Influence of the storage temperature on the nucleic acid quality in liquid-based cytology samples prepared using fixed in CytoRich Red

2021 ◽  
Vol 60 (4) ◽  
pp. 212-218
Author(s):  
Hideyuki ABE ◽  
Akihiko KAWAHARA ◽  
Eiji SADASHIMA ◽  
Kazuya MURATA ◽  
Yorihiko TAKASE ◽  
...  
Keyword(s):  
Nucleic Acid ◽  
Storage Temperature ◽  
Liquid Based Cytology

scholarly journals Rapid Bedside Inactivation of Ebola Virus for Safe Nucleic Acid Tests

2016 ◽  
Vol 54 (10) ◽  
pp. 2521-2529 ◽  
Author(s):  
Maiken Worsøe Rosenstierne ◽  
Helen Karlberg ◽  
Karoline Bragstad ◽  
Gunnel Lindegren ◽  
Malin Lundahl Stoltz ◽  
...  
Keyword(s):  
Nucleic Acid ◽  
Ebola Virus ◽  
Storage Temperature ◽  
Nucleic Acid Detection ◽  
Blood Collection ◽  
Acid Extraction ◽  
Simple Method ◽  
Virus Rna ◽  
Vacuum Tubes ◽  
Nucleic Acid Tests

Rapid bedside inactivation of Ebola virus would be a solution for the safety of medical and technical staff, risk containment, sample transport, and high-throughput or rapid diagnostic testing during an outbreak. We show that the commercially available Magna Pure lysis/binding buffer used for nucleic acid extraction inactivates Ebola virus. A rapid bedside inactivation method for nucleic acid tests is obtained by simply adding Magna Pure lysis/binding buffer directly into vacuum blood collection EDTA tubes using a thin needle and syringe prior to sampling. The ready-to-use inactivation vacuum tubes are stable for more than 4 months, and Ebola virus RNA is preserved in the Magna Pure lysis/binding buffer for at least 5 weeks independent of the storage temperature. We also show that Ebola virus RNA can be manually extracted from Magna Pure lysis/binding buffer-inactivated samples using the QIAamp viral RNA minikit. We present an easy and convenient method for bedside inactivation using available blood collection vacuum tubes and reagents. We propose to use this simple method for fast, safe, and easy bedside inactivation of Ebola virus for safe transport and routine nucleic acid detection.

scholarly journals Influence of Storage Conditions on SARS-CoV-2 Nucleic Acid Detection in Throat Swabs

2020 ◽  
Vol 222 (2) ◽  
pp. 203-205 ◽  
Author(s):  
Lin Li ◽  
Xiao Li ◽  
Zhendong Guo ◽  
Zhongyi Wang ◽  
Ke Zhang ◽  
...  
Keyword(s):  
Nucleic Acid ◽  
Storage Time ◽  
Storage Temperature ◽  
Storage Conditions ◽  
Nucleic Acid Detection ◽  
Nucleic Acid Testing ◽  
Threshold Values ◽  
Sample Storage ◽  
Cycle Threshold ◽  
And Storage

Abstract The detection of SARS-CoV-2 infection is the premise of quarantine. In many countries or areas, samples need to be shipped or inactivated before SARS-CoV-2 testing. In this study, we checked the influence of sample storage conditions on SARS-CoV-2 nucleic acid testing results, including sample inactivation time, storage temperature, and storage time. All of these conditions caused an increase in the cycle threshold values of the nucleic acid tests and led to the misclassification of at least 10.2% of positive cases as negative or suspected. The results highlight the importance of immediate testing of samples for SARS-CoV-2 nucleic acid detection.

Efficacy of the Antigenicity-Retaining Ability of Fixative Solutions for Liquid-Based Cytology: Immunocytochemistry of Long-Term Storage

2021 ◽  
pp. 1-12
Author(s):  
Hiaki Sato ◽  
Yoshiaki Norimatsu ◽  
Satoshi Irino ◽  
Takeshi Nishikawa
Keyword(s):  
Cell Membrane ◽  
Room Temperature ◽  
Storage Temperature ◽  
Storage Period ◽  
Storage Conditions ◽  
Immunocytochemical Staining ◽  
Long Term Storage ◽  
Liquid Based Cytology ◽  
Term Storage

<b><i>Introduction/Objective:</i></b> Liquid-based cytology (LBC) is advantageous as multiple stained specimens can be prepared and used for additional assays such as immunocytochemical and molecular-pathological investigations. Two types of preservative-fixative solutions (fixatives) are used for nongynecologic specimens used in the BD SurePath-LBC (SP-LBC) method, and their components vary. However, few studies have evaluated the differences in antigen-retaining ability between these fixatives. Therefore, we investigated and compared the antigen-retaining ability of the fixatives in immunocytochemical staining (ICC) under long-term storage conditions. <b><i>Materials and Methods:</i></b> Sediments of cultured RAJI cells (derived from Burkitt’s lymphoma) were added to each fixative (red and blue) and stored at room temperature for a specified period (1 h; 1 week; and 1, 3, and 6 months). The specimens were then prepared using the SP-LBC method and subjected to ICC. Positivity rate was calculated using the specimens fixed at room temperature for 1 h as a control. Antibodies against Ki67 expressed in the nucleus and against CD20 and leukocyte common antigen (LCA) expressed on the cell membrane were used. <b><i>Results:</i></b> For CD20 and LCA, the positivity rate increased with time in the red fixative compared with that in the control. In the blue fixative, the positivity rate was highest at 1 h and was maintained at a high level throughout the storage period. In contrast, the Ki67 positivity rate was highest at 1 h in both red and blue fixatives and markedly decreased with time. Therefore, although refrigerated (8°C) storage was used, no improvement was noted. <b><i>Conclusions:</i></b> Long-term storage is possible for cell membrane antigens at room temperature; however, it is unsuitable for intranuclear antigens. Therefore, we conclude that suitable fixative type and storage temperature differ based on antigen location. Further investigation is warranted.

scholarly journals Effects of Storage Temperature and Media/Buffer for SARS-CoV-2 Nucleic Acid Detection

2020 ◽  
Author(s):  
Namhee Kim ◽  
Ahrin Kwon ◽  
Eun Youn Roh ◽  
Jong Hyun Yoon ◽  
Mi Seon Han ◽  
...  
Keyword(s):  
Nucleic Acid ◽  
Storage Temperature ◽  
Medical Center ◽  
Quantitative Polymerase Chain Reaction ◽  
Storage Period ◽  
Storage Conditions ◽  
Nucleic Acid Detection ◽  
Lower Probability ◽  
Sample Collection ◽  
Number Of Patients

Abstract Objectives The increase in the number of patients with coronavirus disease 2019 (COVID-19) has delayed real-time reverse transcription–quantitative polymerase chain reaction (RT-qPCR), requiring proper shipping and storage conditions, especially in hot weather. This study aims to assess how some conditions, such as storage period, temperature, media or buffer, and sample types, affect the results of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) RT-qPCR. Methods SARS-CoV-2–positive specimens were collected from Boramae Medical Center for 2 months (from May to June 2020) and stored in different media or buffers at different temperatures. Results As a result of examining confirmed patient samples, RT-qPCR results were not significantly affected by 2°C to 8°C storage until after 7 days. When stored at 20°C to 22°C or above 35°C, the results were affected negatively even after 1 day. Higher storage temperatures resulted in a lower probability of detecting viral nucleic acids because of degradation. Samples stored in pH-controlled media or buffer were more stable than those stored in nonbuffer states. Conclusions These results emphasize the importance of storage temperature and media or buffer and performing RT-qPCR for SARS-CoV-2 nucleic acid detection as soon as possible after sample collection.

Nucleic acid quality in sodium alginate cell blocks prepared from liquid-based cytology specimens

2021 ◽  
Vol 60 (2) ◽  
pp. 102-109
Author(s):  
Hideyuki ABE ◽  
Akihiko KAWAHARA ◽  
Eiji SADASHIMA ◽  
Ryota TANAKA ◽  
Kazuya MURATA ◽  
...  
Keyword(s):  
Nucleic Acid ◽  
Sodium Alginate ◽  
Liquid Based Cytology

Introduction

1978 ◽  
Vol 36 (3) ◽  
pp. 543-544
Author(s):  
W. Bernard
Keyword(s):  
Molecular Weight ◽  
Electron Microscope ◽  
Nucleic Acid ◽  
Gene Mapping ◽  
Molecular Genetics ◽  
Cell Biology ◽  
Gene Activity ◽  
Close Connection ◽  
Basic Information ◽  
Simple Systems

In comparison to many other fields of ultrastructural research in Cell Biology, the successful exploration of genes and gene activity with the electron microscope in higher organisms is a late conquest. Nucleic acid molecules of Prokaryotes could be successfully visualized already since the early sixties, thanks to the Kleinschmidt spreading technique - and much basic information was obtained concerning the shape, length, molecular weight of viral, mitochondrial and chloroplast nucleic acid. Later, additonal methods revealed denaturation profiles, distinction between single and double strandedness and the use of heteroduplexes-led to gene mapping of relatively simple systems carried out in close connection with other methods of molecular genetics.

Infection of Escherichia coli with bacteriophages

1969 ◽  
Vol 27 ◽  
pp. 370-371
Author(s):  
Manfred E. Bayer
Keyword(s):  
Escherichia Coli ◽  
Nucleic Acid ◽  
Cell Surface ◽  
Virus Type ◽  
Infection Process ◽  
Adsorption Site ◽  
The Other ◽  
Specific Receptors ◽  
Bacterial Receptors

The first step in the infection of a bacterium by a virus consists of a collision between cell and bacteriophage. The presence of virus-specific receptors on the cell surface will trigger a number of events leading eventually to release of the phage nucleic acid. The execution of the various "steps" in the infection process varies from one virus-type to the other, depending on the anatomy of the virus. Small viruses like ØX 174 and MS2 adsorb directly with their capsid to the bacterial receptors, while other phages possess attachment organelles of varying complexity. In bacteriophages T3 (Fig. 1) and T7 the small conical processes of their heads point toward the adsorption site; a welldefined baseplate is attached to the head of P22; heads without baseplates are not infective.

Nucleic Acid Molecules Prepared by Monolayer Techniques

1972 ◽  
Vol 30 ◽  
pp. 178-179
Author(s):  
Dimitrij Lang
Keyword(s):  
Electron Microscopy ◽  
Standard Deviation ◽  
Nucleic Acid ◽  
Cytochrome C ◽  
Nucleic Acids ◽  
Contour Length ◽  
Sample Standard Deviation ◽  
Molecular Weights ◽  
Length Measurements ◽  
Protein Monolayer

The success of the protein monolayer technique for electron microscopy of individual DNA molecules is based on the prevention of aggregation and orientation of the molecules during drying on specimen grids. DNA adsorbs first to a surface-denatured, insoluble cytochrome c monolayer which is then transferred to grids, without major distortion, by touching. Fig. 1 shows three basic procedures which, modified or not, permit the study of various important properties of nucleic acids, either in concert with other methods or exclusively:1) Molecular weights relative to DNA standards as well as number distributions of molecular weights can be obtained from contour length measurements with a sample standard deviation between 1 and 4%.

Snapshot blotting: The transfer of nucleic acids and nucleoprotein complexes from electrophoresis gels to EM grids

1992 ◽  
Vol 50 (1) ◽  
pp. 762-763
Author(s):  
Stephen D. Jett
Keyword(s):  
Nucleic Acid ◽  
Nucleic Acids ◽  
Nucleic Acid Binding ◽  
Mobility Shift ◽  
Carbon Coated ◽  
Nucleoprotein Complexes ◽  
Mobility Shift Assay ◽  
Protein Nucleic Acid ◽  
Gel Mobility Shift ◽  
Nucleic Acid Interactions

The electrophoresis gel mobility shift assay is a popular method for the study of protein-nucleic acid interactions. The binding of proteins to DNA is characterized by a reduction in the electrophoretic mobility of the nucleic acid. Binding affinity, stoichiometry, and kinetics can be obtained from such assays; however, it is often desirable to image the various species in the gel bands using TEM. Present methods for isolation of nucleoproteins from gel bands are inefficient and often destroy the native structure of the complexes. We have developed a technique, called “snapshot blotting,” by which nucleic acids and nucleoprotein complexes in electrophoresis gels can be electrophoretically transferred directly onto carbon-coated grids for TEM imaging.

High-resolution nucleic acid sequence mapping via in situ hybridization at the Electron Microscope level

1995 ◽  
Vol 53 ◽  
pp. 752-753
Author(s):  
B.A. Hamkalo ◽  
S. Narayanswami ◽  
A.P. Kausch
Keyword(s):  
Nucleic Acid ◽  
Interphase Nucleus ◽  
Genome Mapping ◽  
Precise Location ◽  
Electron Microscope Level ◽  
Rna Sequences ◽  
Fundamental Interest ◽  
Whole Cells ◽  
Dna And Rna

The availability of nonradioactive methods to label nucleic acids an the resultant rapid and greater sensitivity of detection has catapulted the technique of in situ hybridization to become the method of choice to locate of specific DNA and RNA sequences on chromosomes and in whole cells in cytological preparations in many areas of biology. It is being applied to problems of fundamental interest to basic cell and molecular biologists such as the organization of the interphase nucleus in the context of putative functional domains; it is making major contributions to genome mapping efforts; and it is being applied to the analysis of clinical specimens. Although fluorescence detection of nucleic acid hybrids is routinely used, certain questions require greater resolution. For example, very closely linked sequences may not be separable using fluorescence; the precise location of sequences with respect to chromosome structures may be below the resolution of light microscopy(LM); and the relative positions of sequences on very small chromosomes may not be feasible.

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