1.Development of Indicators to Assess the Stability of Remnant Blood Samples Stored in a Biobank: Experience at One Institution.
Sae Hwan KIM ; Young Eun KANG ; Young Jun HONG ; Yoon Hwan CHANG ; Seok Il HONG ; Ae Chin OH ; Jin Kyung LEE
The Korean Journal of Laboratory Medicine 2010;30(6):718-725
BACKGROUND: One of the major concerns with biobanking is the absence of standard operating procedures to eliminate pre-analytical variation arising from sample collection, preparation, and storage. Currently, there is a lack of tools to carry out quality control procedures for stored blood samples. The aim of this study is to assess the quality of stored blood samples in our biobank and to suggest appropriate indicators for their quality control. METHODS: The stored blood samples that we tested have been registered into our biobank since 2003. These were transferred to our biobank after carrying out routine requested tests, because the samples would have otherwise been discarded. For the purpose of quality control, we analyzed the concentrations and the integrity of DNA and RNA extracted from the stored samples and tested the levels of several serum proteins; the results were compared with the corresponding pre-storage levels. RESULTS: A total of 19 samples were stored from 2006 to 2009. Of the 22 samples stored between 2003 and 2005, 50% showed complete DNA integrity. However, sufficient RNA integrity was noted in only 1 sample stored as recently as 2009. High blood urea nitrogen levels were also noted in the stored sera, but the increase did not correlate to the duration of storage. CONCLUSIONS: The amount and integrity of nucleic acids extracted from stored blood samples are potential indicators that can be used for quality control. A guideline for the quality assessment of stored blood samples in a biobank is urgently needed.
Blood Banks/*standards
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Blood Proteins/chemistry/standards
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Blood Urea Nitrogen
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DNA/*analysis/chemistry/standards
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Laboratory Techniques and Procedures
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Quality Control
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RNA/*analysis/chemistry/standards
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Specimen Handling/methods
2.Comparison of Two New Plastic Tubes (Sekisui INSEPACK and Green Cross Green Vac-Tube) with BD Vacutainer Tubes for 49 Analytes.
The Korean Journal of Laboratory Medicine 2007;27(1):69-75
BACKGROUND: Becton Dickinson (BD) Vacutainer tubes are the most widely used vacuum system for collection of blood in clinical laboratories. We compared the performance of two new tubes, Sekisui INSEPACK tube and Green Cross Green Vac-Tube, with the existing BD Vacutainer tubes for 49 common analytes. METHODS: A total of 20 apparently healthy volunteers were recruited for this study. For rountine chemistry and thyroid function tests, we compared the results of two new vacutainer tubes and BD Vacutainer tubes with those of BD glass tubes at t =0 hr by student paired t test. Hematology and coagulation test results of the two new vacutainer tubes were compared with those of BD Vacutainer tubes at t =0 hr. To study the stability of each analyte, results at t =24 +/- 2 hr, t =72 +/- 2 hr, and t =168 +/- 2 hr were compared with those at t =0 hr for each tube. RESULTS: Although paired t test analysis revealed statistically significant differences between two tested tubes and existing BD Vacutainer tube in several tests (total protein, total bilirubin, alkaline phosphatase, glucose, uric acid, calcium, inorganic phosphorus, direct bilirubin, triglyceride, HDL-cholesterol, LDL-cholesterol, lactate dehydrogenase, triiodothyronine, and thyroxine), these differences were not considered clinically significant. Stability of two new vacuum tubes for each analyte was similar to that of the BD Vacutainer tube. CONCLUSIONS: Sekisui INSEPACK tube and Green Cross Green-Vac Tube showed a satisfactory analytical performance compared with existing BD Vacutainer tubes. We conclude that these two new plastic vacutainer tubes are acceptable for the commonly ordered laboratory tests.
Blood Cells/chemistry
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Blood Proteins/analysis
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Blood Specimen Collection/*instrumentation
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Equipment and Supplies/standards
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Female
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Humans
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Male
3.Establishment and characterization of an infectious cDNA clone of a classical swine fever virus LOM strain.
Gil Soon PARK ; Seong In LIM ; Seung Ho HONG ; Jae Young SONG
Journal of Veterinary Science 2012;13(1):81-91
Classical swine fever virus (CSFV) causes a highly contagious disease among swine that has an important economic impact worldwide. CSFV strain LOM is an attenuated virus of low virulent strain of Miyagi isolated from Japan in 1956. Eight DNA fragments representing the genome of the CSFV strain LOM were obtained by RT-PCR. These were used to determine the complete nucleotide sequence and construct a full-length cDNA clone which was called Flc-LOM. Sequence analysis of the recombinant clone (Flc-LOM) revealed the presence of eight mutations, resulting in two amino acid substitutions, when compared to the parental sequence. RNA transcripts of both LOM and Flc-LOM were directly infectious in PK-15 cells. The rescued Flc-LOM virus grew more slowly than the parental virus, LOM, in the cells. Intramuscular immunization with Flc-LOM was safe and highly immunogenic in pigs; no clinical signs or virus transmission to sentinel animals were observed after 35 days. CSFV-specific neutralizing antibodies were detected 14 days post-infection. After challenge with the virulent CSFV strain SW03, pigs immunized with Flc-LOM were shown to be fully protected. Thus, our newly established infectious clone of CSFV, Flc-LOM, could serve as a vaccine candidate.
Animals
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Antibodies, Viral/blood
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Base Sequence
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Cell Line
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Classical Swine Fever/immunology/*virology
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Classical swine fever virus/*genetics/immunology/pathogenicity
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Cloning, Molecular
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DNA, Complementary/genetics/immunology
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Immunization/methods/standards/veterinary
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Molecular Sequence Data
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Neutralization Tests/veterinary
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RNA, Viral/chemistry/genetics
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Recombinant Proteins/immunology
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Reverse Transcriptase Polymerase Chain Reaction/veterinary
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Sequence Analysis, DNA
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Specific Pathogen-Free Organisms
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Swine
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Virulence