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Transitioning from Triton X-100 to Tergitol 15-S-9: impacts on diagnostic assays using viral PCR sample solution. 从 Triton X-100 过渡到 Tergitol 15-S-9:对使用病毒 PCR 样品溶液进行诊断测定的影响。
IF 2.5 4区 工程技术 Q3 BIOCHEMICAL RESEARCH METHODS Pub Date : 2024-01-01 Epub Date: 2024-05-05 DOI: 10.1080/07366205.2024.2342172
Azul Zorzoli, Alasdair MacLean, Scott Nicholson, Alison Daniels, Stephen Hughes, Susan Bennet-Slater, Christine Tait-Burkard, Noha El Sakka, Rory Gunson, Kate Templeton

In 2019, the European Union banned Triton X-100, a detergent widely used in laboratory diagnostics, including the Viral PCR Sample Solution (VPSS), and urged manufacturers to find environmentally sustainable alternatives. Tergitol 15-S-9 (VPSS2) has been proposed as an alternative surfactant. This multicenter study evaluated the effectiveness of VPSS2, a Tergitol-based viral solution, as a replacement for VPSS. Our results show the equivalent performance of VPSS2 to VPSS for nucleic acid extraction and viral stability over time at different temperatures. The new VPSS formulation was also tested against external quality assurance panels and clinical samples. The results of this work support adopting this modified viral PCR sample solution to replace Triton X-100-containing viral transport solutions.

2019 年,欧盟禁用了广泛用于实验室诊断(包括病毒 PCR 样品溶液 (VPSS))的洗涤剂 Triton X-100,并敦促制造商寻找环境可持续的替代品。Tergitol 15-S-9(VPSS2)被提议作为一种替代表面活性剂。这项多中心研究评估了基于 Tergitol 的病毒溶液 VPSS2 作为 VPSS 替代品的有效性。我们的结果表明,VPSS2 与 VPSS 在不同温度下的核酸提取和病毒稳定性方面性能相当。新的 VPSS 配方还通过了外部质量保证小组和临床样本的测试。这项工作的结果支持采用这种改良的病毒 PCR 样品溶液来替代含有 Triton X-100 的病毒运输溶液。
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引用次数: 0
Dual-mode detection of glucose based on pistol-like DNAzyme-mediated exonuclease-assisted signal cycle. 基于手枪式 DNA 酶介导的外切酶辅助信号循环的葡萄糖双模式检测。
IF 2.2 4区 工程技术 Q3 BIOCHEMICAL RESEARCH METHODS Pub Date : 2024-01-01 Epub Date: 2024-08-05 DOI: 10.1080/07366205.2024.2381403
Huiying Lu, Xiaofeng Wang

Detecting glucose accurately and sensitively from clinical samples like tears and saliva is still difficult. We have created a sensor that can detect glucose with high sensitivity and accuracy by combining the use of glucose oxidase (GOx) to catalyze glucose, a pistol-like DNAzyme (PLDz) to transform the signal, gold nanoparticles (AuNPs) to enhance the optical properties and the exonuclease-III (Exo-III) to amplify the signal. As a result, the proposed method exhibits a low detection limit of 7.5 pM and a wide detection range covering seven orders of magnitude. The suggested dual-mode strategy provides a sensitive, precise and specific detection method for glucose. Another advantage is that the dual-mode technique significantly improves the precision and consistency of the measurements, demonstrating its immense potential for use in biomedical research and clinical diagnostics.

要从泪液和唾液等临床样本中准确而灵敏地检测葡萄糖仍然很困难。我们利用葡萄糖氧化酶(GOx)催化葡萄糖、手枪样 DNA 酶(PLDz)转换信号、金纳米粒子(AuNPs)增强光学特性以及外切酶-III(Exo-III)放大信号,创造出了一种能够高灵敏度、高准确度检测葡萄糖的传感器。因此,该方法的检测限低至 7.5 pM,检测范围广达七个数量级。建议的双模式策略提供了一种灵敏、精确和特异的葡萄糖检测方法。另一个优点是,双模式技术大大提高了测量的精确度和一致性,显示了其在生物医学研究和临床诊断中的巨大应用潜力。
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引用次数: 0
The dynamic duo: cryo-EM teams up with machine learning to visualize biomolecules in motion. 动态二重奏:低温电子显微镜与机器学习相结合,实现生物分子运动的可视化。
IF 2.5 4区 工程技术 Q3 BIOCHEMICAL RESEARCH METHODS Pub Date : 2024-01-01 Epub Date: 2024-06-12 DOI: 10.1080/07366205.2024.2355771
Beatrice Bowlby

Cryo-EM has been a key technique in our understanding of biomolecular structures. Now, machine learning techniques are being used to put these structures in motion, revealing dynamic interactions and processes happening on a molecular and cellular level.[Formula: see text].

低温电子显微镜一直是我们了解生物分子结构的关键技术。现在,机器学习技术正被用于使这些结构运动起来,揭示分子和细胞水平上发生的动态相互作用和过程。
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引用次数: 0
Artificial intelligence: help or hindrance in solving the reproducibility crisis? 人工智能:帮助还是阻碍解决可重复性危机?
IF 2.5 4区 工程技术 Q3 BIOCHEMICAL RESEARCH METHODS Pub Date : 2024-01-01 Epub Date: 2024-06-20 DOI: 10.1080/07366205.2024.2355776
Jennifer Straiton

Science is in the midst of a reproducibility crisis and the integration of artificial intelligence into scientific research has only compounded the problem; yet could the technology hold the solution to its own problems?[Formula: see text].

科学正处于可重复性危机之中,人工智能融入科学研究只会使问题更加复杂;然而,这项技术能否解决自身的问题?
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引用次数: 0
Simple and sensitive detection of Pseudomonas aeruginosa in neonatal infection based on a both-end blocked peroxidase-mimicking DNAzyme. 基于双端阻断过氧化物酶模拟 DNA 酶对新生儿感染铜绿假单胞菌进行简单灵敏的检测。
IF 2.5 4区 工程技术 Q3 BIOCHEMICAL RESEARCH METHODS Pub Date : 2024-01-01 Epub Date: 2024-05-20 DOI: 10.1080/07366205.2024.2348295
Dongyun Chen, Yicong Pan, Huan Yu, Xiaoxiang Chen

Developing a simple and highly sensitive approach for Pseudomonas aeruginosa (P. aeruginosa) detection is crucial, as it is closely associated with various disorders, such as newborn infections. Nevertheless, few of techniques have the capability to accurately identify P. aeruginosa with a high level of sensitivity and significantly improved stability. The employment of the both-end blocked peroxidase-mimicking DNAzyme significantly diminished the interferences from background signals, so conferring the approach with a high degree of selectivity and reproducibility. The proposed method is demonstrated with exceptional discernment capacity in differentiating interfering microorganisms. The simplicity, elevated sensitivity and high discerning capability make the method a highly promising alternative instrument for pathogenic bacteria detection.

铜绿假单胞菌(P. aeruginosa)与新生儿感染等各种疾病密切相关,因此开发一种简单、高灵敏度的铜绿假单胞菌(P. aeruginosa)检测方法至关重要。然而,很少有技术能够准确识别铜绿假单胞菌,并具有高灵敏度和显著提高的稳定性。双端阻断过氧化物酶模拟 DNA 酶的使用大大减少了背景信号的干扰,从而使该方法具有高度的选择性和可重复性。所提出的方法在区分干扰微生物方面具有卓越的鉴别能力。该方法操作简单、灵敏度高、分辨能力强,是一种极有前途的病原菌检测替代仪器。
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引用次数: 0
Cassava for the future: embryogenic liquid cultures suitable for new biotech techniques. 未来的木薯:适合新生物技术的胚胎液体培养物。
IF 2.2 4区 工程技术 Q3 BIOCHEMICAL RESEARCH METHODS Pub Date : 2024-01-01 Epub Date: 2024-10-16 DOI: 10.1080/07366205.2024.2393546
Beata Dedičová, Luis Augusto Becerra Lopez-Lavalle

Cassava, a crop of importance for subsistence farming in Africa, Asia, and Latin America, has the potential to benefit from global economic integration as a versatile industrial resource. Enhancing cassava productivity is not just a matter of agricultural competitiveness but a crucial step toward ensuring many communities' food security and livelihoods. Given its high performance in marginal environments, where climate change poses threats, ensuring food security and livelihoods relies on rapidly adapting cassava. This study aimed to develop a protocol that swiftly transitions cassava embryogenic short-period liquid suspension cultures, facilitating the regeneration of genetically stable in vitro plants. The resulting protocol, with its potential to be a foundational component in future technologies employing various genome editing or genetic modification techniques, holds promise for the advancement of cassava biotechnology.

木薯是非洲、亚洲和拉丁美洲自给农作的重要作物,作为一种用途广泛的工业资源,它有可能从全球经济一体化中受益。提高木薯的生产率不仅关系到农业竞争力,也是确保许多社区粮食安全和生计的关键一步。鉴于木薯在气候变化构成威胁的边缘环境中表现优异,确保粮食安全和生计有赖于木薯的快速适应。本研究旨在开发一种可快速转化木薯胚胎的短周期液体悬浮培养物的方案,以促进基因稳定的体外植株的再生。由此产生的方案有可能成为未来采用各种基因组编辑或转基因技术的基础组成部分,为木薯生物技术的进步带来希望。
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引用次数: 0
Integrity of RNA in long-term-stored cervical liquid-based cytology samples: implications for biomarker research. 长期储存的宫颈液基细胞学样本中 RNA 的完整性:对生物标记研究的影响。
IF 2.5 4区 工程技术 Q3 BIOCHEMICAL RESEARCH METHODS Pub Date : 2024-01-01 Epub Date: 2024-05-01 DOI: 10.2144/btn-2023-0112
Madeleine P J White, Andrew Stevenson, Hana Elasifer, Chris Davies, Kyriaki Nomikou, Kate Cuschieri, Sheila V Graham

Biobanks of cervical screening (LBC) samples annotated with disease status are an invaluable resource to support the development of tools for the risk stratification of disease. Although there is growing interest in the assessment of RNA-based biomarkers, little is known on the suitability and durability of stored clinical samples (commonly used in cervical screening) to support RNA-based research. RNA was extracted from 260 stored LBC samples. Storage at -80°C or -25°C allowed isolation of sufficient RNA for further analysis. RNA was found to be substantially degraded according to Agilent Bioanalyser data. Despite this, RT-qPCR was successful in 95% samples tested. These data suggest that biobanked LBC samples are suitable for RNA-based assessment even if stored for up to 14 years.

注释疾病状态的宫颈筛查(LBC)样本生物库是支持疾病风险分层工具开发的宝贵资源。尽管人们对基于 RNA 的生物标记物的评估越来越感兴趣,但对储存的临床样本(常用于宫颈筛查)是否适合和耐用,是否能支持基于 RNA 的研究,却知之甚少。我们从 260 份储存的 LBC 样本中提取了 RNA。在-80°C或-25°C条件下保存,可分离出足够的RNA用于进一步分析。根据安捷伦生物分析仪的数据,发现 RNA 已严重降解。尽管如此,在 95% 的测试样本中,RT-qPCR 仍取得了成功。这些数据表明,生物库中的 LBC 样本即使保存长达 14 年,也适合进行基于 RNA 的评估。
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引用次数: 0
A standardized protocol for sample preparation for scanning electron microscopy to visualize extrachromosomal DNA. 用于扫描电子显微镜观察染色体外 DNA 的样本制备标准化方案。
IF 2.5 4区 工程技术 Q3 BIOCHEMICAL RESEARCH METHODS Pub Date : 2024-01-01 Epub Date: 2024-06-19 DOI: 10.1080/07366205.2024.2346042
Jillann A Madren, Jingting Chen, William Dennis, Christina Ford, Kristen K White, Elizabeth Brunk

Extrachromosomal DNA (ecDNA) are circular DNA structures associated with cancer and drug resistance. One specific type, double minute (DM) chromosomes, has been studied since the 1960s using imaging techniques like cytogenetics and fluorescence microscopy. Specialized techniques such as atomic force microscopy (AFM) and scanning electron microscopy (SEM) offer micro to nano-scale visualization, but current sample preparation methods may not optimally preserve ecDNA structure. Our study introduces a systematic protocol using SEM for high-resolution ecDNA visualization. We have optimized the end-to-end procedure, providing a standardized approach to explore the circular architecture of ecDNA and address the urgent need for better understanding in cancer research.

染色体外 DNA(ecDNA)是与癌症和耐药性有关的环状 DNA 结构。自 20 世纪 60 年代以来,人们一直在利用细胞遗传学和荧光显微镜等成像技术研究一种特殊类型的双微粒(DM)染色体。原子力显微镜(AFM)和扫描电子显微镜(SEM)等专业技术可提供微米到纳米尺度的可视化,但目前的样品制备方法可能无法最佳地保留 ecDNA 结构。我们的研究介绍了一种利用扫描电子显微镜进行高分辨率 ecDNA 可视化的系统方案。我们优化了端到端的程序,提供了一种标准化的方法来探索 ecDNA 的环状结构,满足了癌症研究中更好地了解其结构的迫切需要。
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引用次数: 0
Welcome to the 76th Issue of BioTechniques. 欢迎阅读第 76 期《生物技术》。
IF 2.7 4区 工程技术 Q3 BIOCHEMICAL RESEARCH METHODS Pub Date : 2024-01-01 DOI: 10.2144/btn-2023-0113
Ashling Cannon, Ebony Torrington, Tristan Free
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引用次数: 0
Novel buffer for long-term preservation of DNA in biological material at room temperature. 用于在室温下长期保存生物材料中 DNA 的新型缓冲液。
IF 2.5 4区 工程技术 Q3 BIOCHEMICAL RESEARCH METHODS Pub Date : 2024-01-01 Epub Date: 2024-07-01 DOI: 10.1080/07366205.2024.2360813
Mohaimin Kasu, Peter G Ristow, Adria Michelle Burrows, Zafrir Kuplik, Mark J Gibbons, Maria E D'Amato

The collection and preservation of biological material before DNA analysis is critical for inter alia biomedical research, medical diagnostics, forensics and biodiversity conservation. In this study, we evaluate an in-house formulated buffer called the Forensic DNA Laboratory-buffer (FDL-buffer) for preservation of biological material for long term at room temperature. Human saliva stored in the buffer for 8 years, human blood stored for 3 years and delicate animal tissues from the jellyfish Pelagia noctiluca comb jelly Beroe sp., stored for 4 and 6 years respectively consistently produced high-quality DNA. FDL-buffer exhibited compatibility with standard organic, salting out and spin-column extraction methods, making it versatile and applicable to a wide range of applications, including automation.

在 DNA 分析前收集和保存生物材料对于生物医学研究、医疗诊断、法医和生物多样性保护等至关重要。在这项研究中,我们对一种内部配制的缓冲液进行了评估,这种缓冲液被称为法医 DNA 实验室缓冲液(FDL-buffer),用于在室温下长期保存生物材料。在缓冲液中保存 8 年的人类唾液、保存 3 年的人类血液以及分别保存 4 年和 6 年的海蜇(Pelagia noctiluca comb jelly Beroe sp.FDL 缓冲液与标准的有机提取、盐析提取和旋柱提取方法兼容,因此它用途广泛,适用于包括自动化在内的各种应用。
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BioTechniques
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