Pub Date : 2024-01-01Epub Date: 2024-05-05DOI: 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.
{"title":"Transitioning from Triton X-100 to Tergitol 15-S-9: impacts on diagnostic assays using viral PCR sample solution.","authors":"Azul Zorzoli, Alasdair MacLean, Scott Nicholson, Alison Daniels, Stephen Hughes, Susan Bennet-Slater, Christine Tait-Burkard, Noha El Sakka, Rory Gunson, Kate Templeton","doi":"10.1080/07366205.2024.2342172","DOIUrl":"10.1080/07366205.2024.2342172","url":null,"abstract":"<p><p>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.</p>","PeriodicalId":8945,"journal":{"name":"BioTechniques","volume":" ","pages":"295-298"},"PeriodicalIF":2.5,"publicationDate":"2024-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140847473","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"工程技术","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2024-01-01Epub Date: 2024-08-05DOI: 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。
{"title":"Dual-mode detection of glucose based on pistol-like DNAzyme-mediated exonuclease-assisted signal cycle.","authors":"Huiying Lu, Xiaofeng Wang","doi":"10.1080/07366205.2024.2381403","DOIUrl":"10.1080/07366205.2024.2381403","url":null,"abstract":"<p><p>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.</p>","PeriodicalId":8945,"journal":{"name":"BioTechniques","volume":" ","pages":"415-423"},"PeriodicalIF":2.2,"publicationDate":"2024-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141888420","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"工程技术","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2024-01-01Epub Date: 2024-06-12DOI: 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].
{"title":"The dynamic duo: cryo-EM teams up with machine learning to visualize biomolecules in motion.","authors":"Beatrice Bowlby","doi":"10.1080/07366205.2024.2355771","DOIUrl":"10.1080/07366205.2024.2355771","url":null,"abstract":"<p><p>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].</p>","PeriodicalId":8945,"journal":{"name":"BioTechniques","volume":" ","pages":"229-233"},"PeriodicalIF":2.5,"publicationDate":"2024-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141305338","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"工程技术","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2024-01-01Epub Date: 2024-06-20DOI: 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].
{"title":"Artificial intelligence: help or hindrance in solving the reproducibility crisis?","authors":"Jennifer Straiton","doi":"10.1080/07366205.2024.2355776","DOIUrl":"10.1080/07366205.2024.2355776","url":null,"abstract":"<p><p>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].</p>","PeriodicalId":8945,"journal":{"name":"BioTechniques","volume":" ","pages":"291-294"},"PeriodicalIF":2.5,"publicationDate":"2024-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141426260","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"工程技术","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2024-01-01Epub Date: 2024-05-20DOI: 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。
{"title":"Simple and sensitive detection of <i>Pseudomonas aeruginosa</i> in neonatal infection based on a both-end blocked peroxidase-mimicking DNAzyme.","authors":"Dongyun Chen, Yicong Pan, Huan Yu, Xiaoxiang Chen","doi":"10.1080/07366205.2024.2348295","DOIUrl":"10.1080/07366205.2024.2348295","url":null,"abstract":"<p><p>Developing a simple and highly sensitive approach for <i>Pseudomonas aeruginosa</i> (<i>P. aeruginosa</i>) 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 <i>P. aeruginosa</i> 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.</p>","PeriodicalId":8945,"journal":{"name":"BioTechniques","volume":"76 7","pages":"323-332"},"PeriodicalIF":2.5,"publicationDate":"2024-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142054861","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"工程技术","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2024-01-01Epub Date: 2024-10-16DOI: 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.
{"title":"Cassava for the future: embryogenic liquid cultures suitable for new biotech techniques.","authors":"Beata Dedičová, Luis Augusto Becerra Lopez-Lavalle","doi":"10.1080/07366205.2024.2393546","DOIUrl":"10.1080/07366205.2024.2393546","url":null,"abstract":"<p><p>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 <i>in vitro</i> 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.</p>","PeriodicalId":8945,"journal":{"name":"BioTechniques","volume":" ","pages":"453-461"},"PeriodicalIF":2.2,"publicationDate":"2024-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142457179","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"工程技术","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2024-01-01Epub Date: 2024-05-01DOI: 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.
{"title":"Integrity of RNA in long-term-stored cervical liquid-based cytology samples: implications for biomarker research.","authors":"Madeleine P J White, Andrew Stevenson, Hana Elasifer, Chris Davies, Kyriaki Nomikou, Kate Cuschieri, Sheila V Graham","doi":"10.2144/btn-2023-0112","DOIUrl":"10.2144/btn-2023-0112","url":null,"abstract":"<p><p>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.</p>","PeriodicalId":8945,"journal":{"name":"BioTechniques","volume":" ","pages":"245-253"},"PeriodicalIF":2.5,"publicationDate":"2024-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140852647","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"工程技术","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2024-01-01Epub Date: 2024-06-19DOI: 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.
{"title":"A standardized protocol for sample preparation for scanning electron microscopy to visualize extrachromosomal DNA.","authors":"Jillann A Madren, Jingting Chen, William Dennis, Christina Ford, Kristen K White, Elizabeth Brunk","doi":"10.1080/07366205.2024.2346042","DOIUrl":"10.1080/07366205.2024.2346042","url":null,"abstract":"<p><p>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.</p>","PeriodicalId":8945,"journal":{"name":"BioTechniques","volume":"76 7","pages":"311-321"},"PeriodicalIF":2.5,"publicationDate":"2024-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142054859","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"工程技术","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
{"title":"Welcome to the 76th Issue of <i>BioTechniques</i>.","authors":"Ashling Cannon, Ebony Torrington, Tristan Free","doi":"10.2144/btn-2023-0113","DOIUrl":"10.2144/btn-2023-0113","url":null,"abstract":"","PeriodicalId":8945,"journal":{"name":"BioTechniques","volume":"76 1","pages":"1-4"},"PeriodicalIF":2.7,"publicationDate":"2024-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"139429539","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"工程技术","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2024-01-01Epub Date: 2024-07-01DOI: 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。
{"title":"Novel buffer for long-term preservation of DNA in biological material at room temperature.","authors":"Mohaimin Kasu, Peter G Ristow, Adria Michelle Burrows, Zafrir Kuplik, Mark J Gibbons, Maria E D'Amato","doi":"10.1080/07366205.2024.2360813","DOIUrl":"10.1080/07366205.2024.2360813","url":null,"abstract":"<p><p>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 <i>Pelagia noctiluca</i> comb jelly <i>Beroe</i> 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.</p>","PeriodicalId":8945,"journal":{"name":"BioTechniques","volume":" ","pages":"357-370"},"PeriodicalIF":2.5,"publicationDate":"2024-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141466005","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"工程技术","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}