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 ; Blood Proteins/chemistry/standards ; Blood Urea Nitrogen ; DNA/*analysis/chemistry/standards ; Laboratory Techniques and Procedures ; Quality Control ; RNA/*analysis/chemistry/standards ; Specimen Handling/methods

Viral load testing of human immunodeficiency virus (HIV) is an essential tool for initiating and monitoring the antiretroviral therapy for HIV patients. To this end, several methods including polymerase chain reaction (PCR), branched DNA (bDNA), nucleic acid sequence based amplification assay (NASBA) and internally controlled virion PCR (ICV PCR) have become available. Of these methods, the standard reverse transcription-PCR (RT-PCR) assay has been widely used in Korea. However, no comparison study has been performed among the various detection methods currently used in Korean patients. We evaluated the correlation and agreement between the PCR and the branched DNA (bDNA) assay for measurement of HIV RNA in Korean patients. Eighty randomly selected samples from HIV-1-seropositive patients visiting Yonsei Medical Center Severance Hospital were studied. We found that these assays show good agreement, have a reliable correlation (r = 0.92, mean difference in log10 copies/ mL +/- 2 standard deviation = 0.098 +/- 0.805) and produce values whose relationship is given by the following equation: log10v3 bDNA = -0.3405 + 1.0601 x log10RT-PCR. Thus, we conclude that these two methods may allow direct comparison of the results obtained from different assay systems.
Comparative Study ; DNA/chemistry* ; DNA/analysis* ; Genetic Techniques/standards* ; HIV-1/genetics* ; Human ; Korea ; RNA, Viral/blood* ; Reverse Transcriptase Polymerase Chain Reaction/standards*

Age Factors ; Alanine Transaminase ; blood ; Blood Glucose ; analysis ; Body Mass Index ; Chemistry, Clinical ; standards ; Female ; Hepatitis B, Chronic ; blood ; pathology ; Hepatitis C, Chronic ; blood ; pathology ; Humans ; Lipids ; blood ; Liver Diseases ; blood ; pathology ; Male ; RNA, Viral ; blood ; Reference Values ; Retrospective Studies ; Sex Factors

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 ; Antibodies, Viral/blood ; Base Sequence ; Cell Line ; Classical Swine Fever/immunology/*virology ; Classical swine fever virus/*genetics/immunology/pathogenicity ; Cloning, Molecular ; DNA, Complementary/genetics/immunology ; Immunization/methods/standards/veterinary ; Molecular Sequence Data ; Neutralization Tests/veterinary ; RNA, Viral/chemistry/genetics ; Recombinant Proteins/immunology ; Reverse Transcriptase Polymerase Chain Reaction/veterinary ; Sequence Analysis, DNA ; Specific Pathogen-Free Organisms ; Swine ; Virulence