المؤلفون: Huang, Y., Yang, J., Zeng, J., Huang, C., Gong, X., Ma, S., Ge, S., Zhang, D.

المصدر: IEEE Transactions on Instrumentation and Measurement IEEE Trans. Instrum. Meas. Instrumentation and Measurement, IEEE Transactions on. 73:1-14 2024

المؤلفون: Summa, Maija^{Aff1, IDs12560024095872_cor1}, Tuutti, Enni, Al-Hello, Haider, Huttunen, Liisa-Maija, Rimhanen-Finne, Ruska

المصدر: Food and Environmental Virology: The Official Journal of the International Society for Food and Environmental Virology. 16(2):180-187

المؤلفون: Tang, Ruihua^{Aff1, Aff2, IDs10570024058897_cor1}, Yan, Xueyan^{Aff1, Aff2}, Hu, Jie, Luo, Yining, Giesy, John P.^{Aff5, Aff6, Aff7, Aff8}, Xie, Yuwei, Yin, Huancai

المصدر: Cellulose. 31(8):5115-5131

المؤلفون: Vita Antane, Yktiyar Sarybayev, Askar Osserbay, Kudratulla Shatmanov, Tansyk Baltakhozhayev

المصدر: Наукові горизонти, Vol 27, Iss 2, Pp 31-42 (2024)

مصطلحات موضوعية: genetic material, nucleic acids, optical density of the solution, correlation, repeatability, nucleic acid extraction, Agriculture

الوصف: In animal breeding, genetic methods have become the basis of breeding work and veterinary diagnostics. Therefore, their development and improvement is an actual direction of modern science. The aim of the presented work was to study the concentration and quality of nucleic acids obtained from venous blood of cattle for further genetic studies. For this purpose, a modified method of phenolchloroform extraction, adapted for DNA extraction from blood, with subsequent spectrometric determination of DNA concentration and assessment of its quality were applied. As a result of this research, it was found that the average concentration of genetic material isolated from animal blood was 146.5±14.98 ng/µl. The main part of samples – more than 93% contained concentration of nucleic acids in the range from 50 to 200 ng/µl. At the same time, the time interval between DNA extraction and its spectrometric determination of concentration and quality of genetic material by the ratio of optical density at A260/A280 wavelengths during a year did not have significant changes on its parameters. The used method of nucleic acid extraction in 94% allowed obtaining samples of good quality suitable for further genetic studies. A correlation of 43% (P

وصف الملف: electronic resource

Relation: https://sciencehorizon.com.ua/en/journals/tom-27-2-2024/spektrofotometrichny-metod-dlya-viznachennya-kilkosti-ta-yakosti-dnk-u-tvarinnitstvi; https://doaj.org/toc/2663-2144

URL الوصول: https://doaj.org/article/98e2d4715f2d4eeda35ffa19a1aee157

المؤلفون: Huixing Ye, Wenqiang Chen, Tao Huang, Junfeng Xu, Xiaofu Wang

المصدر: Food Chemistry: Molecular Sciences, Vol 8, Iss , Pp 100206- (2024)

مصطلحات موضوعية: Nucleic acid extraction, Adulterated honey, RPA, Exogenous syrup, On-site detection, Nutrition. Foods and food supply, TX341-641

الوصف: Honey adulteration with exogenous syrup has become a common phenomenon, and current detection techniques that require large instruments are cumbersome and time-consuming. In this study, a simple and efficient method was developed by integrating the rapid extraction of nucleic acids (REMD) and recombinase polymerase amplification (RPA), known as REMD-RPA, for the rapid screening of syrup adulteration in honey. First, a rapid extraction method was developed to rapidly extract corn syrup DNA in five minutes to meet the requirements of PCR and RPA assays. Then, the RPA method for detecting endogenous maize genes (ZssIIb) was established, which could detect 12 copies/μL of the endogenous maize gene within 30 min without cross-reacting with other plant-derived genes. This indicated that the RPA technique exhibited high sensitivity and specificity. Finally, the REMD-RPA detection platform was used to detect different concentrations of corn syrup adulteration, and 1 % adulteration could be detected within 30 min. The 22 commercially available samples were tested to validate the efficacy of this method, and the established RPA was able to detect seven adulterated samples in less than 30 min. Overall, the developed method is rapid, sensitive, and specific, providing technical support for the rapid field detection of honey adulteration and can serve as a reference for developing other field test methods.

وصف الملف: electronic resource

Relation: http://www.sciencedirect.com/science/article/pii/S2666566224000133; https://doaj.org/toc/2666-5662

URL الوصول: https://doaj.org/article/112a7bbf108b4c8f912a18e121de070e

المؤلفون: Federica Valeriani, Lory Marika Margarucci, Francesca Ubaldi, Gianluca Gianfranceschi, Vincenzo Romano Spica

المصدر: Microbiology Research, Vol 15, Iss 1, Pp 120-136 (2023)

مصطلحات موضوعية: environmental surveillance, qPCR, fomites, 16S rDNA, automate nucleic acid extraction, microbiota, Microbiology, QR1-502

الوصف: During the COVID-19 pandemic, extensive efforts focused on developing a better understanding of indirect transmission routes, environmental monitoring of fomites, and suitable surveillance strategies, providing new perspectives to also face other communicable diseases. Rapid methods for monitoring environmental contamination are strongly needed to support risk assessment, epidemiological surveillance and prevent infections from spreading. We optimized and automatized a protocol based on fomite detection by qPCR, using a microbial-signature approach based on marker genes belonging to the microbiota of droplets or different biological fluids. The procedure was implemented by exploiting the available tools developed for SARS-CoV-2 tracing, such as flocked swab sampling, real-time PCR equipment and automatic extraction of nucleic acids. This approach allowed scaling up, simplifying, and speeding up the extraction step of environmental swabs, processing at least 48 samples within 45 min vs. 90 min for about 24 samples by manual protocols. A comparison of microflora data by Next-Generation Sequencing (NGS) strongly supports the effectiveness of this semiautomated extraction procedure, providing good quality DNA with comparable representation of species as shown by biodiversity indexes. Today, equipment for qPCR is widely available and relatively inexpensive; therefore this approach may represent a promising tool for hospital hygiene in surveilling fomites associated with SARS-CoV-2 or other pathogen’s transmission.

وصف الملف: electronic resource

Relation: https://www.mdpi.com/2036-7481/15/1/8; https://doaj.org/toc/2036-7481

URL الوصول: https://doaj.org/article/d3e1354b6db7400bb24cc99ff2ef7245

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المؤلفون: Chia-Wei Liu, Hideaki Tsutsui

المصدر: SLAS Technology, Vol 28, Iss 5, Pp 302-323 (2023)

مصطلحات موضوعية: Sample-to-answer, Point-of-care, Cell lysis, Nucleic acid extraction, Amplification, Biotechnology, TP248.13-248.65, Medical technology, R855-855.5

الوصف: Efficient sample preparation and accurate disease diagnosis under field conditions are of great importance for the early intervention of diseases in humans, animals, and plants. However, in-field preparation of high-quality nucleic acids from various specimens for downstream analyses, such as amplification and sequencing, is challenging. Thus, developing and adapting sample lysis and nucleic acid extraction protocols suitable for portable formats have drawn significant attention. Similarly, various nucleic acid amplification techniques and detection methods have also been explored. Combining these functions in an integrated platform has resulted in emergent sample-to-answer sensing systems that allow effective disease detection and analyses outside a laboratory. Such devices have a vast potential to improve healthcare in resource-limited settings, low-cost and distributed surveillance of diseases in food and agriculture industries, environmental monitoring, and defense against biological warfare and terrorism. This paper reviews recent advances in portable sample preparation technologies and facile detection methods that have been / or could be adopted into novel sample-to-answer devices. In addition, recent developments and challenges of commercial kits and devices targeting on-site diagnosis of various plant diseases are discussed.

وصف الملف: electronic resource

Relation: http://www.sciencedirect.com/science/article/pii/S2472630323000390; https://doaj.org/toc/2472-6303

URL الوصول: https://doaj.org/article/2d3924c1ccc8438b8859ce79c5ddf125

المؤلفون: Shuyu Lu, Yuanzhan Yang, Siqi Cui, Anyi Li, Cheng Qian, Xiaoqiong Li

المصدر: Biosensors, Vol 14, Iss 6, p 313 (2024)

مصطلحات موضوعية: centrifugal microfluidic chip, nucleic acid extraction, lamp, paraffin valve, hydrophobic valve, siphon valve, Biotechnology, TP248.13-248.65

الوصف: An integrated and high-throughput device for pathogen detection is crucial in point-of-care testing (POCT), especially for early diagnosis of infectious diseases and preventing the spread of infection. We developed an on-site testing platform that utilizes a centrifugal microfluidic chip and automated device to achieve high-throughput detection. The low-power (E. coli, our platform achieves a limit of detection (LOD) of 102 CFU/mL in 60 min. In summary, our active centrifugal microfluidic platform provides a solution for the integration of complex biological assays on turntables, with great potential in the application of point-of-care diagnosis.

وصف الملف: electronic resource

Relation: https://www.mdpi.com/2079-6374/14/6/313; https://doaj.org/toc/2079-6374

URL الوصول: https://doaj.org/article/a2a210ef074e4b539a23408e5d2feb2e

المؤلفون: Chia-Wei Liu, Sohrab Bodaghi, Georgios Vidalakis, Hideaki Tsutsui

المصدر: Chemosensors, Vol 12, Iss 6, p 105 (2024)

مصطلحات موضوعية: sample preparation, nucleic acid extraction, micro-homogenizer, paper disk, citrus diseases, Biochemistry, QD415-436

الوصف: Effective pathogen detection is essential for plant disease control. However, plant sample preparation for downstream assays, such as quantitative polymerase chain reaction (qPCR), is challenging to perform outside of a laboratory. This paper reports two sample preparation methods featuring chemical and mechanical lysis and nucleic acid extraction using a micro-homogenizer, followed by serial dilution or nucleic acid purification with a paper disk before assay. Five minutes of lysis and extraction resulted in DNA and RNA yields of up to 76.5% and 63.3%, respectively, compared to mortar and pestle controls. Crude lysates were unsuitable for direct use in qPCR assays; however, serial dilution or quick wash using chromatography paper rendered samples ready for such assays. Additionally, the nucleic acids stored on paper disks under various storage conditions remained stable for one month. These methods can facilitate the in-field preparation of citrus samples and allow for both onsite and mail-in diagnostics for growers.

وصف الملف: electronic resource

Relation: https://www.mdpi.com/2227-9040/12/6/105; https://doaj.org/toc/2227-9040

URL الوصول: https://doaj.org/article/1ce59e54b1bd4526a59eb49b7f160541