• Title/Summary/Keyword: Blood PCR

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Carbapenemase-Producing Klebsiella oxytoca Detection Using Molecular Methods (분자학적 방법을 이용한 Carbapenemase-Producing Klebsiella oxytoca 검출)

  • Yang, Byoung Seon;Park, Ji Ae
    • Korean Journal of Clinical Laboratory Science
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    • v.51 no.4
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    • pp.428-435
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    • 2019
  • The rapid increase and dissemination of carbapene mases, such as Klebsiella pneumoniae carbapenemase (KPC), has become a major problem within the field of healthcare-related infection. There are few antibiotics to treat carbapenem-resistant Enterobacteriaceae (CRE) infections, so the identification of resistant bacterial mechanisms is critical to initiate infection control and conduct epidemiological research. A rapid and effective method for detecting KPC-producing bacteria is needed to avoid therapeutic failures and introduce measures to prevent and control the dissemination of these multi-resistant bacteria. During the study period, 31 isolates (seven isolates of Acinetobacter spp., six isolates of Morganella morganii, five isolates of Pseudomonas aeruginosa, five isolates of Proteus mirabilis, one isolate of Proteus vulgaris, two isolates of Enterobacter cloacae, one isolate of Enterobacter aerogenes, one isolate of Klebsiella pneumoniae, one isolate of Klebsiella oxytoca, one isolate of Serratia marcescens and one isolate of Escherichia coli) were identified by the VITEK. Gram negative rod bacteria were the most frequently isolated from urine (35.5%), blood (19.4%), sputum (16.1%), pus (9.7%), ascitic fluid (9.7%), tracheal aspirates (6.5%) and bile juice (3.2%). Analysis using the PCR method identified the blaKPC gene in the K. oxytoca1 strain, but the blaIMP, blaVIM and blaOXA-48 genes are not amplified. In conclusion, diagnosis using the PCR method can accurately and quickly diagnose KPC, thus establishing quick preventive measures to prevent the spread of KPC in hospitals.

Genetic Similarity and Diversity in Crucian Carp(Carassius carassius) Populations by Polymerase Chain Reaction-Random Amplified Polymorphic DNAs

  • Yoon, Jong-Man;Kim, Tae-Sun;Kim, Jong-Yeon
    • Proceedings of the Korean Society of Fisheries Technology Conference
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    • 2001.05a
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    • pp.332-333
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    • 2001
  • Genomic DNA was extracted from the blood of the freshwater crucian carp(Carassius carassius) from Kunsan in Korea, representing genetic similarity by polymerase chain reaction amplification of DNA as twelve of arbitrary primers. The electrophoretic analysis of polymerase chain reaction-random amplified polymorphic DNAs(PCR-RADP) products showed the high levels of similarity between different individuals in crucian carp.

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Effects of geagibokrounghwan on superoxide dismutase gene expression in experimental mouse fed cholesterol (계지복령환이 Cholesterol 식이 생쥐의 SOD 유전자발현능에 미치는 영향)

  • Hong Yun-Pyo;Ro Sung-Hyun
    • Herbal Formula Science
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    • v.6 no.1
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    • pp.215-226
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    • 1998
  • Geagibokrounghwan (桂技茯笭湯) has long been used to cure human diseases such as vascular and blood disorders. However, it is still unkown on its action mechanism, physiolosical and biochemical meaning. Thus, many attempts were tried to show the scientific background covering the above mentioned mechanism. The effect of Geagibokrounghwan, which was known to date, as follow; effective circulation of body blood system, proliferation of leucocytes and antioxidative action. In this study, we have applied the Geagibokrounghwan administration and feed to mouse, to see effects on the expression of superoxide dismutase(SOD) mRNA as antioxidative agent and oxygen radical scavenger. Total RNAs includingmRNA have been isolated from liver and white blood cells after mice were fed with cholesterol in high dose. Also, in a separate group, the cholesterol-administrated mice were fed with Geagibokrounghwan to see the effects on SOD transcription. and then reverse transcriptase-polymerase chain reaction (RT-PCR) usion each primer set (SOD-F;GATGAAAGCGGTGT-3'; SOD-R; 5'-CCTGTGGAGTGATT-3') were performed to trace theamounts of mRNA. SOD mRNA was specifically expressed in Geagibokrounghwan-fed mice at 2 weeks after treatment, however, gradually reduced after 4 weeks. These results indicate that Geagibokrounghwan is highly applicable in treatment of the above mentioned human diseases.

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FHIT Gene Expression in Acute Lymphoblastic Leukemia and its Clinical Significance

  • Malak, Camelia A Abdel;Elghanam, Doaa M;Elbossaty, Walaa Fikry
    • Asian Pacific Journal of Cancer Prevention
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    • v.16 no.18
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    • pp.8197-8201
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    • 2016
  • Background: To investigate the expression of the fragile histidine triad (FHIT) gene in acute lymphoblastic leukemia and its clinical significance. Materials and Methods: The level of expressed FHIT mRNA in peripheral blood from 50 patients with acute lymphoblastic leukemia (ALL) and in 50 peripheral blood samples from healthy volunteers was measured via RT-PCR. Correlation analyses between FHIT gene expression and clinical characteristics (gender, age, white blood count, immunophenotype of acute lymphoblastic leukemia and percentage of blast cells) of the patients were performed. Results: The FHIT gene was expressed at $2.49{\pm}7.37$ of ALL patients against $14.4{\pm}17.9$ in the healthy volunteers. The difference in the expression levels between ALL patients and healthy volunteers was statistically significant. The rate of gene expression did not significantly vary with immunophenotype subtypes. Gene expression was also found to be correlated with increase of total leukocyte and decrease in platelets, but not with age, gender, immunophenotyping or percentage of blast cells. Conclusions: FHIT gene expression is low in acute lymphoblastic leukemia and could be a useful marker to monitor minimal residual disease. This gene is also a candidate target for the immunotherapy of acute lymphoblastic leukemia.

Changes in Median Ages at Death from Selected Cancer Types in Relation to HLA-DRB1/DQB1

  • An, Wan-Xin;Fan, Ya-Xin;Liang, Xiao-Hua;Liu, Hui
    • Asian Pacific Journal of Cancer Prevention
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    • v.15 no.10
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    • pp.4125-4128
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    • 2014
  • The median ages at death from cancers between 1985 and 2005 were calculated to demonstrate that inherent anticancer mechanisms may be a common pathway for different cancers. Seventy-eight patients with gastric, liver and lung cancers, were recruited in the solid cancer group. The leukemia group consisted of 31 patients with three main types of leukemia. The controls were 100 healthy individuals. The samples were typed using an HLA-DR/DQ PCR-SSP typing kit. The results showed that the median ages at death from all causes were 64.7 years in 1985 and 70.1 years in 2005. The range of the median ages at death from all cancers was similar to the corresponding value for deaths attributed to all causes. The frequency of $DRB1^*03$ was 9.6% in the solid cancer group and 3.0% in the control group (p<0.05). The frequency of $DRB1^*04$ in the leukemia group were significantly lower than that of the control (p<0.05). $DRB1^*13$ and $DRB1^*06$ frequencies in the leukemia group were significantly higher than those of the controls (p<0.05). It is suggested that inherent anti-cancer mechanisms may be a common pathway for different cancers and are associated with the immune system and HLA.

Linkage Disequilibrium of Dopamine D2 Receptor Gene in the Korean Population

  • Kang, Byung-Yong;Oh, Sang-Duk;Lee, Kang-Oh
    • Toxicological Research
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    • v.20 no.1
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    • pp.49-53
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    • 2004
  • The genetic basis of hypertension is complex, and has been considered to be associated with the dopamine D2 receptor gene (DD2R). Because association studies using the candidate gene approach may provide important clues regarding the pathogenesis of hypertension and establish basis for further study, we performed the association study on the relationship between genetic polymorphisms in the DD2R gene and hypertension in Koreans. Eighty nine patients with hypertension and 86 age-matched subjects with normal blood pressure were enrolled. Genomic DNA was extracted from peripheral blood leukocytes. PCR-RFLP analysis was performed to detect the three polymorphic Taq I sites in the DD2R gene. There were no significant differences in genotype, allele and haplotype distributions of any polymorphisms in the DD2R gene between two groups, respectively (P>0.05), although significant linkage disequilibriums among these polymorphic sites were detected by pair-wise analysis (P<0.05). Therefore, our negative result suggest that the three Taq I RFLPs in the DD2R gene were not significantly associated with hypertension in Koreans.

Total Cholesterol and Alkaline Phosphatase are Increased in D/D Type of Angiotensin Converting Enzyme

  • Kim, Sung-Su
    • Biomedical Science Letters
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    • v.18 no.4
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    • pp.406-412
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    • 2012
  • The polymorphism (insertion, I or deletion, D) of angiotensin converting enzyme (ACE) gene is designated as the presence of a 287 bp Alu repeat. The D/D homozygote carrier is associated with high ACE activity, and this high activity has been implicated with hypertension, coronary artery disease, or diabetic nephropathy. We studied the clinical candidate marker in ACE gene polymorphism using chemical and hematological analysis. The subjects are divided into normotensive and hypertensive groups and ACE genotype in the group was confirmed by PCR method. Chemical analysis was preceded with Hitachi7060, and hematological analysis was performed using Mythic 22. In 116 targeted people, 17 (38.64%) of 44 I/I genotype group are hypertension, 15 (34.09%) in 44 with D/I, but, D/D type in the 28 cases is 15 patients (53.57%) in hypertension. In hypertension group, biochemical analysis (triglyceride, and alkaline phosphatase) and hematological analysis (white blood cell, platelet) are showed high value in D/D genotype of ACE gene. The relationship between hypertension and ACE genotype is the same results as previously reported and we thought that the high laboratory value of white blood cell, platelet, triglycerides, and alkaline phosphatase are also indicator of hypertension in D/D type of ACE.

AXIN2 Polymorphisms, the β-Catenin Destruction Complex Expression Profile and Breast Cancer Susceptibility

  • Aristizabal-Pachon, Andres Felipe;Carvalho, Thais Inacio;Carrara, Helio Humberto;Andrade, Jurandyr;Takahashi, Catarina Satie
    • Asian Pacific Journal of Cancer Prevention
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    • v.16 no.16
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    • pp.7277-7284
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    • 2015
  • Background: The Wnt/${\beta}$-catenin signaling pathway is an important regulator of cellular functions such as proliferation, survival and cell adhesion. Wnt/${\beta}$-catenin signaling is associated with tumor initiation and progression; ${\beta}$-catenin mutations explain only 30% of aberrant signaling found in breast cancer, indicating that other components and/or regulation of the Wnt/${\beta}$-catenin pathway may be involved. Objective: We evaluated AXIN2 rs2240308 and rs151279728 polymorphisms, and expression profiles of ${\beta}$-catenin destruction complex genes in breast cancer patients. Materials and Methods: We collected peripheral blood samples from 102 breast cancer and 102 healthy subjects. The identification of the genetic variation was performed using PCR-RFLPs and DNA sequencing. RT-qPCR was used to determine expression profiles. Results: We found significant association of AXIN2 rs151279728 and rs2240308 polymorphisms with breast cancer risk. Significant increase was observed in AXIN2 level expression in breast cancer patients. Further analyses showed APC, ${\beta}$-catenin, CK1${\alpha}$, GSK3${\beta}$ and PP2A gene expression to be associated to clinic-pathological characteristics. Conclusions: The present study demonstrated, for the first time, that AXIN2 genetic defects and disturbance of ${\beta}$-catenin destruction complex expression may be found in breast cancer patients, providing additional support for roles of Wnt/${\beta}$-catenin pathway dysfunction in breast cancer tumorigenesis. However, the functional consequences of the genetic alterations remain to be determined.

Identification of a Novel Single Nucleotide Polymorphism in Porcine Beta-Defensin-1 Gene

  • Pruthviraj, D.R.;Usha, A.P.;Venkatachalapathy, R.T.
    • Asian-Australasian Journal of Animal Sciences
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    • v.29 no.3
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    • pp.315-320
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    • 2016
  • Porcine beta-defensin-1 (PBD-1) gene plays an important role in the innate immunity of pigs. The peptide encoded by this gene is an antimicrobial peptide that has direct activity against a wide range of microbes. This peptide is involved in the co-creation of an antimicrobial barrier in the oral cavity of pigs. The objective of the present study was to detect polymorphisms, if any, in exon-1 and exon-2 regions of PBD-1 gene in Large White Yorkshire (LWY) and native Ankamali pigs of Kerala, India. Blood samples were collected from 100 pigs and genomic DNA was isolated using phenol chloroform method. The quantity of DNA was assessed in a spectrophotometer and quality by gel electrophoresis. Exon-1 and exon-2 regions of PBD-1 gene were amplified by polymerase chain reaction (PCR) and the products were subjected to single strand conformation polymorphism (SSCP) analysis. Subsequent silver staining of the polyacrylamide gels revealed three unique SSCP banding patterns in each of the two exons. The presence of single nucleotide polymorphisms (SNPs) was confirmed by nucleotide sequencing of the PCR products. A novel SNP was found in the 5'-UTR region of exon-1 and a SNP was detected in the mature peptide coding region of exon-2. In exon-1, the pooled population frequencies of GG, GT, and TT genotypes were 0.67, 0.30, and 0.03, respectively. GG genotype was predominant in both the breeds whereas TT genotype was not detected in LWY breed. Similarly, in exon-2, the pooled population frequencies of AA, AG, and GG genotypes were 0.50, 0.27, and 0.23, respectively. AA genotype was predominant in LWY pigs whereas GG genotype was predominant in native pigs. These results suggest that there exists a considerable genetic variation at PBD-1 locus and further association studies may help in development of a PCR based genotyping test to select pigs with better immunity.

Prevalence of Dirofilaria immitis Infection in Stray Cats by Nested PCR in Korea

  • Park, Hyung-Jin;Lee, Sang-Eun;Lee, Won-Ja;Oh, Jung-Hyun;Maheswaran, Easwaran;Seo, Kyoung-Won;Song, Kun-Ho
    • Parasites, Hosts and Diseases
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    • v.52 no.6
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    • pp.691-694
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    • 2014
  • The purpose of this study was to conduct a survey of Dirofilaria immitis infection among stray cats in Korea using nested PCR. We included 235 stray cats (121 females and 114 males) and evaluated each for the presence of feline heartworm infection. Blood samples were collected from 135 cats in Daejeon, 50 cats in Seoul, and 50 cats from Gyeonggi-do (Province). Of the 235 DNA samples, 14 (6.0%) were positive for D. immitis. The prevalence of infection in male cats (8/114, 7.0%) tended to be higher than that in female cats (6/121, 5.0%), but the difference was not statistically significant. In each location, 8, 2, and 4 cats were positive for infection, respectively, based on DNA testing. No significant differences in the prevalence were observed among the geographic regions, although the rate of infection was higher in Gyeonggi-do (8.0%) than Daejeon (5.9%) and Seoul (4.0%). We submitted 7 of the 14 D. immitis DNA-positive samples for sequencing analysis. All samples corresponded to partial D. immitis cytochrome c oxidase subunit I gene sequences with 99% homology to the D. immitis sequence deposited in GenBank (accession no. FN391553). To the best of our knowledge, this is the first survey using nested PCR to analyze the prevalence of D. immitis in stray cats in Korea.