BACKGROUND: Real-time reverse transcriptase PCR (rRT-PCR) is widely used to detect novel influenza A/H1N1. We had observed several cases with positive result for influenza A and negative result for novel influenza A/H1N1 during a novel influenza A/H1N1 outbreak. The causes of those results were investigated in this study. METHODS: A total of 913 cases tested with rRT-PCR for novel influenza A/H1N1 (Real-time Ready Influenza A/H1N1 Detection Set, Roche Diagnostics, Germany) during 25 August 2009 to 8 September 2009 was enrolled in this study. Cases showing positive result for influenza A (M gene) and negative result for novel influenza A/H1N1 (H1 gene) were tested with multiplex RT-PCR for seasonal influenza and novel influenza A/H1N1 (Seeplex FluA ACE Subtyping kit, Seegene, Korea), and the amplicons were directly sequenced. RESULTS: One hundred and eleven cases (12.2%) were positive for novel influenza A/H1N1. Twenty-seven cases (3.0%) were positive for influenza A, but negative for novel influenza A/H1N1. Subtypes of influenza A were determined in 25 cases by multiplex RT-PCR and nucleotides sequencing. One novel influenza A/H1N1, six seasonal influenza A/H1N1, three seasonal influenza A/H3N2, and 15 influenza A/H9N2 were detected. CONCLUSIONS: Subtypes of influenza A were determined in most cases with positive result for influenza A and negative result for novel influenza A/H1N1. Several cases with seasonal influenza A were detected. Even if a nonepidemic period of seasonal influenza, tests for seasonal influenza A can help in the differential diagnosis of influenza.
Diagnosis, Differential ; Influenza, Human ; Nucleotides ; Reverse Transcriptase Polymerase Chain Reaction ; RNA-Directed DNA Polymerase ; Seasons

BACKGROUND: Mycobacterium leprae, the etiological agent of leprosy, is noncultivable on axenic media, so, the viability of M. leprae for clinical or experimental applications is often unknown. Quantitative reverse transcriptase PCR (RT-PCR) assays were recently introduced as the new tools for M. leprae viability determination. OBJECTIVE: For evaluating of correlation of results of quantitative real-time PCR(16S rRNA/RLEP) & AFB stain of slit skin smear & histopathology & estimating the viability of M. leprae, the author studied the comparison of results of them METHODS: Of 46 samples from 27 patients(MB 24 cases, PB 3 cases), M. leprae 16S rRNA was used as RNA targets, and M. leprae repetitive element (RLEP) DNA was used to determine relative bacterial numbers and the viability by the quantitative real-time PCR. The ratio of 16S rRNA and RLEP as the indicator of viability was calculated. Student t test and linear Pearson correlation were done by SPSS. RESULTS: There was a correlation between between 16S rRNA/RLEP ratio and BI (r=0.369, p=0.012), and was statistically significant between 16S rRNA/RLEP ratio and histopathological positivity of AFB (p=0.011). However there was no correlation between 16S rRNA/RLEP ratio and MI. CONCLUSIONS: Although the correlation between between 16S rRNA/RLEP ratio and BI and the statistically significant between 16S rRNA/RLEP ratio and histopathological positivity of AFB, there was no correlation between 16S rRNA/RLEP ratio and MI. It needs the further evaluation the correlation about that.
DNA ; Humans ; Leprosy* ; Mycobacterium leprae ; Real-Time Polymerase Chain Reaction* ; Reverse Transcriptase Polymerase Chain Reaction ; RNA ; Skin*

Hepatitis C virus (HCV) has been identified as an important cause of posttransfusion hepatitis, but vertical transmission of chronic infected HCV RNA positive mothers has been documented in some cases. The reports of the risk of perinatal infection have been widely varied in the literature. The authors experienced one case of vertical transmission of HCV in an infant of a mother who had hepatitis C during pregnancy. At admission, HCV RNA (+), Ig G anti HCV (+) and Ig M anti HCV (+) were found in the mother Also at admission, HCV RNA (+), Ig G anti HCV (+), Ig M anti HCV (+), elevation of liver aminotransferase level and hepatosplenomegaly on ultrasonography were found in the baby on day 31. HCV RNA (-), Ig M anti HVC (-) and normal of liver aminotransferase level were noted on day 250 in the serum of the infant. We used reverse transcriptase polymerase chain reaction (RT-PCR) technique to find a very small amount of HCV RNA in the serum. All the findings suggest vertical transmission of HCV RNA from mother to infant during 3rd trimester of pregnancy.
Hepacivirus* ; Hepatitis C* ; Hepatitis* ; Humans ; Infant* ; Liver ; Mothers* ; Pregnancy* ; Reverse Transcriptase Polymerase Chain Reaction ; RNA ; Ultrasonography

Wounds on fetal skin can be repaired without leaving scars until the second trimester, but after this period, skin wounds leave scars as in adults. It's known that certain growth factors such as TGF-beta, and bFGF are present at a very low levels during wound repair in fetal skin. These low levels of growth factors minimize inflammatory response and fibroblast proliferation at the wound site, which in turn inhibit collagen synthesis, and thus, allows scarless wound healing. Recently bone morphogenetic proteins (BMPs), one of the TGF-beta superfamily members, have been studied in the wound healing process. According to several studies, BMPs are related to the differentiation and growth of epithelial and mesenchymal cells, but the precise functions of BMPs and of BMP receptors on skin wound healing have not been elucidated. In this study, we investigated the expression pattern of BMP receptors in fetal skin during the second trimester and in adult skin by immunohistochemical staining and RT-PCR. BMP receptors were detected on the suprabasal epithelial cells and in the hair follicles in adult skin, but were not defected in the fetal skin except for the hair follicles. This was confirmed by confirming mRNA levels of BMP receptors by RT-PCR in both adult and fetal skins. In conclusion, BMPs and BMP receptors seem to be related to fetal and adult wound healing, and low levels of BMPs and BMP receptors during the second trimester seem to contribute to scarless wound healing in the fetus, as is TGF-beta during the second trimester.
Fetus/metabolism ; Human ; Immunohistochemistry ; Receptors, Cell Surface/*metabolism ; Reverse Transcriptase Polymerase Chain Reaction ; Skin/*embryology/*metabolism

PURPOSE: Viral etiology is common in cases of children with acute diarrhea, and antibiotic therapy is usually not required. Therefore, it is important to determine the distribution of common viruses among children hospitalized with acute diarrhea. METHODS: We included 186 children who suffered from acute diarrhea and were hospitalized at the Wonkwang University Hospital Pediatric ward from December 1, 2010 to June 30, 2011 in this study. Stool samples were collected and multiplex reverse transcriptase polymerase chain reaction (multiplex RT-PCR) was used to simultaneously determine the viral etiology such as rotavirus, norovirus, astrovirus, or adenovirus. RESULTS: Causative viruses were detected in 72 of the 186 cases (38.7%). The mean age of the virus-positive cases was 1 year and 9 months (range, 1 month to 11 years). Rotavirus was detected in 50/186 (26.9%); norovirus, in 18/186 (9.7%); and astrovirus, in 3/186 cases (1.6%). Adenovirus was not detected in any of the cases. Proportions of norovirus genogroups I and II were 21.1% and 78.9%, respectively. Four of the 51 rotavirus-positive cases (7.8%) had received rotavirus vaccination at least once. The mean duration of diarrhea was 2.8 days (range, 1 to 10 days) and vomiting occurred in 39 of the 72 cases (54.2%). CONCLUSION: Viral etiology was confirmed in about one-third of the children with acute diarrhea, and the most common viral agent was rotavirus, followed by norovirus.
Adenoviridae ; Child ; Diarrhea ; Gastroenteritis ; Humans ; Korea ; Norovirus ; Reverse Transcriptase Polymerase Chain Reaction ; Rotavirus ; Vaccination ; Vomiting

We report a case of chronic myelogenous leukemia displaying a variant Philadelphia translocation t(11;22)(q25;q11.2). Breakpoint 11q25 has not previously been reported. Reverse transcriptase polymerase chain reaction and fluorescence in-situ hybridization demonstrated the BCR/ABL rearrangement.
Fluorescence ; Leukemia, Myelogenous, Chronic, BCR-ABL Positive* ; Reverse Transcriptase Polymerase Chain Reaction

PURPOSE: This study was undertaken to investigate whether expression of angiogenic cytokines including IL-8 and MCP-1 is induced in the cultured RPE by oxidative stress. METHODS: ARPE-19 cells were exposed to H2O2 and paraquat for 24, 48, and 72 hours and then assayed for the detection of IL-8 and monocyte chemotactic protein (MCP)-1 expression by reverse transcriptase polymerase chain reaction (RT-PCR) method. To determine whether the antioxidant overcame the oxidative stress, we observed expression of IL-8 and MCP-1 gene after ARPE cells were treated by paraquat and antioxidants including N-acetylcysteine (NAC) and pyrrolidine dithiocarbamate (PDTC) at the same time for 72hr. RESULTS: Expressions of the angiogenic cytokines, IL-8 and MCP-1 were increased by incubating with paraquat for 72hr. Antioxidants including NAC and PDTC decreased the expression of IL-8 and MCP-1 gene by counteracting the paraquat effect. CONCLUSIONS: Oxidative stress stimulates angiogenic cytokines including IL-8 and MCP-1 in the cultured RPE. These results may be significant for understanding the cause of age-related macular degeneration and its treatment.
Acetylcysteine ; Antioxidants ; Cytokines ; Interleukin-8 ; Macular Degeneration ; Monocytes ; Oxidative Stress ; Paraquat ; Reverse Transcriptase Polymerase Chain Reaction

BACKGROUND: Tigecycline resistance has emerged recently and has shown diverse mechanisms. The aim of this study was to assess the role of efflux activity in tigecycline resistance in 120 clinical isolates of A. baumannii using two methods: the H33342 accumulation assay and adeB real-time reverse transcriptase polymerase chain reaction. In addition, we analyzed the correlation between the expression level of adeB and H33342 accumulation level. METHODS: A. baumannii clinical isolates was divided into tigecycline-resistant (49 strains), intermediate (40 strains), and susceptible (31 strains) groups. The H33342 accumulation was measured in the absence or presence of the efflux pump inhibitor carbonyl cyanide 3-chlorophenylhydrazone (CCCP). Real-time RT-PCR was performed to determine the relative expression of the adeB gene in A. baumannii clinical isolates. RESULTS: The level of H33342 accumulation in the resistant group was relatively lower than those in the other groups. The addition of CCCP caused a significantly increased fold change in H33342 accumulation in the tigecycline-resistant group. Significant difference in the fold change level in H33342 accumulation was found between tigecycline-susceptible and resistant isolates. Those findings support the role of efflux pumps of which substrates are H33342 in the resistance of tigecycline. Significant differences in the relative expression levels of adeB were shown between tigecycline-susceptible and resistant groups also. CONCLUSION: The results showed that several efflux pumps of which substrates were H33342 can contribute to tigecycline resistance. The adeB overexpression can also contribute to tigecycline resistance. It is possible that efflux pumps other than adeB efflux pumps contribute to tigecycline resistance because there was no correlation between fold change level in H33342 accumulation and adeB expression level.
Acinetobacter baumannii* ; Acinetobacter* ; Carbonyl Cyanide m-Chlorophenyl Hydrazone ; Reverse Transcriptase Polymerase Chain Reaction

PURPOSE: Rapid antigen test (RAT) is used to screen influenza rapidly. The clinical sensitivity of RAT was poor for 2009 H1N1 influenza. The aim of this study was to identify the correlation of time gap (TG) between fever onset and the sensitivity of RAT for 2009 H1N1 influenza. METHODS: Data were collected retrospectively during the pandemic H1N1 2009 influenza season between October 2009 and February 2010. The RAT was done by using SD Bioline influenza antigen (Standard Diagnostics Inc.) in nasopharyngeal swab. The 2009 H1N1 influenza was confirmed by real-time reverse transcriptase polymerase chain reaction (rRT-PCR). Specimens were categorized according to the TG between fever onset and performance of RAT. They were classified into <24 hours (TG1), 24 to 48 hours (TG2), 48 to 72 hours (TG3), 72 to 96 hours (TG4), 96 to 120 hours (TG5), >120 hours (TG6). RESULTS: The overall sensitivity of RAT was 69.9%. The TG dependent sensitivity of RAT at TG1, TG2, TG3, TG4, TG5, and TG6 was 64.3%, 73.3%, 61.1%, 88.9%, 83.3%, and 61.1% respectively. The sensitivity of RAT was the highest when the TG was 72 to 96 hours. But this result was not statistically significant. CONCLUSION: Correlation of TG between fever onset and the sensitivity of RAT for 2009 H1N1 influenza was not statistically significant. But our study suggested that 72 to 96 hours after fever onset is the most sensitive time of RAT. Timely optimal performance of the RAT could have a significant impact on improving results. Further evaluation for better sensitivity would be needed.
Animals ; Fever ; Influenza, Human ; Pandemics ; Rats ; Retrospective Studies ; Reverse Transcriptase Polymerase Chain Reaction ; Seasons

PURPOSE: Although the 'No-touch' isolation technique was introduced by Turnbull et al. in 1967, the controversy over whether or not it reduces the risk of metastasis during surgery exists even today. The aim of this study was to evaluate the effect of the 'No-touch' isolation technique in primary colorectal cancer surgery. METHODS: The evaluation was done by comparing the levels of CEA and CEA m-RNA expression from the same draining vein before and after tumor mobilization. Blood samples from 25 patients with primary colorectal cancer were collected for analysis. At the time of surgery, the main draining vein from the tumor was isolated and ligated at the proximal end. The 1st blood samples were collected just prior to tumor mobilization, and the 2nd samples right after. Both samples were analyzed for serum CEA level and CEA mRNA expression by using reverse transcriptase polymerase chain reaction (RT-PCR). RESULTS: The mean CEA value from draining veins after tumor mobilization (8.08+/-8.98 ng/ml) was significantly higher than it was before mobilization (4.17+/-4.98 ng/ml). CEA mRNA was detected in 16% (4/25) of the blood specimens post-mobilization, whereas it was detected in only 4% (1/25) of the pre-mobilization samples. CONCLUSIONS: The results suggest the validity of using the 'No-touch' isolation technique to reduce the risk of metastasis into the draining vein during mobilization.
Carcinoembryonic Antigen ; Colorectal Neoplasms ; Humans ; Neoplasm Metastasis ; Reverse Transcriptase Polymerase Chain Reaction ; RNA, Messenger ; Veins