  
Lateral Flow ◽  
Lateral Flow Assay ◽  
Nucleic Acid Hybridization ◽  
Nucleic Acid Detection ◽  
Agarose Gel ◽  
Agarose Gel Electrophoresis ◽  
Infectious Coryza ◽  
Avibacterium Paragallinarum

Avibacterium paragallinarum, the pathogen of infectious coryza, caused a highly contagious respiratory disease that poses a serious threat to chickens. Hence, it is necessary to do diagnostic screening for Av. paragallinarum. Existing technologies have been used for Av. paragallinarum testing, which, however, have some drawbacks such as time consuming and expensive that require well-trained personnel and sophisticated infrastructure, especially when they are limitedly feasible in some places for lack of resources. Nucleic acid hybridization-based lateral flow assay (LFA) is capable of dealing with these drawbacks, which is attributed to the advantages, such low cost, rapid, and simple. However, nucleic acid determination of Av. paragallinarum through LFA method has not been reported so far. In this study, we developed a novel LFA method that employed gold nanoparticle probes to detect amplified Av. paragallinarum dsDNA. Compared with agarose gel electrophoresis, this LFA strip was inexpensive, simple- to- use, and time- saving, which displayed the visual results within 5–8 min. This LFA strip had higher sensitivity that achieved the detection limit of 101 CFU/ml compared with 102 CFU/ml in agarose gel electrophoresis. Besides, great sensitivity was also shown in the LFA strip, and no cross reaction existed for other bacteria. Furthermore, Av. paragallinarum in clinical chickens with infectious coryza were perfectly detected by our established LFA strip. Our study is the first to develop the LFA integrated with amplification and sample preparation techniques for better nucleic acid detection of Av. paragallinarum, which holds great potential for rapid, accurate, and on-site determination methods for early diagnosis of Av. paragallinarum to control further spreading.

scholarly journals Improved Analytical Sensitivity of Lateral Flow Assay using Sponge for HBV Nucleic Acid Detection

2017 ◽  
Vol 7 (1) ◽  
Author(s):  
Ruihua Tang ◽  
Hui Yang ◽  
Yan Gong ◽  
Zhi Liu ◽  
XiuJun Li ◽  
...  
Keyword(s):  
Nucleic Acid ◽  
Lateral Flow ◽  
Lateral Flow Assay ◽  
Nucleic Acid Detection ◽  
Analytical Sensitivity

Direct Nucleic Acid Detection of Diarrheal Pathogens in Stool Using an Antibody-Free Lateral Flow Assay

2017 ◽  
Vol 152 (5) ◽  
pp. S817
Author(s):  
Nicolaas H. Fourie ◽  
Sarah K. Abey ◽  
Eric Ferguson ◽  
Natnael Kenea ◽  
Ana F. Diallo ◽  
...  
Keyword(s):  
Nucleic Acid ◽  
Lateral Flow ◽  
Lateral Flow Assay ◽  
Nucleic Acid Detection

scholarly journals Development of a colorimetric nucleic acid-based lateral flow assay with non-biotinylated capture DNA

2017 ◽  
Vol 60 (6) ◽  
pp. 637-645 ◽  
Author(s):  
Atefeh Javani ◽  
Fatemeh Javadi-Zarnaghi ◽  
Mohammad Javad Rasaee
Keyword(s):  
Nucleic Acid ◽  
Lateral Flow ◽  
Lateral Flow Assay

Entropy-driven amplification strategy-assisted lateral flow assay biosensor for ultrasensitive and convenient detection of nucleic acids

The Analyst ◽  
2021 ◽  
Author(s):  
Shasha Li ◽  
Hongmin Meng ◽  
Hong Zong ◽  
Juan Chen ◽  
Jianjun Li ◽  
...  
Keyword(s):  
Nucleic Acid ◽  
Nucleic Acids ◽  
Lateral Flow ◽  
Lateral Flow Assay ◽  
Treatment Measures ◽  
Amplification Strategy ◽  
And Control ◽  
Timely Treatment ◽  
Nucleic Acid Tests

Accurate, sensitive and rapid nucleic acid tests are important to implement timely treatment measures and control the spread of disease. In this work, we developed a novel portable platform for...

scholarly journals One-tube SARS-CoV-2 detection platform based on RT-RPA and CRISPR/Cas12a

2021 ◽  
Vol 19 (1) ◽  
Author(s):  
Yangyang Sun ◽  
Lei Yu ◽  
Chengxi Liu ◽  
Shanting Ye ◽  
Wei Chen ◽  
...  
Keyword(s):  
Detection Limit ◽  
Nucleic Acid ◽  
Influenza A ◽  
Lateral Flow ◽  
Lateral Flow Assay ◽  
Clinical Samples ◽  
Nucleic Acid Detection ◽  
Detection Process ◽  
Influenza A H1n1 ◽  
Human Coronaviruses

Abstract Background COVID-19 has spread rapidly around the world, affecting a large percentage of the population. When lifting certain mandatory measures for an economic restart, robust surveillance must be established and implemented, with nucleic acid detection for SARS-CoV-2 as an essential component. Methods We tried to develop a one-tube detection platform based on RT-RPA (Reverse Transcription and Recombinase Polymerase Isothermal Amplification) and DNA Endonuclease-Targeted CRISPR Trans Reporter (DETECTR) technology, termed OR-DETECTR, to detect SARS-CoV-2. We designed RT-RPA primers of the RdRp and N genes following the SARS-CoV-2 gene sequence. We optimized reaction components so that the detection process could be carried out in one tube. Specificity was demonstrated by detecting nucleic acid samples from pseudoviruses from seven human coronaviruses and Influenza A (H1N1). Clinical samples were used to validate the platform and all results were compared to rRT-PCR. RNA standards and pseudoviruses diluted by different gradients were used to demonstrate the detection limit. Additionally, we have developed a lateral flow assay based on OR-DETECTR for detecting COVID-19. Results The OR-DETECTR detection process can be completed in one tube, which takes approximately 50 min. This method can specifically detect SARS-CoV-2 from seven human coronaviruses and Influenza A (H1N1), with a low detection limit of 2.5 copies/µl input (RNA standard) and 1 copy/µl input (pseudovirus). Results of six samples from SARS-CoV-2 patients, eight samples from patients with fever but no SARS-CoV-2 infection, and one mixed sample from 40 negative controls showed that OR-DETECTR is 100% consistent with rRT-PCR. The lateral flow assay based on OR-DETECTR can be used for the detection of COVID-19, and the detection limit is 2.5 copies/µl input. Conclusions The OR-DETECTR platform for the detection of COVID-19 is rapid, accurate, tube closed, easy-to-operate, and free of large instruments.

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