• Title/Summary/Keyword: PCR diagnosis

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A study on the contamination level of pathogenic microorganisms in beef distribution stages (소고기의 유통 단계별 병원성 미생물 오염도에 관한 연구)

  • 박성도;김용환;고바라다;김철희;윤병철;김조균
    • Korean Journal of Veterinary Service
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    • v.25 no.2
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    • pp.117-126
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    • 2002
  • Contamination levels of pathogenic microorganisms in 145 cases of beef, which were distributed in Gwangju province, had been investigated in each distributed stage and also monitored by general bacterial count and E coli count index. General bacterial count of beef from the slaughterhouse was 10$^4$cfu/g less than the level of promotion(10 cfu/$\textrm{cm}^2$) and E coli count index was also under the level of 10$^2$cfu/$\textrm{cm}^2$ recommended level of the ministry of agriculture and forestry. Pathogenic microorganisms were detected from 23.2% of samples in the consumption stage, 12.5% in the slaughtering stage and 5.6% in the transporting and processing stage. Staphylococcus aureus was isolated in the largest number and its ratio was 9.0%, listeria monocytogenes 5.5% and salmonella spp 1.4%. There were no samples that bacteria had been detected dually. E coli O157:H7 and campylobacter jejuni were not isolated. In raw and chilled beef, isolation rate of pathogenic microorganisms were 13.3% and 16.5% each. Especially in raw beef, L monocytogenes was. isolated in 3 samples among 30 cases (10%) and S aureus in one sample (3.3%). According to a scale of meat store, isolation rates of pathogenic microorganisms were different. It was 28.6% in the small-scale meat store and 16.7% in the large-scale meat store each. Four cases (16.7%) of S aureus were isolated in the large-scale meat store and seven cases (20.0%) of L monocytogenes and 2 cases (5.7%) of salmonella spp were isolated in the small-scale meat store. S aureus was isolated in two places among 10 feeding facilities of the elementary school. This result shows that the sanitation of elementary school feeding facilities is so poor and more careful policy consideration is needed. Eleven strains of S aureus isolated showed ${\beta}$-hemolysis on blood agar, 1 strain ${\alpha}$-hemolysis, and 1 strain ${\gamma}$-hemolysis. Isolated strains of L monocytogenes were reconfirmed in 560 bp by PCR. Conclusively, these results show that the sanitary condition in the stages of slaughtering, transportation-processing and consumption influences the degree of pathogenic microorganisms contamination in beef severely It is necessary to apply thoroughly hazard analysis critical control point in a process of beef distribution and also to develop rapid test methods for microorganism diagnosis. This effort is very important for the supply of safe and clean meat from farm to table and helpful for the improvement of public health.

Effect of Acupuncture and Radix Astragali aqua-acupuncture at Synsu(BL23) on transcriptional expression of mouse cytokine IL-6 (신수혈의 침자극과 황기약침이 실험용 생쥐의 면역활성물질인 cytokine의 IL-6 발현에 미치는 영향)

  • Kim Jong-Soo;Sin Sang-Sup;Kim Cheul-Ho;Park Sun-Dong;Park Won-Hwan
    • Journal of Acupuncture Research
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    • v.15 no.2
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    • pp.147-155
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    • 1998
  • Acupuncture and Radix Astragali aqua-acupuncture stimuli have long been used to cure human diseases. However, it still remains to be unkown on its action mechanism, physiolosical and biochemical aspects. Thus, many attempts were made to show the scientific background covering the above mentioned mechanisms. Most recent studies show that these tests improve blood circulatory system and increase leucocyte counts. In this study, we have applied the acupuncture stimuli to mouse Sinsu(BL-23), which is a stimulative point of oriental medicine, to see if cytokine such as IL-6 can be detected. Mice were treated with lipopolysaccharide(LPS) for inflammation induction, and then reverse transcriptase-polymerase chain reaction (RT-PCR) using each primer set was performed to trace the amounts of mRNA. The results are summarized as follows ; 1. IL-6 was not temporarily expressed in normal mice 15 min after the acupuncture was pulled out. But, it started to show a feeble expression at 30 min after the removal of acupuncture and it started to reduce at 1h. after the acupuncture was pulled out 2. IL-6 was specifically expressed in LPS-treated mouse 30 min after the acupuncture was pulled out. The transcriptional expressions of LPS-treated mice were more effective than those of normal mice at 30 min after the removal of acupuncture 3. IL-6 was not temporarily expressed in normal mice 15 min after Radix Astragali aqua-acupuncture. But it expressed most highly at 30 min, and the transcriptional expressions of IL-6 was continued to 3 h. 4. IL-6 was not expressed in all the time after Radix Astragali aqua-acupuncture in LPS-treated mice. Therefore, a follow-up of cytokine IL-6 can be used not only a basis of the effect of acupuncture and Radix Astragali aqua-acupuncture but a diagnosis giude through the immunological action of thats. And, it is suggested that cytokine's expression by Acupuncture and Radix Astragali aqua-acupuncture stimulation should be continuously elucidated.

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Usefulness of RPS4Y Gene on Sex Determination in Human Teeth (사람치아에서 성별감정시 RPS4Y 유전자의 유용성)

  • Yoon, Wang-Ro;Ahn, Jong-Mo;Yoon, Chang-Lyuk
    • Journal of Oral Medicine and Pain
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    • v.33 no.1
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    • pp.59-66
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    • 2008
  • Ribosomal Protein S4Y(RPS4Y) gene is the human sex-linked gene on the Y chromosome. There are a number of reports on the sex determination using RPS4Y gene analysis for prevention and diagnosis in sex-linked disease. Thus RPS4Y gene is a reliable genetic marker for sex determination in forensic medicine. In general, the sex determination of an unidentified body can be achieved based on anatomical characteristics, but sometimes sex determination was considered to be difficult such as pre-adolescent bodies or decomposed, mutilated bodies. In this case, Sex determination using PCR method in human teeth produces good results. Because human teeth have a great structural durability, the DNA well preserved in the teeth. So author isolated nuclear DNA from the 20 human teeth(10 males, 10 females), performed to detect RPS4Y gene by PCR method. Samples were divided four group(10 pulp and 10 dentinal tissue in male, 10 pulp and 10 dentinal tissue in female). It was found that detection of RPS4Y gene for sex determination was possible in all the male pulp tissues and 6 out of 10 male dentinal tissues. But there was not detected in female pulp and dentinal tissues. In the view of this results demonstrates the possibility that detection of RPS4Y gene with other sex chromosome genes from the human teeth is useful to sex determination in forensic medicine.

An Imported Case of Severe Falciparum Malaria with Prolonged Hemolytic Anemia Clinically Mimicking a Coinfection with Babesiosis

  • Na, Young Ju;Chai, Jong-Yil;Jung, Bong-Kwang;Lee, Hyun Jung;Song, Ji Young;Je, Ji Hye;Seo, Ji Hye;Park, Sung Hun;Choi, Ji Seon;Kim, Min Ja
    • Parasites, Hosts and Diseases
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    • v.52 no.6
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    • pp.667-672
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    • 2014
  • While imported falciparum malaria has been increasingly reported in recent years in Korea, clinicians have difficulties in making a clinical diagnosis as well as in having accessibility to effective anti-malarial agents. Here we describe an unusual case of imported falciparum malaria with severe hemolytic anemia lasting over 2 weeks, clinically mimicking a coinfection with babesiosis. A 48-year old Korean man was diagnosed with severe falciparum malaria in France after traveling to the Republic of Benin, West Africa. He received a 1-day course of intravenous artesunate and a 7-day course of Malarone (atovaquone/proguanil) with supportive hemodialysis. Coming back to Korea 5 days after discharge, he was readmitted due to recurrent fever, and further treated with Malarone for 3 days. Both the peripheral blood smears and PCR test were positive for Plasmodium falciparum. However, he had prolonged severe hemolytic anemia (Hb 5.6 g/dl). Therefore, 10 days after the hospitalization, Babesia was considered to be potentially coinfected. A 7-day course of Malarone and azithromycin was empirically started. He became afebrile within 3 days of this babesiosis treatment, and hemolytic anemia profiles began to improve at the completion of the treatment. He has remained stable since his discharge. Unexpectedly, the PCR assays failed to detect DNA of Babesia spp. from blood. In addition, during the retrospective review of the case, the artesunate-induced delayed hemolytic anemia was considered as an alternative cause of the unexplained hemolytic anemia.

Identification of Viral Pathogens for Lower Respiratory Tract Infection in Children at Seoul During Autumn and Winter Seasons of the Year of 2008-2009 (서울의 2008-2009년 가을, 겨울철에 유행한 소아의 하기도 호흡기 바이러스 병원체의 규명)

  • Kim, Ki Hwan;Kim, Ji Hong;Kim, Kyung Hyo;Kang, Chun;Kim, Ki Soon;Chung, Hyang Min;Kim, Dong Soo
    • Pediatric Infection and Vaccine
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    • v.17 no.1
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    • pp.49-55
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    • 2010
  • Purpose : The Purposes of this study are to identify the circulating etiologic viruses of acute lower respiratory tract infection in children and to understand the relation with clinical diagnosis. Methods : We obtained a total of 418 nasopharyngeal aspirates from children admitted for their acute lower respiratory tract infections at three tertiary hospitals in Seoul from September 2008 to March 2009. We performed multiplex RT-PCR to identify 14 etiologic viruses and analyzed their emerging patterns and clinical features. Results : Average age of patients was 16.4 months old and the ratio of male to female was 1.36. Viruses were detected in 56.2% of a total of 418 samples. Respiratory syncytial virus (35%) was the most frequently detected and followed by human rhinovirus (22%), human bocavirus, adenovirus, human metapneumovirus, parainfluenza virus, influenza virus and human coronavirus. Co-infection reached 21.9 % of positive patients. Conclusion : When we manage the patients with acute lower respiratory infectious diseases, we should remind the role of various viral pathogens, which might be circulating by seasons and by local areas.

Pro-(IL-18) and Anti-(IL-10) Inflammatory Promoter Genetic Variants (Intrinsic Factors) with Tobacco Exposure (Extrinsic Factors) May Influence Susceptibility and Severity of Prostate Carcinoma: A Prospective Study

  • Dwivedi, Shailendra;Singh, Sarvesh;Goel, Apul;Khattri, Sanjay;Mandhani, Anil;Sharma, Praveen;Misra, Sanjeev;Pant, Kamlesh Kumar
    • Asian Pacific Journal of Cancer Prevention
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    • v.16 no.8
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    • pp.3173-3181
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    • 2015
  • Background: It has been hypothesized that IL-18 (pro-) and IL-10 (anti-) inflammatory genetic variants at -607 C/A-137G/C and -819C/T,-592C/A, respectively, may generate susceptibility and severity risk with various modes of tobacco exposure in prostate carcinoma (PCa) patients. IL-18 is a pro-inflammatory cytokine expressed on various cells including prostate gland elements, and is a key mediator of immune responses with anti-cancerous properties. IL-10 is an anti-inflammatory cytokine that is associated with tumour malignancy which causes immune escape. Materials and Methods: The present study was conducted with 540 subjects, comprising 269 prostate carcinoma patients and 271 controls. Genotyping was performed by PCR-RFLP and confirmed by real time PCR probe-based methods. Results: The findings indicated that the mutant heterozygous and homozygous genotype CC and GC+CC showed significant negative associations (p=0.01, OR=0.21; 95% CI: 0.08-0.51 and p=0.011, OR=0.43; 95% CI: 0.22-0.81, respectively) thus, less chance to be diagnosed as cancer against GG genotype of tobacco smoking patients. In addition, a heterozygous GC genotype at the same locus of IL-18 pro-inflammatory cytokine may aggravate the severity (OR=2.82; 95%CI 1.09-7.29 :p=001) so that patients are more likely to be diagnosed in advanced stage than with the GG wild homozygous genotype. Our results also illustrated that anti-inflammatory cytokine (IL-10) genetic variants, although showing no significant association with susceptibility to cancer of the prostate, may gave profound effects on severity of the disease, as -819 TC (OR=4.60; 95%CI 1.35-15.73), and -592 AC (OR=5.04; 95%CI 1.08-25.43) of IL-10 in tobacco chewers and combined users (both chewers and smokers) respectively, are associated with diagnosis in more advanced stage than with other variants. Conclusions: We conclude that promoter genetic variants of IL-18 and IL-10 with various modes of tobacco exposure may affect not only susceptibility risk but also severity in prostate cancer.

Expressions of MicroRNA-150 and MicroRNA-424 Targeted to C-reactive Protein in Trophoblast Cell Line (영양막세포에서의 C-reactive protein 조절 microRNA-150과 microRNA-424 발현 분석)

  • Kim, Hee Sung
    • The Journal of the Korea Contents Association
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    • v.19 no.11
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    • pp.375-382
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    • 2019
  • Abnormalities of trophoblast due to early inflammation in pregnancy increase the expression of CRP and affect maternal-fetal interactions, leading to preterm birth and preeclampsia. However, biomarkers related to the regulation of CRP expression have not been found. In this study, miRNA associated with increased expression of CRP was identified and their expression was analyzed to reveal biomarkers involved in the regulation mechanism of trophoblast inflammation through miRNAs. miRNAs that were predicted to regulate CRP gene expression in miRNA databases (mirna, TargetScan, MicroCosm) were screened and HTR-8/SVneo cell lines were treated with LPS (20 ng/mL) to induce inflammatory responses in vitro, with selected miR-7, miR-150, miR-186 and miR-424. The expression was analyzed by qRT-PCR. As a result, expression of CRP was significantly increased in LPS-treated trophoblast (p<0.001) and miR-150 and miR-424 expression were significantly decreased (p<0.001). Thus, miR-150 and miR-424 are involved in the regulation of CRP expression in inflammatory-induced trophoblast and may be useful for the prenatal diagnosis of inflammatory obstetric diseases.

Multiplex Quantitative Real-time Polymerase Chain Reaction Assay for Rapid Detection of Mycobacterium avium subsp. paratuberculosis in Fecal Samples (분변 시료에서 Mycobacterium avium subsp. paratuberculosis 의 빠른 검출을 위한 다중 실시간 중합효소연쇄반응기법의 개발)

  • Han, Jae-Ik;Jung, Young-Hun;Choe, Changyong;Yoo, Jaegyu;Kang, Seog-Jin;Yoo, Hansang;Park, Hongtae;Kwon, Eung-Gi;Cho, Yong-Il
    • Journal of Veterinary Clinics
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    • v.32 no.3
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    • pp.219-223
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    • 2015
  • Mycobacterium avium subsp. paratuberculosis (MAP) causes paratuberculosis or Johne's disease, an intestinal granulomatous infection in domestic and wild animals. The study aimed to develop and evaluate a panel of multiplex quantitative real-time polymerase chain reaction (mqPCR) assay for simultaneous detection of three MAP-specific genes (IS900, F57 and ISMAP02 genes). The analytic sensitivity (i.e., limit of detection, expressed as cells per 1 ml) was 150 for IS900, 1500 for F57, and 50 for ISMAP02. The specificity of the method was determined by testing 152 bovine fecal samples. Based on the test, it showed that the assay simultaneously detected the target genes in short period of time and at lower cost compared to laboratory routine tests. The test agreement between the assay and routine test was 94%. The discrepancy in the results was due to samples that were tested positive by the panel but negative by the routine tests, suggesting that the assay has higher sensitivity than the routine tests. In conclusion, the mqPCR assay could be a rapid and accurate testing tool for investigating paratuberculosis or Johne's disease cases in domestic and wild animals.

Study on Laboratory Diagnosis of the Ebola Virus and Its Current Trends (에볼라 바이러스 진단법과 개발 동향에 관한 고찰 연구)

  • Jeong, Hye Seon;Kang, Yun-Jung
    • Korean Journal of Clinical Laboratory Science
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    • v.47 no.3
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    • pp.105-111
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    • 2015
  • In late December 2013, the Ebola virus emerged from West Africa. The outbreak started in Guinea and rapidly spread to Liberia and Sierra Leone. Initially, the virus is spread to the human population after contact with infected wildlife and then spread person-to-person through direct contact with body fluids such as blood, sweat, urine, semen, and breast milk. The Ebola virus infects endothelial cells, mononuclear phagocytes and hepatocytes. It causes massive damage to internal tissues and organs, such as blood vessels and the liver, and ultimately death. Most tests for the virus RNA rely on a technology called reverse-transcriptase polymerase chain reaction (RT-PCR). While this method is highly sensitive, it is also expensive, requiring skilled scientists, and delicate power supplies. The strip analytical technique (enzyme-linked immunosorbent assay or ELISA) detects antigens or antibodies to the Ebola virus. This test is cheap and does not require electricity or refrigeration. Despite ongoing efforts directed at experimental treatments and vaccine development, current medical work on the Ebola viral disease is largely limited to supportive therapy. Thus, rapid and reliable diagnoses of the Ebola virus are critically important for patient management, infections, prevention, and control measures.

A Retrospective Analysis of Use in Hospitalized Children with Upper Respiratory Tract Infection (상기도 감염으로 입원한 소아환자에서 항생제 사용에 대한 후향적 분석)

  • Jung, Minyoung;Park, Ji Hyun;Oh, Chi Eun
    • Pediatric Infection and Vaccine
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    • v.24 no.2
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    • pp.87-94
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    • 2017
  • Purpose: The inappropriate prescription of antibiotics in children with upper respiratory tract infection (URTI) is common. This study evaluated the factors that influence antibiotics use in hospitalized children with viral URTI confirmed by reverse transcriptase-polymerase chain reaction (RTPCR) assay. Methods: The medical records of admitted patients who performed RT-PCR assay for respiratory virus pathogens from January 2013 to November 2014 were examined. The demographic and clinical features were compared between patients who were administered antibiotics at admission and those who were not. We also investigated differences between children who continued antibiotics and those who stopped antibiotics after a viral pathogen was identified. Results: In the total 393 inpatients, the median age was 23 months (interquartile range, 13 to 41.3 months). Antimicrobial agents were prescribed in 79 patients (20.1%) at admission. Patients with acute otitis media (AOM) had higher rates of antibiotics prescription than those without AOM (48.1% vs. 2.2%, P<0.001), with an adjusted odds ratio of 91.1 (95% confidence interval, 30.5 to 271.7). Level of high-sensitivity C-reactive protein and the proportion of acute rhinosinusitis were also significantly associated with antibiotics use (P<0.001). Among the 44 patients with viruses identified using the RT-PCR method during hospitalization, antibiotic use was continued in 28 patients (63.6%). AOM was statistically associated with continued antibiotic use in the patients (P=0.002). Conclusions: Although the respiratory virus responsible for URTI etiology is identified, clinicians might not discontinue antibiotics if AOM is accompanying. Therefore, careful diagnosis and management of AOM could be a strategy to reduce unjustified antibiotic prescriptions for children with URTI.