• Title/Summary/Keyword: Virus detection

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Diagnostic Significance of Combined Detection of Epstein-Barr Virus Antibodies, VCA/IgA, EA/IgA, Rta/IgG and EBNA1/IgA for Nasopharyngeal Carcinoma

  • Cai, Yong-Lin;Li, Jun;Lu, Ai-Ying;Zheng, Yu-Ming;Zhong, Wei-Ming;Wang, Wei;Gao, Jian-Quan;Zeng, Hong;Cheng, Ji-Ru;Tang, Min-Zhong
    • Asian Pacific Journal of Cancer Prevention
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    • v.15 no.5
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    • pp.2001-2006
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    • 2014
  • The objective of this study was to investigate the diagnostic significance of EBV antibody combined detection for nasopharyngeal carcinoma (NPC) in a high incidence region of southern China. Two hundred and eleven untreated NPC patients, 203 non-NPC ENT patients, and 210 healthy controls were recruited for the study. The titers of VCA/IgA and EA/IgA were assessed by immunoenzyme assay, and the levels of Rta/IgG and EBNA1/IgA were determined by enzyme-linked immunosorbent assay. The levels of VCA/IgA, EA/IgA, Rta/IgG and EBNA1/IgA demonstrated no association with gender or age (p>0.05). The receiver operating characteristic curve and the area under the curve were used to evaluate the diagnostic value. The sensitivity of VCA/IgA (98.1%) and the specificity of EA/IgA (98.5%) were the highest. When a logistic regression model was used to combine the results from multiple antibodies to increase the accuracy, the combination of VCA/IgA+Rta/IgG, whose area under the curve was 0.99, had the highest diagnostic efficiency, and its sensitivity, specificity and Youden index were 94.8%, 98.0% and 0.93 respectively. The data suggest that the combination of VCA/IgA+Rta/IgG may be most suitable for NPC serodiagnosis.

Detection of Mutations to Zidovudine in the pol Gene of Human Immunodeficiency Virus-1 by Direct Sequencing (인면역결핍 바이러스 pol 유전자 염기서열 결정에 의한 지도부딘 (ZDV) 내성 돌연변이의 탐지)

  • Cho, Young-Keol;Lee, Hee-Jung;Sung, Heung-Sup;Kim, Yoo-Kyum;Kim, Young-Bong;Lee, Yong-Jin;Kim, Mi-Jung;Kim, Dae-Ghon;Won, Young-Ho;Cho, Goon-Jae
    • The Journal of Korean Society of Virology
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    • v.29 no.4
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    • pp.271-281
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    • 1999
  • The nested polymerase chain reaction (PCR) assay was used to determine the sequences of reverse transcriptase (RT) codons 41, 67, 70, 210, 215 and 219 of human immunodeficiency virus-1 (HIV-1) pol gene. Template DNA was obtained from uncultured peripheral blood mononuclear cells from 27 Korean HIV-1 infected patients treated with ZDV and Korean red ginseng. The second PCRs were done for 2 separated regions (RT codons $13{\sim}98$ and $152{\sim}259$) with $5\;{\mu}l$ of the first PCR productNucleotide sequences were determined by direct sequencing. In the 27 patients, CD4+ cell count decreased from $230{\pm}117/{\mu}l$ to $152{\pm}162/{\mu}l$ for $46{\pm}26$ months (Mo), and actual duration of ZDV intake was $72{\pm}16$ Mo. In the 16 patients who had been treated with ZDV therapy ${\ge}25$ Mo, the incidences of 70R, 215F/Y, and 41L were 61%, 28% and 22%, respectively and those of 67N, 210W and 219Q were 17%. The incidences of 215F/Y were 6.7% for group ${\le}12$ Mo treatment, 22.7% for group with 13 to 24 Mo, and 27.8% for group ${\ge}25$ Mo. There was no mutation in 9 patients. It might be associated with the interruption of ZDV therapy for more than 6 months in 6 patients. This study shows that the detection of mutation could be useful prognostic marker with other clinical and virological data, and very low mutation rate is dectected compared to overseas reports.

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Effects of extruded pellet and moist pellet feed on health conditions of olive flounder Paralichthys olivaceus (배합사료 및 습사료 투여가 양식 넙치, Paralichthys olivaceus의 건강도에 미치는 영향)

  • Cho, Mi-Cho;Oh, Yun-Kyeong;Park, Shin-Hoo;Lee, Hae-Young;Kang, Yong-Jin;Park, Myoung-Ae
    • Journal of fish pathology
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    • v.22 no.3
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    • pp.283-291
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    • 2009
  • This study was conducted to evaluate the effects of a commercial extruded pellet (EP) and raw fish moist pellet (MP) diet on disease prevalence and serum chemistry of olive flounder Paralichthys olivaceus grown in two commercial-scale aquaculture farms from July to December in 2008. The contents of serum GOT, GPT and glucose in fish fed EP diet (EP group) were higher than those of fish fed the MP diet (MP group). There were no distinct differences in survival rates and mean detection rates of fish pathogens among fish group fed the experimental diets. However, the mean detection rate of fish pathogens in MP group was higher than that of EP group from July to October which are high water temperature season. The dominant pathogens isolated in EP group were Dactylogyrus sp., E. tarda and red sea bream iridovirus (RSIV). On the other hand, Trichodina sp., Streptococcus spp., viral nervous necrosis virus (VNNV) were dominant in MP group.

Single-tube nested reverse transcription-polymerase chain reaction for simultaneous detection of genotyping of porcine reproductive and respiratory syndrome virus without DNA carryover contamination (DNA 교차오염 방지기능이 있는 single-tube nested reverse transcription-polymerase chain reaction을 이용한 돼지생식기호흡기증후군바이러스 유전형 감별진단)

  • Jeong, Pil-Soo;Park, Su-Jin;Kim, Eun-Mi;Park, Ji-Young;Park, Yu-Ri;Kang, Dae-Young;Cha, Hyun-Ouk;Lee, Kyoung-Ki;Kim, Seong-Hee;Park, Choi-Kyu
    • Korean Journal of Veterinary Service
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    • v.39 no.2
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    • pp.107-116
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    • 2016
  • In the study, we developed and evaluated a uracil N-glycosylase (UNG)-supplemented single-tube nested reverse transcription-polymerase chain reaction (UsnRT-PCR) assay that can carried out first-round RT-PCR and second-round nested PCR in a reaction tube without reaction tube opening and can simultaneously detect EU- and NA-PRRSV. The UsnRT-PCR confirmed to have a preventing ability of mis-amplification by contamination of pre-amplified PRRSV DNA from previous UsnRT-PCR. Primer specificities were evaluated with RNAs extracted from 8 viral strains and our results revealed that the primers had a high specificity for both genotypes of PRRSV. The sensitivity of the UsnRT-PCR was 0.1 $TCID_{50}$/0.1 mL for EU- or NA-PRRSV, respectively, which is comparable to that of previously reported real time RT-PCR (RRT-PCR). Clinical evaluation on 110 field samples (60 sera and 50 lung tissues) by the UsnRT-PCR and the RRT-PCR showed that detection rates of the UsnRT-PCR was 70% (77/110), and was relatively higher than that of the RRT-PCR (69.1%, 76/110). The percent positive or negative agreement of the UsnRT-PCR compared to RRT-PCR was 96.1% (73/76) or 90.9% (30/33), showing that the test results of both assays may be different for some clinical samples. Therefore, it is recommend that diagnostic laboratory workers use the two diagnostic assays for the correct diagnosis for the relevant samples in the swine disease diagnostic laboratories. In conclusion, the UsnRT-PCR assay can be applied for the rapid, and reliable diagnosis of PRRSV without concerns about preamplified DNA carryover contamination that can occurred in PCR process in the swine disease diagnostic laboratories.

Molecular epidemiologic trends of norovirus and rotavirus infection and relation with climate factors: Cheonan, Korea, 2010-2019 (노로바이러스 및 로타바이러스 감염의 역학 및 기후요인과의 관계: 천안시, 2010-2019)

  • Oh, Eun Ju;Kim, Jang Mook;Kim, Jae Kyung
    • Journal of Digital Convergence
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    • v.18 no.12
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    • pp.425-434
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    • 2020
  • Background: Viral infection outbreaks are emerging public health concerns. They often exhibit seasonal patterns that could be predicted by the application of big data and bioinformatic analyses. Purpose: The purpose of this study was to identify trends in diarrhea-causing viruses such as rotavirus (Gr.A), norovirus G-I, and norovirus G-II in Cheonan, Korea. The identified related factors of diarrhea-causing viruses may be used to predict their trend and prevent their infections. Method: A retrospective analysis of 4,009 fecal samples from June 2010 to December 2019 was carried out at Dankook University Hospital in Cheonan. Reverse transcription-PCR (RT-PCR) was employed to identify virus strains. Information about seasonal patterns of infection was extracted and compared with local weather data. Results: Out of the 4,009 fecal samples tested using multiplex RT-PCR (mRT-PCR), 985 were positive for infection with Gr.A, G-I, and G-II. Out of these 985 cases, 95.3% (n = 939) were under 10 years of age. Gr.A, G-I, and G-II showed high infection rates in patients under 10 years of age. Student's t-test showed a significant correlation between the detection rate of Gr.A and the relative humidity. The detection rate of G-II significantly correlated with wind-chill temperature. Conclusion: Climate factors differentially modulate rotavirus and norovirus infection patterns. These observations provide novel insights into the seasonal impact on the pathogenesis of Gr.A, G-I, and G-II.

Detection of Serum Hepatitis B Virus DNA According to HBV Markers in Chronic Hepatitis B Liver Disease (만성 B형 간질환에서 간염 B virus 표식자 발현에 따른 DNA의 검출)

  • Lee, Dong-Jun;Choi, Jin-Su;Kim, Joon-Hwan;Lee, Heon-Ju
    • Journal of Yeungnam Medical Science
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    • v.14 no.1
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    • pp.155-167
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    • 1997
  • The identification of serum HBV DNA is very important for the assessment of the disease activity in persistent infection, for the evaluation of the infectivity of an individuals blood. The dot blot, however, has limited sensitivity and sometimes inconsistent with other serological markers and clinical settings. Using the most important recent advance in molecular biology, the polymerase chain reaction(PCR), specific DNA sequences can be amplified more than a million-fold in a few hours and with this technique the detection of the extreme low level of DNA is possible. This study was to determine sensitivity of the PCR for the detection of serum HBV DNA in comparison with dot blot analysis and to investigate the serum HBV DNA status and clinical significance of PCR in patients with chronic HBsAg positive liver disease. The subjects of this study were 17 patients with asymptomatic HBsAg carriers(9 HBeAg positive patients, 8 anti-HBe positive patients), 91 chronic hepatitis B(50 HBeAg positive patients, 41 anti-HBe positive patients), 57 liver cirrhosis(21 HBeAg positive patients, 36 anti-HBe positive patients), 27 hepatocellular carcinoma(10 HBeAg positive patients, 17 anti-HBe positive patients). The results were summerized as following; The detection rates of HBV DNA by dot blot, PCR were 58.9%, 72.2% in HBeAg positive patients, 34.3%, 53.9% in anti-HBe positive patients. The detection rates of HBV DNA by PCR in HBeAg negative patients were 25.0% in asymptomatic HBsAg carriers, 61.0% in chronic hepatitis B, 52.8% in liver cirrhosis, 52.9% in hepatocellular carcinoma. The positive rate for HBV DNA is a significant difference between HBeAg positive and negative asymptomatic HBsAg carriers, but not significantly difference in other groups. In conclusions, this study confirmed that the PCR is much more sensitive than the dot blot analysis in detecting the HBV DNA in the sera of patients with chronic liver disease. The presence of HBV DNA in the serum was detected by PCR with higher sensitivity and it suggested that active viral replication is still going on in most patients with chronic HBsAg positive liver disease irrespective of HBeAg/anti-HBe status, and PCR may be used as a prognostic factor in asymptomatic HBsAg carriers.

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Rapid Detection Method for Human Rotavirus from Vegetables by a Combination of Filtration and Integrated Cell Culture/Real-Time Reverse Transcription PCR (Filtration과 Integrated Cell Culture/Real-Time Reverse Transcription PCR 기법을 이용한 채소류에서 Human Rotavirus 신속 검출)

  • Hyeon, Ji-Yeon;Chon, Jung-Whan;Song, Kwang-Young;Hwang, In-Gyun;Kwak, Hyo-Sun;Lee, Jung-Soo;Kim, Moo-Sang;Lee, Jung-Bok;Seo, Kun-Ho
    • Korean Journal of Microbiology
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    • v.47 no.2
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    • pp.117-123
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    • 2011
  • The purpose of this study was to evaluate and compare different elution and concentration methods for optimization of human rotavirus (HRV) detection method using real-time RT-PCR and cell culture techniques. The leafy vegetable samples (lettuce, Chinese cabbage) were artificially inoculated with HRV. Viruses were extracted from the vegetables by two different elution buffers, buffer A (100 mM Tris-HCl, 50 mM glycine, 3% beef extract, pH 9.5) and buffer B (250 mM Threonine, 300 mM NaCl, pH 9.5), and the extracted viruses were concentrated by filtration and PEG precipitation sequentially. To determine infectivity of the viruses, the viruses recovered from the samples were infected to the MA-104 cells, and integrated cell culture real-time RT-PCR was performed at 1, 48, 72, 96, 120, 144, 168 h post-infection (p.i.). The elution buffer A was more efficient in extracting the virus from the produce samples tested than the buffer B, 29.54% and 18.32% of recoveries, respectively. The sensitivity of real-time RT-PCR method was markedly improved when the virus was concentrated by the filtration method. When the viruses were eluted and concentrated by buffer A and filtration, respectively, the average recovery rate was approximately 51.89%. When the viruses recovered from samples were infected to MA-104 cell, infectious HRV was detected within 48 h p.i. by ICC/real-time RT-PCR, whereas cytopathic effects were not observed until 72 h p.i. The optimized detection method evaluated in this study could be useful for rapid and reliable detection of HRV in fresh produce products and applied for detection of other food-borne viruses.

Seroepidemiology of Hepatitis B Virus Infection in Healthy Korean Adults in Seoul (정상 성인에 있어서의 B형 간염 바이러스 감염에 관한 혈청역학적 연구)

  • Yoo, Keun-Young;Park, Byung-Joo;Ahn, Yoon-Ok
    • Journal of Preventive Medicine and Public Health
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    • v.21 no.1 s.23
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    • pp.89-98
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    • 1988
  • While there have been not a few reports on the seroepidemiological characteristics of hepatitis B virus (HBV) infection in Korea, most of them, however, have had several limitations; operational definition of HBV infection, validity of detection methods of HBV serologic markers, size of the study population, and confirmation of the vaccination history against HBV, etc. In order to avoid such limitations, authors randomly selected 1,495 healthy adults among the 217,511 insured (target population) of Korean Medical Insurance Corporation, living in seoul, and tested HBV serologic markers by RIA method and conducted direct interview to them. Although HBV serologic markers (HBsAg, anti-HBs and anti-HBc) of all the subjects were tested, 392(26.2%) of interview failure cases and 361 vaccinee were excluded from the actual population. Finally, the serologic markers tested of 742 nonvaccinee (study population) only were analysed for the seroepidemiologic observation of the natural infection of HBV. The seroepidemiological characteristics of HBV infection in Korea were as follows ; 1. Point prevalence of HBs antigenemia was 11.7(9.1{\sim}14.3)% in male, which was slightly higher than that of female, 9.5($3.7{\sim}15.3$)%. This level was one of the highest among those of Asian-Pacific countries. Decreasing tendency of HBsAg prevalence alter the age of 50 was observed, which seems to be due to selective attrition of HBV chronic carriers among the healthy adults and/or to the limited-lasting duration of the HBs antigenemia, in part. 2. Point prevalence of anti-HBc(78.8% in male,50.9% in female) was higher than that of anti-HBs(65.2% in male,46.6% in female), respectively. And both of them were higher in male than in female. Increasing tendency of the prevalence of both antibodies was observed by age, which seems to be largely due to recurrent infection in adults and to some cumulative effect, in part, of their relatively longer-lasting duration. 3. The level of HBV infection defined by positive for at least one of the 3 serologic markers of HBV by RIA method was 84.7($81.8{\sim}87.6$)% in male and 61.2($51.9{\sim}70.5$)% in female, which was also one of the highest among those of Asian-Pacific countries. The proportion of susceptible population to HBV infection among healthy adults was 15.3% in male and 38.8% in female. 4. The relative frequency of current or past infection and chronic carrier among HBV infected person was estimated. The currently or past infected was estimated 75.7% in male and 71.8% in female, and chronic carrier state, 13.8% in male and 14.1% in female. The analysis of the geometric mean of the antibody titer in anti-HBs positive sera indicated also to be compatible with the above findings, suggesting that active, even though inapparent, infection of HBV occur so frequently among healthy adults in Korea.

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Molecular Detection and Characterization of Orf Virus from Outbreak of Contagious Pustular Dermatitis in Korean Indigenous Goats (한국 재래 산양의 전염성 농피성 피부병에서 orf virus의 검출과 B2L 유전자를 통한 계통발생학적 분석)

  • Park, Jin-Ho;Kim, Guk-Jong;Choi, Wook;Kim, Eun-Ha;Han, Jae-Chul;Ou, Sung-Guk;Lee, Jon-Hwa;Cho, Mae-Rim;Song, Hee-Jong;Chae, Jun-Seok
    • Journal of Veterinary Clinics
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    • v.21 no.2
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    • pp.102-108
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    • 2004
  • Orf virus (ORFV), a member of genus Parapoxvirus (family-Poxviridae), a causative agent of contagious ecthyma in sheep and goat leading to a condition commonly known as vesicular dermatitis. Recently, twelve goats from Iksan in Jeonbuk province were observed with clinical signs like necrotic vesicular lesions around the mucosa of mouth, nasal cavity, eye, ear, teats, abdomen and groin. Based on these clinical symptoms, contagious ecthyma infection was suspected. The skin scrapping was collected from lesions for isolation of DNA and subsequent PCR amplification of ORFV specific 235 bp region of B2L gene. All of the samples were found positive by PCR analysis. Sequencing and further phylogenetic analysis of the PCR product revealed 100% identity to Japan isolate of ORFV (Okinawa, GenBank accession number AB080769), and showed 99.6% of similarity to New Zealand strain (NZ-2, GenBank accession number U06671). It was concluded that ORFV strain detected in the present study is homologous to Japan isolate and New Zealand strain. The PCR test based on amplification of B2L gene is a highly useful tools for rapid and specific diagnosis of contagious ecthyma.

Independent and Additive Interaction Between Tumor Necrosis Factor β +252 Polymorphisms and Chronic Hepatitis B and C Virus Infection on Risk and Prognosis of Hepatocellular Carcinoma: a Case-Control Study

  • Jeng, Jen-Eing;Wu, Hui-Fang;Tsai, Meng-Feng;Tsai, Huey-Ru;Chuang, Lea-Yea;Lin, Zu-Yau;Hsieh, Min-Yuh;Chen, Shinn-Chern;Chuang, Wan-Lung;Wang, Liang-Yen;Yu, Ming-Lung;Dai, Chia-Yen;Tsai, Jung-Fa
    • Asian Pacific Journal of Cancer Prevention
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    • v.15 no.23
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    • pp.10209-10215
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    • 2015
  • To assess the contribution of tumor necrosis factor $(TNF){\beta}$ +252 polymorphisms to risk and prognosis of hepatocellular carcinoma (HCC), we enrolled 150 pairs of sex- and age-matched patients with HCC, patients with cirrhosis alone, and unrelated healthy controls. $TNF{\beta}$ +252 genotypes were determined by polymerase chain reaction with restriction fragment length polymorphism. Multivariate analysis indicated that $TNF{\beta}$ G/G genotype [odds ratio (OR), 3.64; 95%CI, 1.49-8.91], hepatitis B surface antigen (OR, 16.38; 95%CI, 8.30-32.33), and antibodies to hepatitis C virus (HCV) (OR, 39.11; 95%CI, 14.83-103.14) were independent risk factors for HCC. There was an additive interaction between $TNF{\beta}$ G/G genotype and chronic hepatitis B virus (HBV)/HCV infection (synergy index=1.15). Multivariate analysis indicated that factors associated with $TNF{\beta}$ G/G genotype included cirrhosis with Child-Pugh C (OR, 4.06; 95%CI, 1.34-12.29), thrombocytopenia (OR, 6.55; 95%CI, 1.46-29.43), and higher serum ${\alpha}$-fetoprotein concentration (OR, 2.53; 95%CI, 1.14-5.62). Patients with $TNF{\beta}$ G/G genotype had poor cumulative survival (p=0.005). Cox proportional hazard model indicated that $TNF{\beta}$ G/G genotype was a biomarker for poor HCC survival (hazard ratio, 1.70; 95%CI, 1.07-2.69). In conclusion, there are independent and additive effects between $TNF{\beta}$ G/G genotype and chronic HBV/HCV infection on risk for HCC. It is a biomarker for poor HCC survival. Carriage of this genotype correlates with disease severity and advanced hepatic fibrosis, which may contribute to a higher risk and poor survival of HCC. Chronic HBV/HCV infected subjects with this genotype should receive more intensive surveillance for early detection of HCC.