• Title/Summary/Keyword: Semi-nested PCR

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Exacerbation of Chronic Gastritis by Mycoplasma Infection through Inducing Inflammation

  • Kang, Kyung-Ah;Kwon, Hyuk-Joon;Hyun, Jin-Won
    • Proceedings of the Korean Society of Applied Pharmacology
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    • 2003.11a
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    • pp.95-95
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    • 2003
  • Mycoplasmas resemble H. pylori in production of ammonia and induction of inflammatory cytokines from immune and non-immne cells. In Republic of Korea infection rate of H. pylori is relatively high but only a proportion of them invite additional inflammation and progress into gastric cancers. Therefore, additional risk factors cannot be excluded. The presence and identification of mycoplasma were confirmed by semi -nested PCR and sequencing and the results were compared with pathological data. Fifty-six samples collected from Korean chronic gastritis patients were used for the study. Twenty-three (41.1%) were positive to mycoplasmas and all of them were identified as human mycoplasmas, M. faucium, M. fermentans, M. orale, M. salivarium and M. spermatophilum. Mycoplasma-infected chronic gastritis samples showed more severe, additional infiltration of neutrophils than non-infected samples and the difference was significant (P < 0.05). In conclusion human mycoplasma infection may playa role in progression of chronic gastritis to metaplasia by inducing additional inflammation.

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Recent (2010-2019) foodborne outbreaks caused by viruses in the Republic of Korea along with their detection and inactivation methods (바이러스에 의한 최근(2010-2019) 국내 식중독 사고와 검출법 및 제어법에 대한 동향 조사)

  • Kwon, Seung-Wook;Kim, Sang-Soon
    • Korean Journal of Food Science and Technology
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    • v.53 no.1
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    • pp.1-11
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    • 2021
  • In this review, recent foodborne outbreaks caused by viruses in the Republic of Korea (2010-2019) were analyzed. The human norovirus was found to be the major foodborne virus causing an average of 94.9% of the viral outbreaks. Reverse-transcription polymerase chain reaction (PCR) with electrophoresis has been widely used to detect viruses, but several rapid detection methods, including real-time PCR, multiplex PCR, and quantum dot assay, have also been suggested. For norovirus inactivation studies, surrogates such as murine norovirus and feline calicivirus have been widely used to identify the reduction rate owing to the limitations in laboratory cultivation. Conversely, direct cell infection studies have been conducted for other foodborne viruses such as adenovirus, astrovirus, rotavirus, and hepatitis A or E virus. Moreover, virucidal mechanisms using various physical and chemical treatments have been revealed. These recent studies suggest that rapid in situ detection and effective control are valuable for ensuring food safety against viral infections.

Prevalence of Toxoplasma gondii in Dogs in Zhanjiang, Southern China

  • Jiang, Hai-Hai;Li, Ming-Wei;Xu, Min-Jun;Cong, Wei;Zhu, Xing-Quan
    • Parasites, Hosts and Diseases
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    • v.53 no.4
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    • pp.493-496
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    • 2015
  • Toxoplasmosis, caused by Toxoplasma gondii, is a parasitic zoonosis with worldwide distribution. The present study investigated the prevalence of T. gondii in dogs in Zhanjiang city, southern China, using both serological and molecular detection. A total of 364 serum samples and 432 liver tissue samples were collected from the slaughter house between December 2012 and January 2013 and were examined for T. gondii IgG antibody by ELISA and T. gondii DNA by semi-nested PCR based on B1 gene, respectively. The overall seroprevalence of T. gondii IgG antibody was 51.9%, and T. gondii DNA was detected in 37 of 432 (8.6%) liver tissue samples. These positive DNA samples were analyzed by PCR-RFLP at 3'- and 5'-SAG2. Only 8 samples gave the PCR-RFLP data, and they were all classified as type I, which may suggest that the T. gondii isolates from dogs in Zhanjiang city may represent type I or type I variant. This study revealed the high prevalence of T. gondii infection in dogs in Zhanjiang city, southern China. Integrated measures should be taken to prevent and control toxoplasmosis in dogs in this area for public health concern.

Pathgenicity on Ginseng and Sequence Assays of Ilyonectria radicicola Isolated from Chestnut Rhizosphere Soils (밤나무 근권토양에서 분리한 Ilyonectria radicicola 균주의 인삼에 대한 병원성 및 유전적 분석)

  • Seo, Mun Won;Song, Jeong Young;Kim, Sun Ick;Oh, Sang Keun;Kim, Hong Gi
    • Korean Journal of Medicinal Crop Science
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    • v.26 no.4
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    • pp.302-307
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    • 2018
  • Background: A soil-borne pathogenic fungus, Ilyonectria radicicola (Cylindrocarpon destructans) causes root rot on ginseng (Panax ginseng C. A. Meyer) and is known to attack many other plants. The Nectria/Neonectria radicicola complex has been renamed as the I. radicicola complex after analysis of its multi-gene relatedness and morphological characteristics. The fungi in this complex have been reclassified into 16 species under the genus Ilyonectria based on characteristics analysis Methods and Results: To obtain useful data from the Korean ginseng root rot, I. radicicola was isolated from the rhizosphere soils of the chestnut tree. They were identified through a pathogenicity test and a survey of the morphological features. The existence of I. radicicola in soil samples was confirmed by PCR detections using nested PCR with species-specific primer sets. These were subsequenctly isolated on semi-selective media from PCR-positive soils. Genetic analysis of the I. radicicola complex containing these pathogens was done by comparing the DNA sequences of the histone h3 region. These isolates originating from the rhizosphere soils of chestnut constituted a clade with other closely related species or I. radicicola isolates originating from ginseng or other host plants, respectively. Additionally, the pathogenicity tests to analyze the characteristics of these I. radicicola isolates revealed that they caused weakly virulent root rot on ginseng. Conclusions: This is the first study reporting that I. radicicola isolates from chestnut rhizosphere soils can attack ginseng plant in Korea. Thus, these results are expected to provide informations in the selection of suitable fields for ginseng cultivation.

Aberrant Expression of E-cadherin in Lung Tissues of Patients with Probable Lung Cancer

  • Yuan, Yu-Lin;Wang, Yu-Ming;Liu, Hua;Qin, Gui-Fang;Tang, Ai-Guo;Duan, Yong
    • Asian Pacific Journal of Cancer Prevention
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    • v.13 no.10
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    • pp.5149-5153
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    • 2012
  • Introduction: This study assessed the relationship of E-cadherin mRNA and protein expression with the diagnosis of lung cancer with the aim of providing an auxiliary diagnostic method. Methods: Semi-quantitative nested RT-PCR and western blotting were applied to detect E-cadherin mRNA transcripts and protein, respectively, in 30 cases of diagnostic lung cancer, 30 cases of clinically suspected patients with lung cancer and 30 cases of other disease. Immunohistochemical staining was also used to detect E-cadherin. Results: Remarkably decreased levels of relative E-cadherin mRNA value and increased E-cadherin protein negativity were observed in probable lung cancer, when compared with possible lung cancer and others. With a threshold of 1.45, relative E-cadherin mRNA value showed a sensitivity of 90% and a specifity of 83% for the diagnosis of lung cancer. The combination of decreased relative E-cadherin mRNA value and negative E-cadherin protein increased the specificity and sensitivity. Conclusion: These data suggest that Chinese patients with diagnostic lung cancer have similar decreased levels of relative E-cadherin mRNA and E-cadherin protein value in the lung cancer tissues as in lung cancer patients in other countries. Measurement of relative E-cadherin mRNA and protein values in lung cancer tissues has potential for lung cancer diagnosis.

Prevalence and Genotypes of Transfusion-Transmitted Virus in Children with Hepatitis and Normal Control (정상 소아와 간염 환자에서 Transfusion-Transmitted Virus의 감염상태와 유전자형)

  • Chung, Ju Young;Han, Tae Hee;Hwang, Eung Soo;Ko, Jae Sung;Seo, Jeong Kee
    • Pediatric Gastroenterology, Hepatology & Nutrition
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    • v.8 no.2
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    • pp.202-212
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    • 2005
  • Purpose: Transfusion transmitted virus (TTV) is a newly discovered virus and to date the contribution of TTV to liver disease remains unclear. Little is known about the frequency of TTV infection in children in Korea. The purpose of this study was to investigate the prevalence and genotypic distribution of TTV carried by healthy children and patients with hepatitis in Korea. Methods: Eighty eight of healthy children and three groups of patients with hepatitis-14 patients with chronic hepatitis B, 12 patients with chronic hepatitis C and 25 patients with hepatitis of unknown etiology-were tested. TTV DNA was detected by semi-nested PCR using primer sets generated from N-22 region and from 5' noncoding region (NCR) of the viral genome. PCR products derived from 8 patients with hepatitis and from 11 healthy children were sequenced and a phylogenetic tree was constructed. Results: TTV was found by PCR with N22 primer in 11.3% of healthy children, 28.5% of children with hepatitis B, 25% of children with hepatitis C, 24% of children with hepatitis of unknown etiology. TTV DNA was found by PCR with 5'NCR primer in 32.9% of healthy children, 71.4% of patients with chronic hepatitis B, in 50% of patients with hepatitis C and in 48% of patients with hepatitis of unknown etiology. TLMV DNA was found in 48.9% of healthy children, 21.4% of patients with hepatitis B, 16.6% of patients with hepatitis C, 40% of patients with hepatitis of unknown etiology. Among the sequenced isolates, 10(52%) belonged to genotype 1 (G1) and others belonged to genotype 2 (G2) or genotype 3 (G3). Among the G1 sequences, 7 were grouped as G1a. Conclusion: TTV infection was common in healthy children and in patients with hepatitis. But, the prevalence of TTV DNA by 5'NCR primer was relatively high in patients with hepatitis B and there may be some association between TTV and hepatitis B virus infection. G1 was the major genotype of the studied population.

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Study on Norovirus Genotypes in Busan, Korea (부산지역에서 분리된 norovirus 유전자형 연구)

  • Kim, Nam-Ho;Park, Eun-Hee;Park, Yon-Koung;Min, Sang-Kee;Jin, Seong-Hyeon;Park, So-Hyun
    • Journal of Life Science
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    • v.21 no.6
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    • pp.845-850
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    • 2011
  • Norovirus (NoV) causes major acute non-bacterial gastroenteritis in humans. NoV genus is a member of the family Caliciviridae, which is transmitted by contaminated food and water or from human to human. Many genotypes of genogroups I and II have been reported because of their high genetic diversity. To obtain molecular epidemiological information on gastroenteritis sporadic cases in Busan, Korea, we analyzed the nucleotide sequences of NoV strains detected during 2008~2010. We performed one step RT-PCR amplifying the open reading frame (ORF) 2 (capsid region) followed by semi-nested PCR. Fecal samples were collected from 4,071 acute gastroenteritis, and genotypes of the 421 positive samples were determined by sequence analysis. Based on partial sequence of capsid region, 7 NoV were categorized into genogroup I and 13 into genogroup II. Prevalent genotypes among gastroenteritis patients within Busan were GII.4, GI.6, GII.5 in 2008~2010. The results of this study will contribute to the currently available epidemiological data and improve public health and hygiene via development of diagnostic methods and sustainable surveillance.

Epidemiology and Clinical manifestations of Enterovirus in Pediatric Inpatient in Incheon (인천지역 소아 입원 환자의 엔테로바이러스 역학과 혈청형에 따른 임상양상 비교)

  • Cho, Byoung Wook;Kwon, Seong Eun;Kwon, Mun Ju;Hur, Myong Je;Kim, Kyung Seon;Hong, Young Jin;Kim, Soon Ki;Kwon, Young Se;Kim, Dong Hyun
    • Pediatric Infection and Vaccine
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    • v.23 no.1
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    • pp.46-53
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    • 2016
  • Purpose: Enterovirus (EV) infection in children can manifest various diseases from asymptomatic infection to nonspecific febrile illness, hand-foot-mouth disease, and aseptic meningitis. This study was aimed to investigate epidemiology and clinical significance of various genotypes of EV infections in pediatric inpatient. Methods: We collected the stool samples from the admitted pediatric patients in Inha University Hospital from March 2014 to March 2015. EV detection and genotype identification were performed by real-time RT-PCR and semi-nested RT-PCR. Phylogenetic trees were constructed by neighbor joining method. Results: A total of 400 samples were collected during study period and 112 patients (28%) were diagnosed with EV infections. The mean age of EV positive patients was 2.66 years (0.1-14) and sex ratio was 1.73:1. Genetic sequences of EVs were identified; coxsackievirus B5 (17, 15.2%), coxsackievirus A16 (13, 11.6%), enterovirus 71 (10, 8.9%), and coxsackievirus A2 (9, 8.0%). Nonspecific febrile illness (96, 86%) was the most common clinical manifestation and the duration of fever was 0-11 days (mean 3.1 days). Rash (44, 39%) and meningitis (43, 38%) were followed. Patients who were attending daycare center or had siblings accounted for 82.1%. Phylogenetic relationship tree revealed 6 distinct genogroups among 56 types of EVs. Conclusions: This study is the report of epidemiology, serotype distribution and clinical manifestations of children with EV infection in Incheon. This data will be helpful for further study about the epidemiology of EV infection in Korea.

Investigation of Norovirus Occurrence and Influence of Environmental Factors in Food Service Institutions of ChungCheong Area (충청지역 집단급식소의 노로바이러스 실태조사와 환경요인의 영향)

  • Jung, Woo-Young;Eom, Joon-Ho;Kim, Byeong-Jo;Yun, Min-Ho;Ju, In-Sun;Kim, Chang-Soo;Kim, Mi-Ra;Byun, Jung-A;Park, You-Gyoung;Son, Sang-Hyuck;Lee, Eun-Mi;Jung, Rae-Seok;Na, Mi-Ae;Yuk, Dong-Yeon;Gang, Ji-Yeon;Heo, Ok-Sun
    • Journal of Food Hygiene and Safety
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    • v.25 no.2
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    • pp.153-161
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    • 2010
  • The purpose of this study was to examine the appearance of norovirus in the water for food in food service institutions and the influence of physicochemical and microbial factors of norovirus in order to work out basic data to predict the detection of norovirus. Among 82 samples of water for food in food service institutions, norovirus appeared in 7 samples and the rate of appearance was 8.5%. As for the type of norovirus, one samples contained GI type (genotype GI-6) and six samples contained GII type (genotype GII-2, GII-4, GII-12). In the regression model of prediction of norovirus, the rate of appearance was correlated with $NH_3$-N, total solids and the consumption of $KMnO_4$, out of such variables as $NH_3$-N, total solids, the consumption of $KMnO_4$, depth, chloride and total colony counts, and its contribution rate for effectiveness was 78.60%. In order to examine the influential factor of environment upon the detection of norovirus, Pearson's correlation analysis was carried out. The predictable regression formula for appearance rate of norovirus was expressed as -1.818 + 42.677 [$NH_3$-N] + 0.023 [total solids] + 0.762 [consumption of $KMnO_4$] -0.009 [depth] -0.146 [chloride] + 0.007 [total colony counts] (R = 0.904, $R^2$ = 0.818, adjusted $R^2$ = 0.786, p < 0.05). The most influential factors upon the detection of norovirus were $NH_3$-N, total solids and the consumption of $KMnO_4$. In other words, when the measured values of $NH_3$-N, total solids and the consumption of $KMnO_4$ were higher, the possibility of appearance of norovirus increased.