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

Laccase activity of Pleurotus ostreatus is significantly increased by the addition of apple pomace. Among various conditions, the best concentration of apple pomace and cultivation time for the production of laccase by P. ostreatus was 2.5% and 9 days, respectively. Reverse transcription polymerase chain reaction analyses of laccase isoenzyme genes, including pox1, pox3, pox4, poxc, poxa3, and poxa1b, revealed a clear effect of apple pomace on transcription induction. Our findings reveal that the use of apple pomace can be a model for the valuable addition of similar wastes and for the development of a solid-state fermenter and commercial production of oyster mushroom P. ostreatus.
Fungi* ; Laccase* ; Pleurotus* ; Polymerase Chain Reaction ; Reverse Transcription

Objective. To determine the diagnostic accuracy of self-collected snorted and spit saliva in detecting COVID-19 using RT-PCR (ssRT-PCR) and lateral flow antigen test (ssLFA) versus nasopharyngeal swab RT-PCR (npRT-PCR). Methods. One hundred ninety-seven symptomatic subjects for COVID-19 testing in a tertiary hospital underwent snort-spit saliva self-collection for RT-PCR and antigen testing and nasopharyngeal swab for RT-PCR as reference. Positivity rates, agreement, sensitivity, specificity, and likelihood ratios were estimated. Results. Estimated prevalence of COVID-19 using npRT-PCR was 9% (exact 95% CI of 5.5% - 14.1%). A higher positivity rate of 13% in the ssRT-PCR assay suggested possible higher viral RNA in the snort-spit samples. There was 92.9% agreement between ssRT-PCR and npRT-PCR (exact 95% CI of 88.4% to 96.1%; Cohen’s Kappa of 0.6435). If npRT-PCR will be assumed as reference standard, the estimated Sensitivity was 83.3% (exact 95% CI of 60.8% to 94.2%), Specificity 93.9% (exact 95% CI of 89.3% to 96.5%), Positive predictive value of 57.7% (exact 95% CI of 38.9% to 74.5%), Negative predictive value of 98.2% (exact 95% CI of 95% to 99.4%), positive likelihood ratio of 3.65 (95% CI of 7.37 to 24.9), negative likelihood ratio of 0.178 (95% CI of 0.063 to 0.499). There was 84.84% agreement (95% exact CI of 79.1% to 89.5%; Cohen’s Kappa of 0.2356) between ssLFAvs npRT-PCR, sensitivity of 38.9% (exact 95% CI of 20.3% to 61.4%), specificity of 89.4% (exact 95% CI of 84.1% to 93.1%), PPV of 26.9% (95% CI of 13.7% to 46.1%), NPV of 93.6% (exact 95% CI of 88.8% to 96.4%), LR+ of 3.67 (95% CI of 1.79 - 7.51), LR – of 0.68 (95% CI of 0.47 - 0.99). Conclusion. Our data showed that snort-spit saliva RT-PCR testing had acceptable diagnostic performance characteristics and can potentially be used as an alternative to the standard nasopharyngeal/oropharyngeal swab RT-PCR test for COVID-19 in certain situations. However, our data also showed that snort-spit saliva antigen testing using lateral flow assay did not offer acceptable performance.
Saliva ; SARS-CoV-2 ; Reverse Transcription ; Reverse Transcriptase Polymerase Chain Reaction

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

This study was purposed to investigate the expression of the growth-factor independence 1 (GFI1) in patients with leukemia and its clinical significance. Bone marrow mononuclear cells were obtained from 65 newly diagnosed leukemia patients including 24 acute myeloid leukemia (AML), 18 chronic myelogenous leukemia (CML), 6 acute lymphoblastic leukemia (ALL), 17 blast crisis of chronic myelogenous leukemia and 13 patients with iron deficiency anemia (IDA) were used as controls. The relative expression of gene gfi1 was detected by reverse transcriptase-polymerase chain reaction (RT-PCR) and taqman quantitative real-time reverse transcription polymerase chain reaction (QRT-PCR). The results showed that gene expression of gene gfi1 in leukemia patients was obviously higher than that in controls and the difference was statistically significant (p < 0.01), in which the expression of gene gfi1 in newly diagnosed CML patients was higher than that in newly diagnosed AML, newly diagnosed ALL, CML-BCP patients and the difference was significant (p < 0.01). Expression of gene gfi1 in lymphocytic blast crisis of CML was higher than that in nonlymphocytic blast crisis of CML, and the difference was significant. It is concluded that gene gfi1 may play an important role in leukemia, especially in CML incidence and progression. The high level expression of gene gfi1 may be participate in the development of lymphoma.
DNA-Binding Proteins ; genetics ; Gene Expression ; Humans ; Leukemia ; genetics ; Reverse Transcriptase Polymerase Chain Reaction ; Transcription Factors ; genetics ; Transcription, Genetic ; genetics

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

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

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

PURPOSE: Free cancer cells exfoliated from cancer-invaded serosa contribute to peritoneal dissemination, the most frequent pattern of recurrence in patients with gastric cancer. To detect free cancer cells, CEA and CA19-9 were introduced as the markers of gastric cancer, and many methods, such as cytology, immunoassay, and reverse transcription polymerase chain reaction (RT-PCR), exist for detecting them. The aim of this study is to define the clinical significance of using immunoassay to measure the levels of CEA and CA19-9 in the peritoneal washings in patients with gastric cancer. MATERIALS AND METHODS: The peritoneal washing fluids were obtained from 130 patients with gastric cancer who received a curative gastrectomy, palliative gastrectomy or open and closure. The pCEA and pCA19-9 levels were measured by using immunoassay and cytology. The results were compared with the clinicopathological data. RESULTS: The pCEA and pCA19-9 levels were correlated with tumor invasion, lymph-node metastasis, and stage (P<0.05). CONCLUSION: A correlation was found between elevated pCEA and pCA19-9 levels measured by immunoassay and the TNM stage. Therefore, a combined pCEA and pCA19-9 assay could be a sensitive detector of peritoneal dissemination, as well as a predictor of postoperative prognosis. pCEA and pCA19-9 may also determine the adjuvant management strategy.
Gastrectomy ; Humans ; Immunoassay ; Neoplasm Metastasis ; Polymerase Chain Reaction ; Prognosis ; Recurrence ; Reverse Transcription ; Serous Membrane ; Stomach Neoplasms*

The incidence and distribution of the human rotavirus G types (VP7 associated: G1~G4) and P types (VP4 associated: P[4], P[6], P[8], P[10]) were determined from 89 rotavirus strains isolated from diarrhea patients between 2001 and 2002 using reverse transcription and multiplex polymerase chain reaction. G types were identified from 83 (95.5%) and P types were from 82 (92.1%) strains. The predominant genotypes were P[4]G2 (28.1%) and P[6]G4 (27%) with much lower incidence of genotypes P[10]G1 (1.1%) and P[10]G3 (1.1%). P[9] type was not detected. A significant genotypic shift was observed that P[4] was the most prevalent genotype that accounted for 75% during the spring season of 2001, coinfection with P[4] and P[6] for 17.5%. P[6] increased gradually to account for 53% of the strains analysed in the following 2002 spring season. Mixed G types revealing coinfections G2/G3 and G3/G4 were found at low frequency (2.2%).
Coinfection ; Diarrhea* ; Genotype ; Humans ; Incidence ; Multiplex Polymerase Chain Reaction* ; Reverse Transcription ; Rotavirus* ; Seasons ; Seoul*