• 제목/요약/키워드: Real time polymerase chain reaction

검색결과 824건 처리시간 0.023초

Plasma Real Time-Quantitative Polymerase Chain Reaction of Epstein-Barr Virus in Immunocompetent Patients with Hepatitis

  • Hong, Ji-Hye;Bae, Yon-Jung;Sohn, Joon-Hyung;Ye, Byung-Il;Chun, Jin-Kyong;Kim, Hwang-Min
    • Pediatric Gastroenterology, Hepatology & Nutrition
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    • 제15권1호
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    • pp.38-43
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    • 2012
  • Purpose: Epstein-Barr virus (EBV) hepatitis is a usually asymptomatic and self-limiting disease in immunocompetent patients. However, the range of severity is wide, and the serological diagnosis is typically difficult until the convalescent phase. Thus, we examined the value of plasma EBV DNA real-time quantitative polymerase chain reaction (RT-qPCR) in EBV hepatitis for the timely diagnosis and the relationship between EBV viral load and clinical severity. Methods: Sixty samples were confirmed as having EBV infection by RT-qPCR with the EBV BALF5 gene sequence. We examined the clinical characteristics of EBV hepatitis by reviewing medical records. Results: The median total duration of fever was 8 days (range: 0-13 days). The mean peak value of aspartate aminotransferase (AST) was $241{\pm}214$ U/L, and the mean peak value of alanine aminotransferase (ALT) was $298{\pm}312$ U/L. There was no correlation between the serum levels of liver enzyme and plasma EBV DNA titer ($p$=0.1) or between median total duration of fever and EBV DNA titer ($p$=0.056). The median age of the EBV VCA IgM-negative group was lower compared with the EBV VCA IgM-positive group in EBV hepatitis (2 years vs. 6 years, $p$=0.0009). Conclusion: The severity of EBV hepatitis does not correlate with circulating EBV DNA load according to our data. Furthermore, we suggest that plasma EBV PCR may be valuable in young infants in whom the results of serology test for EBV infection commonly are negative.

주거환경 공기 중 호흡기 바이러스의 검출 (Detection of Airborne Respiratory Viruses in Residential Environments)

  • 박근태;문경환;김형태;박찬정;정호철;임영희
    • 한국환경보건학회지
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    • 제37권4호
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    • pp.306-314
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    • 2011
  • Objectives: Respiratory virus infections are the most common disease among all ages in all parts of the world and occur through airborne transmission. The purpose of this study was to detect and quantitate human respiratory viruses in residential environments. Methods: Air samples were collected from the residential space of apartments in the Seoul/Gyeonggi-do area. The samples were collected from indoor and outdoor air. Among respiratory viruses, influenza A virus, influenza B virus, parainfluenza virus, metapneumovirus, respiratory syncytial virus, and adenovirus were investigated by multiplex polymerase chain reaction. Among the virus-positive samples, we performed adenovirus quantification by real-time polymerase chain reaction. Results: Virus detection rates were 44.0%, 3.8%, 3.4%, and 17.3% in spring, summer, autumn, and winter, respectively. The virus detection rate was higher in winter and spring than in summer and autumn. Adenovirus was most commonly detected, followed by influenza A virus and parainfluenza virus. Virus distribution was not significantly different between indoor and outdoor environments. Conclusions: Although virus concentrations were not high in residential environments, residents in houses with detected viruses may have an increased risk of exposure to airborne respiratory viruses, especially in winter and spring.

Evaluation of Intrinsic Bioremediation of Methyl Tert-butyl Ether (MTBE) Contaminated Groundwater

  • Chen, Colin S.;Tien, Chien-Jun;Zhan, Kai-Van
    • 한국지하수토양환경학회지:지하수토양환경
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    • 제19권5호
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    • pp.9-17
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    • 2014
  • This paper reported the use of real-time polymerase chain reaction (PCR), denaturing gradient gel electrophoresis (DGGE), and the culture-based method in the intrinsic bioremediation study at a petroleum contaminated site. The study showed that phenol hydroxylase gene was detected in groundwater contaminated with benzene, toluene, ethylbenzene, xylene isomers (BTEX) and methyl tert-butyl ether (MTBE). This indicated that intrinsic bioremediation occurred at the site. DGGE analyses revealed that the petroleum-hydrocarbon plume caused the variation in microbial communities. MTBE degraders including Pseudomonas sp. NKNU01, Bacillus sp. NKNU01, Klebsiella sp. NKNU01, Enterobacter sp. NKNU01, and Enterobacter sp. NKNU02 were isolated from the contaminated groundwater using the cultured-based method. Among these five strains, Enterobacter sp. NKNU02 is the most effective stain at degrading MTBE without the addition of pentane. The MTBE biodegradation experiment indicated that the isolated bacteria were affected by propane. Biodegradation of MTBE was decreased but not totally inhibited in the mixtures of BTEX. Enterobacter sp. NKNU02 degraded about 60% of MTBE in the bioreactor study. Tert-butyl alcohol (TBA), acetic acid, 2-propanol, and propenoic acid were detected using gas chromatography/mass spectrometry during MTBE degraded by the rest cells of Enterobacter sp. NKNU02. The effectiveness of bioremediation of MTBE was assessed for potential field-scale application.

Expression of vascular endothelial growth factor in oral squamous cell carcinoma

  • Kim, Seok-Kon;Park, Seung-Goo;Kim, Kyung-Wook
    • Journal of the Korean Association of Oral and Maxillofacial Surgeons
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    • 제41권1호
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    • pp.11-18
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    • 2015
  • Objectives: The goal of this study was to determine the correlation of clinicopathological factors and the up-regulation of vascular endothelial growth factor (VEGF) expression in oral squamous cell carcinoma. Materials and Methods: Immunohistochemical staining of VEGF and quantitative real-time polymerase chain reaction (RT-PCR) of VEGF mRNA were performed in 20 specimens from 20 patients with oral squamous cell carcinoma and another 20 specimens from 20 patients with carcinoma in situ as a controlled group. Results: The results were as follows: 1) In immunohistochemical study of poorly differentiated and invasive oral squamous cell carcinoma, high-level staining of VEGF was observed. Significant correlation was observed between immunohistochemical VEGF expression and histologic differentiation, tumor size of specimens (Pearson correlation analysis, significance r>0.6, P<0.05). 2) In VEGF quantitative RT-PCR analysis, progressive cancer showed more VEGF expression than carcinoma in situ. Paired-samples analysis determined the difference of VEGF mRNA expression level between cancer tissue and carcinoma in situ tissue, between T1 and T2-4 (Student's t-test, P<0.05). Conclusion: These findings suggest that up-regulation of VEGF may play a role in the angiogenesis and progression of oral squamous cell carcinoma.

Suitability of denervated muscle flaps as recipient sites for pancreatic islet cell transplantation

  • Park, Jong-Lim;Kim, Taewoon;Kim, Baek-Kyu
    • Archives of Plastic Surgery
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    • 제48권1호
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    • pp.133-143
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    • 2021
  • Background Extensive research has been conducted on islet transplantation as a possible cure for diabetes. Islet transplantation in the liver via the portal vein has shown remarkable results, but numerous other recipient sites are currently being investigated. We aimed to show the effectiveness of using a muscle flap as a recipient site for islet transplantation. Methods Islet cells were harvested from 12 isogenic Lewis rats, and then diabetes was induced in another 12 isogenic Lewis rats by streptozotocin injection. In six rats, 3,000 islets were transplanted into gastrocnemius muscle flaps, and in the other six rats, the same number of islets were transplanted into the gastrocnemius muscle. The transplanted islet cell function between the two groups was compared by means of blood glucose tests, glucose tolerance tests, immunohistochemistry, and real-time reverse transcription polymerase chain reaction. Results In the muscle flap group, blood glucose levels significantly decreased after islet transplantation. Blood glucose levels were significantly different between the two groups at 3 weeks after transplantation. The muscle flap group showed nearly normoglycemic results upon the glucose tolerance test, whereas the muscle group was hyperglycemic. Immunohistochemical evaluation showed positive results against insulin and glucagon in biopsies of both groups, and the islet cell density was higher in the muscle flap group. There were no statistically significant differences between the two groups in real-time reverse transcription polymerase chain reaction results. Conclusions Our results suggest that muscle flaps are promising candidates for islet cell transplantation.

Status and Prospects of PCR Detection Methods for Diagnosing Pathogenic Escherichia coli : A Review

  • Yim, Jin-Hyeok;Seo, Kun-Ho;Chon, Jung-Whan;Jeong, Dongkwan;Song, Kwang-Young
    • Journal of Dairy Science and Biotechnology
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    • 제39권2호
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    • pp.51-62
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    • 2021
  • Escherichia coli are the predominant facultative bacteria found in the gastrointestinal tract of animals and humans. Some strains of E. coli that acquire virulence factors and cause foodborne and waterborne diseases in humans are called pathogenic E. coli and can be divided into five pathotypes according to the virulence mechanism: EAEC, EHEC, EIEC, EPEC, and ETEC. Although selective media have been developed to detect E. coli, distinguishing pathogenic strains from non-pathogenic ones is difficult because of their similar biochemical properties. Therefore, it is very important to find a new and effective diagnostic method to identify pathogenic E. coli. With recent advances in molecular biology and whole genome sequencing, the use of polymerase chain reaction (PCR) is increasing rapidly. In this review paper, we provide an overview of pathogenic E. coli and present a review on PCR detection methods that can be used to diagnose pathogenic E. coli. In addition, the possibility of real-time PCR incorporating IAC is introduced. Consequently, this review paper will contribute to solving the current challenges related to the detection of pathogenic E. coli.

Distribution Analysis of Candida albicans according to Sex and Age in Clinical Specimen Testing for Sexually Transmitted Diseases

  • Jae Eun Choi;Jae-Sik Jeon;Jae Kyung Kim
    • Journal of Microbiology and Biotechnology
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    • 제33권1호
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    • pp.123-126
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    • 2023
  • The prevalence of candidiasis, a contagious disease with high morbidity and mortality, has sharply increased globally over the last two decades. Candida albicans can cause serious infections in patients with weak immunity and in recipients of prolonged antibiotic treatment. Consequently, rapid and accurate identification of species can play an important role in the treatment of candidiasis. Here, we investigated the positive rate and infection trend of C. albicans according to age, specimen type, and sex using multiplex real-time polymerase chain reaction-based testing of samples collected for the diagnosis of sexually transmitted diseases in Korea between 2018 and 2020. When the type of specimen collected was a swab, the positive rate of C. albicans was higher among younger women, and tended to decrease with age. Analysis of swab samples revealed higher positive rates than urinalysis. The reduction trend in positive rates by age was comparable between the overall samples and urine specimens. Among male patients, the positive rate did not differ substantially across the various types of specimens collected. Previous studies have shown a higher prevalence of non-albicans Candida species than C. albicans in clinical specimens, and exclusion of the former from our analysis may be a limitation of this study. However, our findings contribute significantly to the literature because globally, there is a paucity of epidemiological studies using molecular techniques to detect C. albicans in sexually transmitted disease test samples.

False-Negative Results of Real-Time Reverse-Transcriptase Polymerase Chain Reaction for Severe Acute Respiratory Syndrome Coronavirus 2: Role of Deep-Learning-Based CT Diagnosis and Insights from Two Cases

  • Dasheng Li;Dawei Wang;Jianping Dong;Nana Wang;He Huang;Haiwang Xu;Chen Xia
    • Korean Journal of Radiology
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    • 제21권4호
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    • pp.505-508
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    • 2020
  • The epidemic of 2019 novel coronavirus, later named as severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), is still gradually spreading worldwide. The nucleic acid test or genetic sequencing serves as the gold standard method for confirmation of infection, yet several recent studies have reported false-negative results of real-time reverse-transcriptase polymerase chain reaction (rRT-PCR). Here, we report two representative false-negative cases and discuss the supplementary role of clinical data with rRT-PCR, including laboratory examination results and computed tomography features. Coinfection with SARS-COV-2 and other viruses has been discussed as well.

Prevalence of Bordetella bronchiseptica, Mycoplasma felis, and Chlamydia felis using a newly developed triplex real-time polymerase chain reaction assay in Korean cat population

  • Hye-Ryung, Kim;Gyu-Tae, Jeon;Jong-Min, Kim;Ji-Su, Baek;Yeun-Kyung, Shin;Oh-Kyu, Kwon;Hae-Eun, Kang;Ho-Seong, Cho;Doo-Sung, Cheon;Choi-Kyu, Park
    • 한국동물위생학회지
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    • 제45권4호
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    • pp.305-316
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    • 2022
  • Bordetella (B.) bronchiseptica, Mycoplasma (M.) felis, and Chlamydia (C.) felis are considered as main bacterial pathogens of feline upper respiratory tract disease (URTD). In this study, a new triplex quantitative real-time polymerase chain reaction (tqPCR) assay was developed for the rapid and differential detection of these bacteria in a single reaction. The assay specifically amplified three bacterial genes with the detection limit of below 10 copies/reaction. The assay showed high repeatability and reproducibility, with coefficients of intra-assay and inter-assay variation of less than 1%. Based on the diagnostic results of the assay using 94 clinical samples obtained from cats with URTD signs, prevalence of B. bronchiseptica, M. felis, or C. felis was 10.6%, 36.2%, or 6.4%, respectively, indicating that the diagnostic sensitivity was comparable to those of previously reported monoplex qPCR assays. The dual infection rates for B. bronchiseptica and M. felis or M. felis and C. felis was 2.1% or 3.2%, respectively. These results indicated that M. felis has been widely spread, and its co-infection with B. bronchiseptica or M. felis has been frequently occurred in Korean cat population. The developed tqPCR assay will serve as a promising tool for etiological and epidemiological studies of these three bacterial pathogens and the prevalence data obtained in this study will contribute to expanding knowledge about the epidemiology of feline URTD in Korea.