A novel method for room temperature distribution and conservation of RNA and DNA reference materials for guaranteeing performance of molecular diagnostics in onco-hematology: A GBMHM study

Cayuela, Jean‐Michel; Mauté, Carole; Fabre, Anne-Lise; Nibourel, Olivier; Dulucq, Stéphanie; Delabesse, Éric; Villarèse, Patrick; Hayette, Sandrine; Mozziconacci, Marie‐Joëlle; Macintyre, Elizabeth

doi:10.1016/j.clinbiochem.2015.04.004

Cited by 5 publications

(6 citation statements)

References 16 publications

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“…The mid- and long-term stability of the quality of the samples conserved at low temperature is often difficult to maintain and control, requiring substantial precautions [ 52 , 53 , 54 ]. It is for this reason that several procedures have been developed to maintain the quality and stability at RT of nucleic acids over a long period of time for subsequent use in research [ 55 , 56 , 57 , 58 , 59 , 60 , 61 ].…”

Section: Discussionmentioning

confidence: 99%

“…An arbitrary number of eight capsules per sample was chosen, each containing a minimum of 400 ng of DNA. Encapsulation was performed as described previously [ 55 , 56 , 68 , 81 ]. Briefly, aliquots of the samples were made using a specifically configured instrument to avoid pipetting errors and contamination.…”

Section: Methodsmentioning

confidence: 99%

“…Briefly, aliquots of the samples were made using a specifically configured instrument to avoid pipetting errors and contamination. The DNA was put into stainless steel capsules and subjected to lyophilization [ 55 , 56 , 68 , 81 ]. The capsules were labeled with a 2D code with a laser to assure tracking.…”

Section: Methodsmentioning

confidence: 99%

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Ensuring the Safety and Security of Frozen Lung Cancer Tissue Collections through the Encapsulation of Dried DNA

Washetine

Kara-Borni

Heeke

et al. 2018

Cancers

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Collected specimens for research purposes may or may not be made available depending on their scarcity and/or on the project needs. Their protection against degradation or in the event of an incident is pivotal. Duplication and storage on a different site is the best way to assure their sustainability. The conservation of samples at room temperature (RT) by duplication can facilitate their protection. We describe a security system for the collection of non-small cell lung cancers (NSCLC) stored in the biobank of the Nice Hospital Center, France, by duplication and conservation of lyophilized (dried), encapsulated DNA kept at RT. Therefore, three frozen tissue collections from non-smoking, early stage and sarcomatoid carcinoma NSCLC patients were selected for this study. DNA was extracted, lyophilized and encapsulated at RT under anoxic conditions using the DNAshell technology. In total, 1974 samples from 987 patients were encapsulated. Six and two capsules from each sample were stored in the biobanks of the Nice and Grenoble (France) Hospitals, respectively. In conclusion, DNA maintained at RT allows for the conservation, duplication and durability of collections of interest stored in biobanks. This is a low-cost and safe technology that requires a limited amount of space and has a low environmental impact.

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Section: Discussionmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

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Ensuring the Safety and Security of Frozen Lung Cancer Tissue Collections through the Encapsulation of Dried DNA

Washetine

Kara-Borni

Heeke

et al. 2018

Cancers

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“…In this context, an innovative procedure, where desiccated purified DNA or RNA, are maintained under an anoxic and anhydrous atmosphere inside hermetic minicapsules has been developed by Imagene [ 29 , 30 , 31 ] and independently validated [ 32 , 33 ]. This process allows a standalone storage (no need for humidity or temperature controls) and enables long term stability of DNA at ambient temperature (over centuries).…”

Section: Introductionmentioning

confidence: 99%

High DNA stability in white blood cells and buffy coat lysates stored at ambient temperature under anoxic and anhydrous atmosphere

et al. 2017

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Conventional storage of blood-derived fractions relies on cold. However, lately, ambient temperature preservation has been evaluated by several independent institutions that see economic and logistic advantages in getting rid of the cold chain. Here we validated a novel procedure for ambient temperature preservation of DNA in white blood cell and buffy coat lysates based on the confinement of the desiccated biospecimens under anoxic and anhydrous atmosphere in original hermetic minicapsules. For this validation we stored encapsulated samples either at ambient temperature or at several elevated temperatures to accelerate aging. We found that DNA extracted from stored samples was of good quality with a yield of extraction as expected. Degradation rates were estimated from the average fragment size of denatured DNA run on agarose gels and from qPCR reactions. At ambient temperature, these rates were too low to be measured but the degradation rate dependence on temperature followed Arrhenius’ law, making it possible to extrapolate degradation rates at 25°C. According to these values, the DNA stored in the encapsulated blood products would remain larger than 20 kb after one century at ambient temperature. At last, qPCR experiments demonstrated the compatibility of extracted DNA with routine DNA downstream analyses. Altogether, these results showed that this novel storage method provides an adequate environment for ambient temperature long term storage of high molecular weight DNA in dehydrated lysates of white blood cells and buffy coats.

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“…The first is anhydrobiosis, the dehydration process used by some organisms to Physically protecting nucleic acids from the environment, through encapsulation or adsorption onto a solid support, is the third of the stabilization principles and has emerged for the delivery of gene therapeutics. Materials, such as liposomes, metal particles, mesoporous silica nanoparticles, polymers, potato starch, silk fibron and surfactants, have been developed with these applications in mind [35][36][37][38][39][40][41][42][43][44][45][46][47]. In a previous study, we demonstrated the potential of RNA adsorption onto mesoporous silica nanoparticles (MSNs) with and without additional stabilizing reagents.…”

Section: Introductionmentioning

confidence: 99%

Adsorption and Elution of Nucleic Acids: Mesoporous Materials and Methods

Lin¹,

Johnson²,

Melde³

et al. 2017

oajost

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Physical protection of nucleic acids from the environment for improved stability through encapsulation or adsorption has been reported using various materials, including liposomes, metal particles, mesoporous silica nanoparticles, and polymers. In an extension of that approach, our previous study demonstrated the potential of mesoporous silica nanoparticles with and without covalently attached stabilizing reagents, such as sugar and bovine serum albumin, for improving the stability of RNA. The goal of the current study is to evaluate the potential of silicate sorbents bearing chemical functionalities for stabilization of nucleic acid targets. Materials offering charged groups, metal chelating sites, and -bonding sites are considered. Adsorption and elution of RNA, DNA, and single stranded DNA (ssDNA) are evaluated as is subsequent elution of the bound target. A sorbent functionalized with primary amine groups showed promising results for RNA and ssDNA stabilization. The impact of the sorbents on long term viability the targets is also evaluated. Storage of adsorbed targets at room temperature and 37 ∘ C over a period of 200 d indicates the potential for stabilization of RNA and ssDNA using several of the functionalities. None of the sorbents improved the stability of DNA either under room temperature or 37 ∘ C storage.

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A novel method for room temperature distribution and conservation of RNA and DNA reference materials for guaranteeing performance of molecular diagnostics in onco-hematology: A GBMHM study

Cited by 5 publications

References 16 publications

Ensuring the Safety and Security of Frozen Lung Cancer Tissue Collections through the Encapsulation of Dried DNA

Ensuring the Safety and Security of Frozen Lung Cancer Tissue Collections through the Encapsulation of Dried DNA

High DNA stability in white blood cells and buffy coat lysates stored at ambient temperature under anoxic and anhydrous atmosphere

Adsorption and Elution of Nucleic Acids: Mesoporous Materials and Methods

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