Label-Free and Sensitive Detection of BRCA1 and TB4 DNA Sequences with Water-Soluble Cationic Polythiophenes

2016 ◽  
Vol 69 (4) ◽  
pp. 473 ◽  
Author(s):  
Shaohong Zhou ◽  
Huanhuan Ling ◽  
Yun Ma ◽  
Yan Zhou ◽  
Wenqi Du ◽  
...  
Keyword(s):  
Dna Sequences ◽  
Gene Mutations ◽  
Cost Effective ◽  
Water Soluble ◽  
Label Free ◽  
Short Period ◽  
Methyl Imidazole ◽  
Volmer Constant ◽  
Initial Intensity ◽  
Breast Cancer Gene

A sensitive method for BRCA1 and TB4 DNA sequences detection using water-soluble cationic polythiophenes, poly(3-(1′-ethoxy-2′-N-methyl imidazole)thiophene) (PT) is presented. The fluorescence of PT could be dramatically quenched by the addition of single-stranded DNA (ssDNA; TB4 and BRCA1 sequences). The KSV (Stern–Volmer constant) for TB4 and BRCA1 DNA sequences are 1.46 × 108 and 3.28 × 108 M–1 respectively, and the limits of detection of these two sequences are 1.26 × 10–10 and 0.483 × 10–10 M respectively. The fluorescence of PT would recover to its initial intensity after the addition of complementary ssDNA, whereas sequences with one to three mismatched bases would not. The influences of buffer pH and concentration of NaCl were also investigated systemically in order to further improve the sensitivity. This assay can be completed in a short period of time without any further procedure. Hence, this sensitive, cost-effective, and rapid detection method for BRCA1 and TB4 DNA sequences may contribute to the clinical diagnosis of breast cancer gene mutations in the future.

Sensitive and fluorescence “turn-on” detection of BRCA1 and TB4 DNA sequences using water-soluble conjugated polythiophenes

2015 ◽  
Vol 7 (14) ◽  
pp. 5814-5819 ◽  
Author(s):  
Yun Ma ◽  
Yong Xia ◽  
Liqiang Yan ◽  
Fang Wang ◽  
Zhihui Miao ◽  
...  
Keyword(s):  
Dna Sequences ◽  
Water Soluble ◽  
Label Free ◽  
Turn On ◽  
Fluorescence Turn On

A biocompatible, label-free and sensitive fluorescence “turn-on” approach was designed to detectBRCA1andTB4DNA sequences using poly(3-[(S)-5-amino-5-carboxyl-3-oxapentyl]-2,5-thiophenylene hydrochloride) (POWT).

scholarly journals Rapid, large-scale species discovery in hyperdiverse taxa using 1D MinION sequencing

BMC Biology ◽  
2019 ◽  
Vol 17 (1) ◽  
Author(s):  
Amrita Srivathsan ◽  
Emily Hartop ◽  
Jayanthi Puniamoorthy ◽  
Wan Ting Lee ◽  
Sujatha Narayanan Kutty ◽  
...  
Keyword(s):  
Dna Sequences ◽  
Large Scale ◽  
Low Cost ◽  
Small Body ◽  
Small Subset ◽  
Similar Species ◽  
Large Species ◽  
Morphological Examination ◽  
Species Discovery ◽  
Short Period

Abstract Background More than 80% of all animal species remain unknown to science. Most of these species live in the tropics and belong to animal taxa that combine small body size with high specimen abundance and large species richness. For such clades, using morphology for species discovery is slow because large numbers of specimens must be sorted based on detailed microscopic investigations. Fortunately, species discovery could be greatly accelerated if DNA sequences could be used for sorting specimens to species. Morphological verification of such “molecular operational taxonomic units” (mOTUs) could then be based on dissection of a small subset of specimens. However, this approach requires cost-effective and low-tech DNA barcoding techniques because well-equipped, well-funded molecular laboratories are not readily available in many biodiverse countries. Results We here document how MinION sequencing can be used for large-scale species discovery in a specimen- and species-rich taxon like the hyperdiverse fly family Phoridae (Diptera). We sequenced 7059 specimens collected in a single Malaise trap in Kibale National Park, Uganda, over the short period of 8 weeks. We discovered > 650 species which exceeds the number of phorid species currently described for the entire Afrotropical region. The barcodes were obtained using an improved low-cost MinION pipeline that increased the barcoding capacity sevenfold from 500 to 3500 barcodes per flowcell. This was achieved by adopting 1D sequencing, resequencing weak amplicons on a used flowcell, and improving demultiplexing. Comparison with Illumina data revealed that the MinION barcodes were very accurate (99.99% accuracy, 0.46% Ns) and thus yielded very similar species units (match ratio 0.991). Morphological examination of 100 mOTUs also confirmed good congruence with morphology (93% of mOTUs; > 99% of specimens) and revealed that 90% of the putative species belong to the neglected, megadiverse genus Megaselia. We demonstrate for one Megaselia species how the molecular data can guide the description of a new species (Megaselia sepsioides sp. nov.). Conclusions We document that one field site in Africa can be home to an estimated 1000 species of phorids and speculate that the Afrotropical diversity could exceed 200,000 species. We furthermore conclude that low-cost MinION sequencers are very suitable for reliable, rapid, and large-scale species discovery in hyperdiverse taxa. MinION sequencing could quickly reveal the extent of the unknown diversity and is especially suitable for biodiverse countries with limited access to capital-intensive sequencing facilities.

scholarly journals Electrochemical Biosensors for Cytokine Profiling: Recent Advancements and Possibilities in the Near Future

Biosensors ◽  
2021 ◽  
Vol 11 (3) ◽  
pp. 94
Author(s):  
Nirmita Dutta ◽  
Peter B. Lillehoj ◽  
Pedro Estrela ◽  
Gorachand Dutta
Keyword(s):  
Immune System ◽  
Cost Effective ◽  
Anomalous Behavior ◽  
Quantitative Detection ◽  
Electrochemical Biosensors ◽  
Aberrant Expression ◽  
Detection Techniques ◽  
Short Period ◽  
User Friendly ◽  
Cytokine Profiling

Cytokines are soluble proteins secreted by immune cells that act as molecular messengers relaying instructions and mediating various functions performed by the cellular counterparts of the immune system, by means of a synchronized cascade of signaling pathways. Aberrant expression of cytokines can be indicative of anomalous behavior of the immunoregulatory system, as seen in various illnesses and conditions, such as cancer, autoimmunity, neurodegeneration and other physiological disorders. Cancer and autoimmune diseases are particularly adept at developing mechanisms to escape and modulate the immune system checkpoints, reflected by an altered cytokine profile. Cytokine profiling can provide valuable information for diagnosing such diseases and monitoring their progression, as well as assessing the efficacy of immunotherapeutic regiments. Toward this goal, there has been immense interest in the development of ultrasensitive quantitative detection techniques for cytokines, which involves technologies from various scientific disciplines, such as immunology, electrochemistry, photometry, nanotechnology and electronics. This review focusses on one aspect of this collective effort: electrochemical biosensors. Among the various types of biosensors available, electrochemical biosensors are one of the most reliable, user-friendly, easy to manufacture, cost-effective and versatile technologies that can yield results within a short period of time, making it extremely promising for routine clinical testing.

scholarly journals Utilising Commercially Fabricated Printed Circuit Boards as an Electrochemical Biosensing Platform

Micromachines ◽  
2021 ◽  
Vol 12 (7) ◽  
pp. 793
Author(s):  
Uroš Zupančič ◽  
Joshua Rainbow ◽  
Pedro Estrela ◽  
Despina Moschou
Keyword(s):  
Printed Circuit Boards ◽  
Electrochemical Sensors ◽  
Cost Effective ◽  
Electrochemical Sensing ◽  
Label Free ◽  
Circuit Boards ◽  
Printed Circuit ◽  
Sensing Applications ◽  
Electrochemical Biosensing ◽  
Pre Treatment

Printed circuit boards (PCBs) offer a promising platform for the development of electronics-assisted biomedical diagnostic sensors and microsystems. The long-standing industrial basis offers distinctive advantages for cost-effective, reproducible, and easily integrated sample-in-answer-out diagnostic microsystems. Nonetheless, the commercial techniques used in the fabrication of PCBs produce various contaminants potentially degrading severely their stability and repeatability in electrochemical sensing applications. Herein, we analyse for the first time such critical technological considerations, allowing the exploitation of commercial PCB platforms as reliable electrochemical sensing platforms. The presented electrochemical and physical characterisation data reveal clear evidence of both organic and inorganic sensing electrode surface contaminants, which can be removed using various pre-cleaning techniques. We demonstrate that, following such pre-treatment rules, PCB-based electrodes can be reliably fabricated for sensitive electrochemical biosensors. Herein, we demonstrate the applicability of the methodology both for labelled protein (procalcitonin) and label-free nucleic acid (E. coli-specific DNA) biomarker quantification, with observed limits of detection (LoD) of 2 pM and 110 pM, respectively. The proposed optimisation of surface pre-treatment is critical in the development of robust and sensitive PCB-based electrochemical sensors for both clinical and environmental diagnostics and monitoring applications.

Plasma enhanced label-free immunoassay for alpha-fetoprotein based on a U-bend fiber-optic LSPR biosensor

RSC Advances ◽  
2015 ◽  
Vol 5 (31) ◽  
pp. 23990-23998 ◽  
Author(s):  
Gaoling Liang ◽  
Zhongjun Zhao ◽  
Yin Wei ◽  
Kunping Liu ◽  
Wenqian Hou ◽  
...  
Keyword(s):  
Surface Plasmon Resonance ◽  
Surface Plasmon ◽  
Plasmon Resonance ◽  
Localized Surface Plasmon Resonance ◽  
Fiber Optic ◽  
Cost Effective ◽  
Alpha Fetoprotein ◽  
Localized Surface Plasmon ◽  
Label Free

A simple, label-free and cost-effective localized surface plasmon resonance (LSPR) immunosensing method was developed for detection of alpha-fetoprotein (AFP).

scholarly journals Autologous Platelet-Rich Fibrin (PRF) as an Adjunct in the Management of Osteoradionecrosis and Medication-Related Osteonecrosis of Jaws. Case Series in A Single Centre

2021 ◽  
Vol 11 (8) ◽  
pp. 3365
Author(s):  
Benjie Law ◽  
Hui Yuh Soh ◽  
Syed Nabil ◽  
Rama Krsna Rajandram ◽  
Abd Jabar Nazimi ◽  
...  
Keyword(s):  
Case Series ◽  
Cost Effective ◽  
Control Group ◽  
Platelet Rich Fibrin ◽  
Tissue Healing ◽  
Number Of Patients ◽  
Short Period ◽  
Oral And Maxillofacial Region ◽  
Surgical Adjunct ◽  
Autologous Platelet

Osteoradionecrosis (ORN) of the jaws and medication-related osteonecrosis of the jaws (MRONJ) are uncommon but serious diseases affecting the oral and maxillofacial region with clinically similar appearance but distinct pathophysiology. Management of ORN and MRONJ is inherently challenging and the treatment outcomes are unpredictable. The use of autologous platelet concentrates (APCs) to promote hard and soft tissue healing is well described in the literature, and the efficacy of leucocyte and platelet-rich fibrin (L-PRF) has been well documented in a number of clinical studies. The aim of this study was to present our treatment strategy and the outcomes of incorporating L-PRF as a surgical adjunct in management of ORN and MRONJ in our centre. Methods: eight cases of ORN and MRONJ were treated with a combination of sequestrectomy and L-PRF as a surgical adjunct. Results: the overall success was 87.5%. Using L-PRF as an adjunct, we were able to predictably manage ORN and MRONJ without causing significant morbidity. Conclusion: our experience shows that L-PRF may be used as a valuable and cost-effective adjunct to surgical management of ORN and MRONJ. However, due to a limited number of patients, and a short period of review, the true effectiveness of the method is yet to be demonstrated in a longer follow-up study including a greater number of patients, besides the inclusion of a control group.

scholarly journals Label-Free Electrochemical Biosensor Based on Au@MoS₂–PANI for Escherichia coli Detection

Chemosensors ◽  
2021 ◽  
Vol 9 (3) ◽  
pp. 49
Author(s):  
Pushap Raj ◽  
Man Hwan Oh ◽  
Kyudong Han ◽  
Tae Yoon Lee
Keyword(s):  
Escherichia Coli ◽  
Bacterial Infections ◽  
Limit Of Detection ◽  
Bacterial Pathogens ◽  
Cost Effective ◽  
Covalent Immobilization ◽  
Label Free ◽  
Self Assembled Monolayer ◽  
Detection Range ◽  
Linear Detection

Bacterial infections have become a significant challenge in terms of public health, the food industry, and the environment. Therefore, it is necessary to address these challenges by developing a rapid, cost-effective, and easy-to-use biosensor for early diagnosis of bacterial pathogens. Herein, we developed a simple, label-free, and highly sensitive immunosensor based on electrochemical detection using the Au@MoS₂–PANI nanocomposite. The conductivity of the glassy carbon electrode is greatly enhanced using the Au@MoS₂–PANI nanocomposite and a self-assembled monolayer of mercaptopropionic acid on the gold nanoparticle surface was employed for the covalent immobilization of antibodies to minimize the nonspecific adsorption of bacterial pathogens on the electrode surface. The biosensor established a high selectivity and sensitivity with a low limit of detection of 10 CFU/mL, and detected Escherichia coli within 30 min. Moreover, the developed biosensor demonstrated a good linear detection range, practical utility in urine samples, and electrode regenerative studies.

scholarly journals High-Throughput Microtiter Well-Based Chemiluminometric Genotyping of 15 HBB Gene Mutations in a Dry-Reagent Format

2007 ◽  
Vol 53 (3) ◽  
pp. 384-391 ◽  
Author(s):  
Kyriaki Glynou ◽  
Petros Kastanis ◽  
Sotiria Boukouvala ◽  
Vassilis Tsaoussis ◽  
Penelope C Ioannou ◽  
...  
Keyword(s):  
Dynamic Range ◽  
Gene Mutations ◽  
Cost Effective ◽  
Specific Probe ◽  
Duplex Pcr ◽  
Signal Ratio ◽  
Discrimination Ability ◽  
Sample Throughput ◽  
Cost Effective Method ◽  
Allele Discrimination

Abstract Background: Hemoglobinopathies are the most common inherited diseases worldwide. Various methods for genotyping of hemoglobin, beta (HBB) gene mutations have been reported, but there is need for a high sample-throughput, cost-effective method for simultaneous screening of several mutations. We report a method that combines the high detectability and dynamic range of chemiluminescence with the high allele-discrimination ability of probe extension reactions for simultaneous genotyping of 15 HBB mutations in a high sample-throughput, dry-reagent format. Methods: We genotyped the HBB mutations IVSI-110G>A, CD39C>T, IVSI-1G>A, IVSI-6T>C, IVSII-745C>G, IVSII-1G>A, FSC6GAG>G-G, −101C>T, FSC5CCT>C−, IVSI-5G>A, FSC8AAG>−G, −87C>G, IVSII-848C>A, term+6C>G, and HbS (cd6GAG>GTG). The method used comprises the following: (a) duplex PCR that produces fragments encompassing all 15 mutations, (b) probe extension reactions in the presence of fluorescein-modified dCTP, using unpurified amplicons, and (c) microtiter well-based assay of extension products with a peroxidase-antifluorescein conjugate and a chemiluminogenic substrate. We used lyophilized dry reagents to simplify the procedure and assigned the genotype by the signal ratio of the normal-to-mutant–specific probe. Results: We standardized the method by analyzing 60 samples with known genotypes and then validated by blindly genotyping 115 samples with 45 genotypes. The results were fully concordant with sequencing. The reproducibility (including PCR, probe extension reaction, and chemiluminometric assay) was studied for 20 days, and the CVs were 11%–19%. Conclusions: This method is accurate, reproducible, and cost-effective in terms of equipment and reagents. The application of the method is simple, rapid, and robust. The microtiter well format allows genotyping of a large number of samples in parallel for several mutations.

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