scholarly journals A Novel Design Combining Isothermal Exponential Amplification and Gold-Nanoparticles Visualization for Rapid Detection of miRNAs

2018 ◽  
Vol 19 (11) ◽  
pp. 3374 ◽  
Author(s):  
Jiquan Jiang ◽  
Bin Zhang ◽  
Chi Zhang ◽  
Yifu Guan

MicroRNAs (miRNAs) play important roles in a wide range of biological processes, and their aberrant expressions are associated with various diseases. The levels of miRNAs can be useful biomarkers for cellular events or disease diagnosis; thus, sensitive and selective detection of microRNAs is of great significance in understanding biological functions of miRNAs, early-phase diagnosis of cancers, and discovery of new targets for drugs. However, traditional approaches for the detection of miRNAs are usually laborious and time-consuming, with a low sensitivity. Here, we develop a simple, rapid, ultrasensitive colorimetric assay based on the combination of isothermal Exponential Amplification Reaction (EXPAR) and AuNP-labeled DNA probes for the detection of miRNAs (taking let-7a as a model analyte). In this assay, the presence of let-7a is converted to the reporter Y through EXPAR under isothermal conditions. The subsequent sandwich hybridization of the reporter Y with the AuNP-labeled DNA probes generates a red-to-purple color change. In other words, if the reporter Y is complementary to the AuNP-labeled DNA probes, the DNA-functionalized AuNPs will be aggregated, resulting in the change of solution color from red to purple/blue, while when the AuNP-labeled DNA probes are mismatched to the reporter Y, the solution remains red. This assay represents a simple, time-saving technique, and its results can be visually detected with the naked eye due to the colorimetric change. The method provides superior sensitivity, with a detection limit of 4.176 aM over a wide range from 1 nM to 1 aM under optimal conditions. The method also shows high selectivity for discriminating even single-nucleotide differences between let-7 miRNA family members. Notably, it is comparable to the most sensitive method reported to date, thus providing a promising alternative to standard approaches for the direct detection of let-7a miRNA. Importantly, through combination with specific templates, different miRNAs can be converted to the same reporter Y, which can hybridize with the same set of AuNP-labeled DNA probes to form sandwich hybrids. The color change of the solution can be observed in the presence of the target miRNA. This technique has potential as a routine method for assessing the levels of miRNAs, not only for let-7, but also for various miRNAs in the early phase of cancers. In addition, it can be a useful tool in biomedical research and clinical diagnosis, as well as diagnosis or surveillance programs in field conditions.

2019 ◽  
pp. 28-34
Author(s):  
Margarita Castillo-Téllez ◽  
Beatriz Castillo-Téllez ◽  
Juan Carlos Ovando-Sierra ◽  
Luz María Hernández-Cruz

For millennia, humans have used hundreds of medicinal plants to treat diseases. Currently, many species with important characteristics are known to alleviate a wide range of health problems, mainly in rural areas, where the use of these resources is very high, even replacing scientific medicine almost completely. This paper presents the dehydration of medicinal plants that are grown in the State of Campeche through direct and indirect solar technologies in order to evaluate the influence of air flow and temperature on the color of the final product through the L* a* scale. b*, analyzing the activity of water and humidity during the drying process. The experimental results showed that the direct solar dryer with forced convection presents a little significant color change in a drying time of 400 min on average, guaranteeing the null bacterial proliferation and reaching a final humidity between 9 % and 11 %.


Micromachines ◽  
2020 ◽  
Vol 11 (1) ◽  
pp. 72 ◽  
Author(s):  
Da-Quan Yang ◽  
Bing Duan ◽  
Xiao Liu ◽  
Ai-Qiang Wang ◽  
Xiao-Gang Li ◽  
...  

The ability to detect nanoscale objects is particular crucial for a wide range of applications, such as environmental protection, early-stage disease diagnosis and drug discovery. Photonic crystal nanobeam cavity (PCNC) sensors have attracted great attention due to high-quality factors and small-mode volumes (Q/V) and good on-chip integrability with optical waveguides/circuits. In this review, we focus on nanoscale optical sensing based on PCNC sensors, including ultrahigh figure of merit (FOM) sensing, single nanoparticle trapping, label-free molecule detection and an integrated sensor array for multiplexed sensing. We believe that the PCNC sensors featuring ultracompact footprint, high monolithic integration capability, fast response and ultrahigh sensitivity sensing ability, etc., will provide a promising platform for further developing lab-on-a-chip devices for biosensing and other functionalities.


Sensors ◽  
2021 ◽  
Vol 21 (4) ◽  
pp. 1109
Author(s):  
Varnakavi. Naresh ◽  
Nohyun Lee

A biosensor is an integrated receptor-transducer device, which can convert a biological response into an electrical signal. The design and development of biosensors have taken a center stage for researchers or scientists in the recent decade owing to the wide range of biosensor applications, such as health care and disease diagnosis, environmental monitoring, water and food quality monitoring, and drug delivery. The main challenges involved in the biosensor progress are (i) the efficient capturing of biorecognition signals and the transformation of these signals into electrochemical, electrical, optical, gravimetric, or acoustic signals (transduction process), (ii) enhancing transducer performance i.e., increasing sensitivity, shorter response time, reproducibility, and low detection limits even to detect individual molecules, and (iii) miniaturization of the biosensing devices using micro-and nano-fabrication technologies. Those challenges can be met through the integration of sensing technology with nanomaterials, which range from zero- to three-dimensional, possessing a high surface-to-volume ratio, good conductivities, shock-bearing abilities, and color tunability. Nanomaterials (NMs) employed in the fabrication and nanobiosensors include nanoparticles (NPs) (high stability and high carrier capacity), nanowires (NWs) and nanorods (NRs) (capable of high detection sensitivity), carbon nanotubes (CNTs) (large surface area, high electrical and thermal conductivity), and quantum dots (QDs) (color tunability). Furthermore, these nanomaterials can themselves act as transduction elements. This review summarizes the evolution of biosensors, the types of biosensors based on their receptors, transducers, and modern approaches employed in biosensors using nanomaterials such as NPs (e.g., noble metal NPs and metal oxide NPs), NWs, NRs, CNTs, QDs, and dendrimers and their recent advancement in biosensing technology with the expansion of nanotechnology.


2021 ◽  
pp. 104063872110245
Author(s):  
Sabri A. Rahman ◽  
Kuan H. Khor ◽  
Siti Khairani-Bejo ◽  
Seng F. Lau ◽  
Mazlina Mazlan ◽  
...  

Leptospirosis is a serious bacterial disease that affects both humans and animals. A wide range of symptoms have been described in humans; the disease in dogs is commonly associated with kidney and/or liver disease. In Malaysia, information about the common serovars infecting dogs is limited. Therefore, we investigated the occurrences of leptospirosis in 124 pet dogs diagnosed with kidney and/or liver disease. Blood, urine, abdominal effusion, and/or kidney and liver were collected from the dogs. Based on microscopic agglutination testing, 53 of 124 (42.7%) dogs were seropositive for leptospiral exposure. Sera were frequently positive to serovars Bataviae ( n = 12), Javanica ( n = 10), and Icterohaemorrhagiae ( n = 10). Direct detection using PCR showed that 42 of 124 (33.9%) of the whole blood and 36 of 113 (31.9%) urine samples were positive for pathogenic Leptospira spp. By PCR, 2 of 23 (9.1%) kidney and 2 of 23 (9.1%) liver were positive for pathogenic Leptospira spp. Abdominal effusion from 4 dogs were PCR-positive for pathogenic Leptospira spp. The species detected were L. interrogans, L. borgpetersenii, L. kirschneri, and L. kmetyi by partial 16S rRNA sequencing. We further identified and characterized 11 Leptospira spp. isolates from 8 dogs as serovars Bataviae, Javanica, and Australis. The mortality rate of the Leptospira-infected dogs was high (18 of 53; 34%).


2021 ◽  
Vol 181 ◽  
pp. 113118
Author(s):  
Seongjae Jo ◽  
Wonseok Lee ◽  
Joohyung Park ◽  
Hyunjun Park ◽  
Minwoo Kim ◽  
...  

2021 ◽  
Vol 143 (3) ◽  
Author(s):  
Suhui Li ◽  
Huaxin Zhu ◽  
Min Zhu ◽  
Gang Zhao ◽  
Xiaofeng Wei

Abstract Conventional physics-based or experimental-based approaches for gas turbine combustion tuning are time consuming and cost intensive. Recent advances in data analytics provide an alternative method. In this paper, we present a cross-disciplinary study on the combustion tuning of an F-class gas turbine that combines machine learning with physics understanding. An artificial-neural-network-based (ANN) model is developed to predict the combustion performance (outputs), including NOx emissions, combustion dynamics, combustor vibrational acceleration, and turbine exhaust temperature. The inputs of the ANN model are identified by analyzing the key operating variables that impact the combustion performance, such as the pilot and the premixed fuel flow, and the inlet guide vane angle. The ANN model is trained by field data from an F-class gas turbine power plant. The trained model is able to describe the combustion performance at an acceptable accuracy in a wide range of operating conditions. In combination with the genetic algorithm, the model is applied to optimize the combustion performance of the gas turbine. Results demonstrate that the data-driven method offers a promising alternative for combustion tuning at a low cost and fast turn-around.


Micromachines ◽  
2018 ◽  
Vol 9 (10) ◽  
pp. 502 ◽  
Author(s):  
Christopher Lim ◽  
Yangchung Lee ◽  
Lawrence Kulinsky

We report on the fabrication of a syringe-based platform for automation of a colorimetric malaria-Ab assay. We assembled this platform from inexpensive disposable plastic syringes, plastic tubing, easily-obtainable servomotors, and an Arduino microcontroller chip, which allowed for system automation. The automated system can also be fabricated using stereolithography (SLA) to print elastomeric reservoirs (used instead of syringes), while platform framework, including rack and gears, can be printed with fused deposition modeling (FDM). We report on the optimization of FDM and SLA print parameters, as well as post-production processes. A malaria-Ab colorimetric test was successfully run on the automated platform, with most of the assay reagents dispensed from syringes. Wash solution was dispensed from an SLA-printed elastomeric reservoir to demonstrate the feasibility of both syringe and elastomeric reservoir-based approaches. We tested the platform using a commercially available malaria-Ab colorimetric assay originally designed for spectroscopic plate readers. Unaided visual inspection of the assay solution color change was sufficient for qualitative detection of positive and negative samples. A smart phone application can also be used for quantitative measurement of the assay color change.


2021 ◽  
Author(s):  
Deepshikha Shahdeo ◽  
Azmat Ali Khan ◽  
Amer M Alanazi ◽  
Yun Suk Huh ◽  
Shruti Shukla ◽  
...  

Abstract Ochratoxin A (OTA) is one of the predominant mycotoxins that contaminate a wide range of food commodities. In the present study, a 36-mer aptamer was used as a molecular recognition element coupled with gold nanoparticles (AuNPs) for colorimetric detection of OTA in a microfluidic paper-based analytical device (µPADs). The µPADs consisted of three zones: control, detection, and sample, interconnected by channels. The biophysical characterizations of aptamer conjugated AuNPs were done by UV-vis spectroscopy (UV-vis), dynamic Light Scattering (DLS), and transmission electron microscopy (TEM). The developed colorimetric assay for OTA showed a limit of detection of 242, 545, and 95.69 ng/mL in water, corn, and groundnut, respectively. The HPLC detection method achieved acceptable coefficient in standard curves (r2 = 0.9995), better detection range, and recovery rates in spiked corn and groundnut samples as 43.61 ± 2.18% to 87.10 ± 1.82% and 42.01 ± 1.31% to 86.03 ± 2.64% after multiple sample extractions and cleanup steps. However, the developed µPADs analytical device had the potent ability to rapidly detect OTA without any extraction pre-requirement, derivatization, and cleanup steps, thus illustrating its feasibility in the animal health sector, agricultural, and food industries.


2008 ◽  
Vol 54 (9) ◽  
pp. 742-747 ◽  
Author(s):  
Shiyong Lin ◽  
Xinying Wang ◽  
Haoxuan Zheng ◽  
Zhengguo Mao ◽  
Yong Sun ◽  
...  

Our purpose was to establish a quick and accurate real-time PCR (rtPCR) method to detect Campylobacter jejuni directly from human diarrheal stool as an alternative to traditional culture methods. To determine the consistency of rtPCR and culture method, 256 clinical diarrheal stool samples and 50 normal stool samples from healthy individuals were examined, and the whole process was double-blinded. Our data showed that the sensitivity of rtPCR in pure cultures and stool was 102CFU·mL–1and 103CFU·g–1, respectively. Of the 256 diarrheal samples, 10 specimens were successfully detected by both methods, whereas two specimens were PCR positive but culture negative. No positive results were found by these two methods in 50 normal specimens. Our data suggested that rtPCR was convenient in operation and time-saving (turnaround time 3.5–4 h), so it could be used for clinical diagnostic and epidemiological purposes.


1980 ◽  
Vol 28 (2) ◽  
pp. 381-386
Author(s):  
C Sadarangani ◽  
E Skamene ◽  
P A Kongshavn

The characteristics of the mononuclear phagocytes mediating resistance to infection with Listeria during the early phase (0 to 48 h) of the response have been investigated in genetically determined susceptible (A/J) and resistant (C57BL/6, B10.A/SgSn) strains of mice. Irradiation immediately before infection profoundly enhanced the bacterial growth in the resistant strain, while having no effect in the susceptible strain, over a wide range (3 x 10(3) to 10(5)) of infective doses. This effect of irradiation is demonstrable at low-dose radiation (200 roentgens) and can be reversed by repopulation with 20 x 10(6) syngeneic nucleated bone marrow cells. Administration of dextran sulfate 500 24 h before infection profoundly enhanced the bacterial growth in the susceptible strain, while having much less effect in the resistant strain. Thus, the genetic advantage of the resistant mouse strains to listerial infection, at least during the early phase of the response, appears to be due to a cellular mechanism that is highly radiosensitive and relatively insensitive to dextran sulfate 500. In the susceptible strain, the early protective cellular mechanism is radioresistant and highly dextran sulfate 500 sensitive.


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