No abstract available.
Paramyxoviridae Infections* ; Polymerase Chain Reaction* ; Reverse Transcription*

BACKGROUND: Kruppel-like factor 4 (KLF4) is a transcription factor that regulates a diverse array of cellular processes, including development, differentiation, proliferation, and apoptosis. Although its function in keratinocytes has been widely studied, its exact role in psoriasis has not been elucidated. OBJECTIVE: We designed this study to investigate epidermal expression levels of KLF4 and the change in KLF4 expression after treatment in patients with psoriasis. METHODS: We compared the expression levels of KLF4 in the basal, suprabasal, and superficial epidermal layers, in psoriatic lesional, non-lesional, and normal skin, using an immunoreactivity intensity distribution index (IRIDI). In addition, we measured the change in KLF4 expression on the basis of the IRIDI and by reverse transcription polymerase chain reaction (RT-PCR) analysis after treatment. RESULTS: The combined IRIDI scores in psoriatic lesional skin were significantly higher than the scores in both non-lesional and normal skin. The psoriatic epidermis, particularly the suprabasal layer, showed a significantly increased IRIDI score compared to that of non-lesional and normal skin, which was significantly decreased after treatment. RT-PCR analysis exhibited a slight increase in KLF4 mRNA expression level after treatment; however, this increase was not significant. CONCLUSION: These data indicate that KLF4 could regulate epidermal proliferation and differentiation. Moreover, we believe that KLF4 may play an important role in the physiological reaction to counteract abnormal differentiation and proliferation of keratinocytes.
Apoptosis ; Epidermis ; Humans ; Keratinocytes ; Polymerase Chain Reaction ; Psoriasis* ; Reverse Transcription ; RNA, Messenger ; Skin ; Transcription Factors

BACKGROUND: Little Is known about the compared efficiency of different third generation enzyme-linked immunosorbent assays (ELISA) fort the detection of anti-HCV. We examine the relative sensitivity and specificity of three third-generation anti-HCV assays, and results of discrepant samples among the anti-HCV ELISA are compared with data of a third-generation recombinant immunoblot assay and reverse transcription polymerase chain reaction (RT-PCR) . METHODS:A total of 167 samples (61 positive and 106 negative), screened by a second-generation IMx(R) anti-HCV assay (Abbott 2.0; Abbott Laboratories, USA), weve tested with Innotest HCV 3.0(R) (Green Cross, Korea), LG HCD 3.0(R) (LG, Korea) and DONG-A HCV 3.0(R) (Dong-4, Korea). The discrepant specimens among the 4 anti-HCV ELISA were tested by LG HCD Confirm(R) (LG, Korea) and RT-PCR. RESULTS: The concordance rates of all 4 ansi-HCV ELISA were 80.2% (134/167) and 92.2% (154/167), respectively. The 28 and 31 of 33 specimens showing discrepancy among 4 anti-HCV ELISA were tested with LG HCD Confirm and RT-PCR, respectively. Serum HCV RNA was positive in 2 of 2 reactive and in 6 of 26 nonreactive on LG HCD Confirm. The sensitivity, specificity, positive predictive value, negative predictive value and concordance rate of 4 anti-HCV ELISA were 97.7%, 85.2%, 70.0%, 99.0% and 88.5% (Abbott 2.0) ; 81.4%, 96.7%, 89.7%, 93.7% and 92 7% (Innotest 3.0), 81.4%, 98.4%, 94.6%, 93.8% and 93.9% (LG 3.0), 86.0%, 95.7%, 88.1%, 95.1% and 93.3% (DONG-A 3.0), respectively. CONCLUSIONS: These data indicate that the sensitivity and specificity of 3 third-generation anti-HCV ELISA are comparable, and that these reagents demonstrate improved specificity compared to the second-generation ELISA.
Enzyme-Linked Immunosorbent Assay* ; Indicators and Reagents ; Polymerase Chain Reaction ; Reverse Transcription ; RNA ; Sensitivity and Specificity

BACKGROUND: The detection of total anti-hepatitis A virus (anti-HAV) immunoglobulin (Ig) and IgM is important for diagnosing acute hepatitis A. Our laboratory introduced new commercial automated chemiluminescence immunoassays (CLIAs) for use in addition to pre-existing automated CLIA. We evaluated the rate of agreement in the detection of total anti-HAV Ig and IgM in serum samples between two automated CLIAs. METHODS: We analyzed 181 samples those were submitted for testing at Kangbuk Samsung Medical Center. We analyzed the rate of agreement between the ADVIA Centaur XP (Siemens, Germany) and the MODULAR ANALYTICS E170 (Roche, Switzerland) analyzers. We performed reverse transcription (RT)-PCR when there was a discrepancy between the results from the two analyzers. RESULTS: The agreement rates between the ADVIA Centaur XP and the MODULAR ANALYTICS E170 for total anti-HAV Ig and IgM were 97.2% and 98.9%, respectively. Discrepant results were obtained in seven cases; all were found to be HAV-negative based on RT-PCR analysis. CONCLUSIONS: The total anti-HAV Ig and IgM results obtained using the two automated analyzers were comparable. However, in cases of equivocal results tested by the ADVIA Centaur XP for anti-HAV IgM, retesting and follow-up testing of samples are recommended.
Hepatitis A ; Hepatitis A Antibodies ; Hepatitis A virus ; Immunoassay ; Immunoglobulin M ; Immunoglobulins ; Luminescence ; Reverse Transcription ; Viruses

PURPOSE: Since the discovery of estrogen receptor-beta(ER-beta, five C-terminal variants of ER-beta were identified. We designed this study to investigate the pattern and clinical implications of ER-betaand its splicing variants expression in normal and malignant mammary tissues. METHODS: Using reverse transcription polymerase chain reaction (RT-PCR), we examined the expression levels of ER-alpha and ER-betaand its five splicing variants (beta1, beta2, beta3, beta4, beta5) in 50 paired normal and cancer tissues. We measured the densities of RT-PCR products using Tina version 2.10 (Raytest, Germany). Firstly, the incidence and intensity of ER-alpha and ER-beta and its five splicing variants were compared. Then the expression of ER-betamRNA splicing variants was also analyzed with regard to the ER-alphaprotein expression measured by immuno-histochemical staining and the menopausal status of the patients. Chi-square test and paired samples t-test were used for statistical analysis. Differences were considered to be significant with a p-value of less than 0.05. RESULTS: The expression of ER-betamRNA variants in normal breast and cancer tissues were as follows: ER-beta2 (100%/100%), ER-beta4 (76%/74%), ER-beta5 (32%/58%), and ER-beta1 (14%/16%). ER-beta3 was not detected at all. In terms of intensity, we observed a significant decrease of ER-beta2 (P<0.001) and an increase of ER-beta5 (P=0.004) in the mRNA expression levels among breast cancers compared to the corresponding normal breast tissues. Compared to the corresponding normal tissues, a significant decrease of ER-beta2 in cancer tissues was observed in patients with ER-alpha-positive (P<0.001), with age over 50 (P=0.01), and under 50 (P=0.04) as well, but not in patients with ER-alpha-negative (P=0.48). ER-beta4 also significantly decreased in patients with ER-alpha-positive (P=0.004) and with age over 50 (P=0.07). ER-beta5 showed a significant increment only in patient aged over 50 (P=0.04). CONCLUSION: ER-alpha mRNA expression significantly increases but ER-beta mRNA expression decreases in the cancer tissues compared to the corresponding normal tissues. Among ER-beta variant forms, ER-beta2 is predominant in both normal and malignant mammary tissues and ER-beta4, ER-beta5, and ER-beta1 in descending order but ER-beta3 does not express in mammary tissues. The decrease of ER-beta2 and ER-beta4 expression is prominent in cancer tissue especially in ER-alpha-positive cancers, which suggests that ER-beta2 and ER-beta4 may possess a regulatory function in mammary carcinogenesis. Further investigations to verify the roles of ER-beta variants are mandatory.
Breast ; Breast Neoplasms ; Carcinogenesis ; Estrogens* ; Humans ; Incidence ; Polymerase Chain Reaction ; Receptors, Estrogen ; Reverse Transcription ; RNA, Messenger

BACKGROUND: The detection of antibody to hepatitis C virus (anti-HCV) is the most useful method to investigate past or current HCV infections. We performed a clinical evaluation of ADVIA Centaur HCV assay, a new third generation assay for the qualitative detection of IgG antibody. METHODS: Included in the study were a total of 323 samples (108 positive and 215 negative), for which HCV reverse transcription polymerase chain reaction (RT-PCR) was requested. ADVIA Centaur HCV assay (Bayer Healthcare LLC, Diagnostics Division, Tarrytown, NY, USA) was compared with a currently available and widely used AxSYM HCV version 3.0 assay (Abbott Laboratories, Abbott Park, IL, USA). Samples with discrepant results were retested with each assay, and further tested with a recombinant immunoblot assay (RIBA, LG HCD Confirm, LG Chemical Co., Seoul, Korea). Reproducibility of Centaur assay was evaluated in five groups (two samples in each group) with different index values. RESULTS: The overall concordance rate was 91.6% (296/323) between Centaur and AxSYM assays. It was 100% (108/108) in RT-PCR positive samples and 87.4% (188/215) in RT-PCR negative samples. Discrepant samples (8.4%, 27/323) were all RT-PCR negative, and all except two were Centaur negative and AxSYM positive. In discrepant samples, RIBA showed negative results except for two samples with indeterminate results. The sensitivity and specificity of Cenyaur assay were 98.1% and 65.6%, and the respective figures for AxSYM assay were 98.1% and 54.9%. Reproducibility of Centaur assay was satisfactory. CONCLUSIONS: The overall concordance between Centaur and AxSYM assays was satisfactory. Sensitivity and specificity of Centaur assay were equivalent to or better than those of AxSYM assay. ADVIA Centaur HCV assay seems to be a reliable and useful method for the detection of anti-HCV in clinical laboratories.
Delivery of Health Care ; Hepacivirus* ; Immunoglobulin G ; Polymerase Chain Reaction ; Reverse Transcription ; Sensitivity and Specificity ; Seoul

A retron is a bacterial retroelement that encodes an RNA gene and a reverse transcriptase (RT). The former, once transcribed, works as a template primer for reverse transcription by the latter. The resulting DNA is covalently linked to the upstream part of the RNA; this chimera is called multicopy single-stranded DNA (msDNA), which is extrachromosomal DNA found in many bacterial species. Based on the conserved features in the eight known msDNA sequences, we developed a detection method and applied it to scan National Center for Biotechnology Information (NCBI) RefSeq bacterial genome sequences. Among 16,844 bacterial sequences possessing a retron-type RT domain, we identified 48 unique types of msDNA. Currently, the biological role of msDNA is not well understood. Our work will be a useful tool in studying the distribution, evolution, and physiological role of msDNA.
Biotechnology ; Chimera ; DNA ; DNA, Single-Stranded* ; Genome, Bacterial* ; Retroelements ; Reverse Transcription ; RNA ; RNA-Directed DNA Polymerase

The incidence of severe fever with thrombocytopenia syndrome (SFTS) has increased in Korea since a first report in 2013. We investigated whether SFTS existed before 2013 using real-time reverse transcription polymerase chain reaction and stored blood samples from febrile patients with thrombocytopenia. Four cases of SFTS were identified, with the earliest occurring in 2008.
Fever* ; Humans ; Incidence ; Korea* ; Lymphohistiocytosis, Hemophagocytic* ; Phlebovirus ; Polymerase Chain Reaction ; Retrospective Studies* ; Reverse Transcription ; Thrombocytopenia*

BACKGROUND/AIMS: Ghrelin levels are known to increase in patients with ulcerative colitis (UC), but serum obestatin levels in UC patients are not well elucidated. The aim of this study was to examine the relationship between serum ghrelin and obestatin levels and disease activity in UC patients. METHODS: The serum ghrelin and obestatin levels were measured in 21 UC patients (12 with active disease and 9 in remission) using enzyme-linked immunosorbent assay. The relationship between the circulating levels of these 2 hormones and disease activity was analyzed. The colonic mucosal mRNA expression of ghrelin and obestatin was measured by quantitative reverse transcription polymerase chain reaction. RESULTS: The mean serum ghrelin values were significantly higher in patients with active disease than in patients with remission (1370.6+/-404.3 vs. 783.5+/-235.3 pg/mL, P=0.001). Colonic mucosal mRNA expression of ghrelin was also significantly higher in patients with active disease than in patients in remission (0.805+/-0.214 vs. 0.481+/-0.356, P=0.018). However, the mean serum obestatin levels and colonic mucosal mRNA expression of obestatin were not significantly different between both groups. The circulating obestatin/ghrelin ratio was significantly lower in patients with active UC than in patients in remission (0.32+/-0.08 vs. 0.58+/-0.20, P=0.001). CONCLUSIONS: The serum ghrelin levels and the obestatin/ghrelin ratio were related to the activity of UC, but serum obestatin was not related to activity of UC. The ghrelin levels and the obestatin/ghrelin ratio could serve as activity markers in patients with UC.
Colitis, Ulcerative* ; Colon ; Enzyme-Linked Immunosorbent Assay ; Ghrelin* ; Humans ; Polymerase Chain Reaction ; Reverse Transcription ; RNA, Messenger

Murine norovirus (MNV) is a non-enveloped virus with a positive-sense RNA genome and causes lethal infection in mice. MNV has been used as a model virus for human norovirus (NV) whose in vitro cell culture system has not been available to date since MNV and NV are genetically related. In this study, the genome replication of MNV was investigated using strand-specific RT-PCR in RAW264.7 cells. Reverse transcription (RT) using a sense primer followed by PCR showed that negative-sense RNAs were first detected in RAW264.7 cells between 6 and 9 [3 and 6] hours post infection (h.p.i.). However, these negative-sense RNAs were not detected when cells were treated with a translation inhibitor cycloheximide. Then, RT with an antisense primer followed by PCR was performed to detect positive-sense RNAs. RT-PCR results revealed that the amount of positive-sense RNAs began to increase from 9 [6] h.p.i., indicating the accumulation of the newly synthesized (+)RNA genome. Furthermore, cycloheximide abrogated the increase of newly made RNAs during MNV infection. In conclusion, strand-specific RT-PCR using a sense or antisense primer, in combination with cycloheximide treatment, enabled us to detect positive-sense and negative-sense RNAs selectively and provided a useful tool to understand the replication cycle of MNV.
Animals ; Cell Culture Techniques ; Cycloheximide ; Genome ; Humans ; Mice ; Norovirus ; Polymerase Chain Reaction ; Reverse Transcription ; RNA ; Viruses