• Title/Summary/Keyword: Multiplex

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Analysis of Molecular Epidemiological Properties of Staphylococcus aureus Isolates from Domestic Animals and Human Patients by PCR (Polymerase Chain Reaction을 활용한 국내 동물과 사람환자에서 분리한 Staphylococcus aureus 분리주의 분자역학적 특성분석)

  • Woo Yong-Ku;Kim Shin
    • Korean Journal of Microbiology
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    • v.41 no.1
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    • pp.24-37
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    • 2005
  • This study was conducted to analyze the molecular epidemiological properties and to select the most efficient and reliable PCR method on 116 of Staphylococcus aureus (S. aureus) isolates from Korean cattle, black goat, pig, dog, chicken, mouse and also human clinical cases from hospital. The distribution patterns of SSG [species specific genes; coagulase (coa), protein A (spa), nuclease (nuc) and aroA (RsaI) gene] were analyzed by PCR method. Among the SSGs, the nuc-gene was found in all strains $(100\%)$ tested and followed by coa-gene $(87.9\%)$, spa-gene $(91.4\%)$ and aroA-gene $(26.7\%)$, in order. The genetic subtyping by RFLP method was performed on the coa [AluI] and aroA-gene [RsaI] PCR products. The mecA-gene PCR and PCR-RFLP techniques were chosen to detect and verify of MRSA strains. Only the human strains $(12.1\%)$ were detected the positive mecA-gene products (533 bp), which were divided into two specific bands [201 & 332 bp] by HhaI enzyme digestion. On coa-gene and spa-gene typing, coa-gene was typed with ten kinds of genotype and coa-3 type were determined as the most predominant genotype, while spa-gene was divided into eleven kinds of genotype and also spa-7 type were selected the most prevalent genotype based on their genetic variations. On the aroA and coa-gene subtyping by PCR-RFLP, aroA-gene products were discriminated with only seven types of genotype, while coa-gene products were further divided into an eleven genotype, respectively. In comparison of SID values of five PCR based typing methods, the coa-PCR-RFLP (SID0.894) was evaluated the most efficient and reliable tools and followed by coa-PCR (SID0.883) and aroA-PCR-RFLP (SID0.462), in order. In conclusion, we could determined that the coa-PCR-RFLP method was the most suitable genetic analysis tool for S. aureus and MRSA strains from domestic animals and humans.

Polymorphisms of GSTM1 and CYP1A1, and Susceptibility to Primary Lung Cancer in Korean Males (한국인 남성에서 GSTM1과 CYP1A1 유전자 다형성과 원발성폐암의 유전적 감수성)

  • Bae, Nack-Cheon;Lee, Su-Yeon;Chae, Po-Hee;Kang, Kyung-Hee;Kim, Kyung-Rock;Cha, Seung-Ick;Chae, Sang-Chul;Kim, Chang-Ho;Jung, Tae-Hoon;Park, Jae-Yong
    • Tuberculosis and Respiratory Diseases
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    • v.50 no.5
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    • pp.568-578
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    • 2001
  • Background : Lung cancer is frequently cited as an example of a disease caused solely by exposure to environmental carcinogens. However, there is a growing realization that the genetic constitution is also important in determining individual's susceptibility to lung cancer. This genetic susceptibility may result from functional polymorphims of the genes involved in carcinogen metabolism. In this study, the association between GSTM1 and CYP1A1 polymorphisms and the lung cancer risk in Korean males was investigated. Materials and Method : The study population consisted of 153 male lung cancer patients and 143 healthy male controls. The GSTM1 and CYP1A1 genotypes were determined by multiplex PCR and PCR-RFLP analysis. Result : There were no significant differences in the frequency of the GSTM1 null genotype between the cases and the controls. When the cases were categorized by their histologic type, the frequency of the GSTM1 null genotype in the small cell carcinoma group was higher than those of the controls(67.2% vs 55.9%), but the difference was not statistically significant(OR=1.772 ; 95% CI=0.723-4.340). The distribution of the CYP1A1 MspI genotypes among the cases were similar to those among the controls. When the cases were grouped by their histologic type, the ml/m1, ml/m2, m2/m2 genotypes frequencies among the small cell carcinomas(23.0%, 38.5%, and 38.5%, respectively) were significantly different from those of the controls(36.4%, 46.2%, and 7.4%, respectively, p<0.05). When the m1/m1 genotype was used as a reference, the ml/m2 and m2/m2 genotypes were associated with an increased risk for small cell lung cancer(ml/m2 genotype : OR=1.337, 95% CI=0.453-3.947 ; m2/m2 genotype : OR=3.374, 95% CI=1.092-10.421). Conclusion : These results suggest that the GSTM1 and CYP1A1 genotypes may be a genetic determinant of the risk for lung cancer, particlulary small cell carcinoma. Further investigation is needed to confirm these results.

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Application of Multiplex RT-PCR for Simultaneous Identification of Tomato Spotted Wilt Virus and Thrips Species in an Individual Thrips on Chrysanthemum (시설재배 국화에서 총채벌레의 종 동정 및 보독 바이러스 동시 검출을 위한 다중 진단법 적용)

  • Yoon, Ju-Yeon;Yoon, Jung-Beom;Seo, Mi-Hye;Choi, Seung-Kook;Cho, In-Sook;Chung, Bong-Nam;Yang, Chang Yeol;Gangireddygari, Venkata Subba Reddy
    • Research in Plant Disease
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    • v.26 no.4
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    • pp.264-271
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    • 2020
  • We have developed a simultaneous diagnostic method that can identify both the species of thrips and tomato spotted wilt virus (TSWV) that are problematic in chrysanthemum plants. This is a method of amplifying DNA by performing reverse transcription-polymerase chain reaction by simultaneously adding primers specific to TSWV coat protein (N) gene and primers specific to the internal transcribed spacer 2 region of Frankliniella occidentalis and F. intonsa using total nucleic acid extracted from one thrips. The sizes of DNA fragments for TSWV, F. occidentalis, and F. intonsa were 777, 287, and 367 bp, respectively. These results showed species identification of thrips and whether thrips carrying TSWV can be simultaneously confirmed. Further usefulness of the simultaneous diagnostic method was made from greenhouse survey at chrysanthemum greenhouses in Taean (Chungcheongnam-do) and Changwon (Gyeongsangnam-do) to investigate the identification of thrips species and the rate of thrips carrying TSWV. Of thrips collected from the greenhouses, 83.7% thrips was F. occidentalis and 72.9% F. occidentalis carried TSWV in Taean. Similarly, the diagnostic method showed that 92.2% thrips was F. occidentalis and 84.0% F. occidentalis carried TSWV in Changwon. These results confirm that F. occidentalis is a dominant thrips species and the thrips species plays a crucial role in the transmission of TSWV in chrysanthemum plants in the greenhouses. Taken together, this study showed a simple diagnostic method for thrips identification and epidemiological studies of the timing and spread of TSWV through thrips in chrysanthemum greenhouses in South Korea.

Clinical and Epidemiological Characteristics of Common Human Coronaviruses in Children: A Single Center Study, 2015-2019

  • Choi, Youn Young;Kim, Ye Kyung;Choi, Eun Hwa
    • Pediatric Infection and Vaccine
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    • v.28 no.2
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    • pp.101-109
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    • 2021
  • Purpose: Common human coronaviruses (HCoVs) are relatively understudied due to the mild nature of HCoV infection. Given the lack of local epidemiology data on common HCoVs, we aimed to describe clinical and epidemiological characteristics of common HCoVs in children. Methods: Respiratory viral test results from 9,589 respiratory samples from Seoul National University Children's Hospital were analyzed from January 2015 to December 2019. Viral detection was done by the multiplex reverse transcription polymerase chain reaction. Demographics and clinical diagnosis were collected for previously healthy children tested positive for HCoVs. Results: Of the 9,589 samples tested, 1 or more respiratory viruses were detected from 5,017 (52.3%) samples and 463 (4.8%) samples were positive for HCoVs (OC43 2.8%, NL63 1.4%, 229E 0.7%). All 3 types co-circulated during winter months (November to February) with some variation by type. HCoV-OC43 was the most prevalent every winter season. HCoV-NL63 showed alternate peaks in late winter (January to March) and early winter (November to February). HCoV-229E had smaller peaks every other winter. Forty-one percent of HCoV-positive samples were co-detected with additional viruses; human rhinovirus 13.2%, respiratory syncytial virus 13.0%, influenza virus 4.3%. Common clinical diagnosis was upper respiratory tract infection (60.0%) followed by pneumonia (14.8%), croup (8.1%), and bronchiolitis (6.7%). Croup accounted for 17.0% of HCoV-NL63-positive children. Conclusions: This study described clinical and epidemiological characteristics of common HCoVs (OC43, NL63, 229E) in children. Continuing surveillance, perhaps by adding HKU1 in the diagnostic panel can further elucidate the spectrum of common HCoV infections in children.

Diagnostic Evaluation of the BioFire® Meningitis/Encephalitis Panel: A Pilot Study Including Febrile Infants Younger than 90 Days (BioFire® Meningitis/Encephalitis Panel의 진단적 유용성 평가: 90일 미만 발열영아에서의 예비 연구)

  • Kim, Kyung Min;Park, Ji Young;Park, Kyoung Un;Sohn, Young Joo;Choi, Youn Young;Han, Mi Seon;Choi, Eun Hwa
    • Pediatric Infection and Vaccine
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    • v.28 no.2
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    • pp.92-100
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    • 2021
  • Purpose: Rapid detection of etiologic organisms is crucial for initiating appropriate therapy in patients with central nervous system (CNS) infection. This study aimed to evaluate the diagnostic value of the BioFire® Meningitis/Encephalitis (ME) panel in detecting etiologic organisms in cerebrospinal fluid (CSF) samples from febrile infants. Methods: CSF samples from infants aged <90 days who were evaluated for fever were collected between January 2016 and July 2019 at the Seoul National University Children's Hospital. We performed BioFire® ME panel testing of CSF samples that had been used for CSF analysis and conventional tests (bacterial culture, Xpert® enterovirus assay, and herpes simplex virus-1 and -2 polymerase chain reaction) and stored at -70℃ until further use. Results: In total, 72 (24 pathogen-identified and 48 pathogen-unidentified) CSF samples were included. Using BioFire® ME panel testing, 41 (85.4%) of the 48 pathogen-unidentified CSF samples yielded negative results and 22 (91.7%) of the 24 pathogen-identified CSF samples yielded the same results (enterovirus in 19, Streptococcus agalactiae in 2, and Streptococcus pneumoniae in 1) as those obtained using the conventional tests, thereby resulting in an overall agreement of 87.5% (63/72). Six of the 7 pathogen-unidentified samples were positive for human parechovirus (HPeV) via BioFire® ME panel testing. Conclusions: Compared with the currently available etiologic tests for CNS infection, BioFire® ME panel testing demonstrated a high agreement score for pathogen-identified samples and enabled HPeV detection in young infants. The clinical utility and cost-effectiveness of BioFire® ME panel testing in children must be evaluated for its wider application.

Yearly Occurrence of Thrips Infesting Hot Pepper in Greenhouses and Differential Damages of Dominant Thrips (시설 고추재배지 총채벌레 연중 발생 및 주요 총채벌레의 차등 해충성)

  • Kim, Chulyoung;Choi, Duyeol;Lee, Donghyun;Khan, Falguni;Kwon, Gimyon;Ham, Eunhye;Park, Jungjoon;Kil, Eui-Joon;Kim, Yonggyun
    • Korean journal of applied entomology
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    • v.61 no.2
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    • pp.319-330
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    • 2022
  • Andong is a place to culture the great amount of hot pepper (Capsicum annuum L.) in Korea. This study reports a yearly occurrence (March 31~October 25, 2021) of thrips infesting the hot pepper in Andong. Thrips caught to yellow sticky traps were diagnosed by morphological characters and showed two dominant species: Frankliniella occidentalis and F. intonsa. During this period, a total of 107,874 thrips were caught and included F. occidentalis at about 82%, F. intonsa at about 17%, and the other thrips at about 0.3%. There were two main peaks at May~June and at September~October, respectively, in which the total number of thrips was higher in the second peak and most were F. occidentalis. Interestinly, a low level of thrips occurred during July~August was observed and explained by their susceptibility to high temperatures. A laboratory experiment by exposing thrips to high temperatures showed that thrips were susceptible to temperatures higher than 35℃ and not tolerant to 45℃ for 1 h. Indeed, high temperatures higher than 45℃ were recorded in the greenhouses in Andong during July~August. F. occidentalis was more tolerant to the high temperatures than F. intonsa. On the other hand, the thrips showed the highest occurrence peak at July~August in hot pepper-culturing greenhouse in Kangwon, where the average temperatures were mostly lower than those of Andong and no high temperatures higher than 45℃ were recorded during July~August. A viral disease caused by tomato spotted wilt virus (TSWV) was observed in the hot peppers cultured in Andong greenhouses. Multiplex PCR was used to detect the virus along with identification of thrips. With a high record of about 30%, the virus-infected thrips were detected during all the monitoring period. The virulent thrips were identified to be only F. intonsa. These results suggest that F. occidentalis gives a direct damage especially during harvesting period with their high populations while F. intonsa gives indirect damage by transmitting TSWV.

A Case-Control Study on Effects of Genetic Polymorphisms of GSTM1, GSTT1, CYP1A1 and CYP2E1 on Risk of Lung Cancer (GSTM1과 GSTT1, 그리고 CYP1A1, CYP2E1 다형성이 폐암발생에 미치는 영향에 대한 환자-대조군연구)

  • Nan, Hong-Mei;Kang, Jong-Won;Bae, Jang-Whan;Choe, Kang-Hyeon;Lee, Ki-Hyeong;Kim, Seung-Taik;Won, Choong-Hee;Kim, Yong-Min;Kim, Heon
    • Journal of Preventive Medicine and Public Health
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    • v.32 no.2
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    • pp.123-129
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    • 1999
  • Objectives: This study was performed to investigate sweets of genetic polymorphisms of glutathione S-transferase M1 (GSTM1), glutathione S-transferase M1 (GSTT1), cytochrome P450 1A1 (CYP1A1) and cytoehrome P450 2E1 (CYP2E1) on lung cancer development. Methods: Ninety-eight lung cancer patients and 98 age-sex matched non-cancer patients hospitalized in Chungbuk National University Hospital form March 1997 to August 1998, were the subjects of this case-control study. Direct interview was done and genotypes of GSTM1, GSTT1, CYP1A1 and CYP2E1 were investigated using multiplex PCR or PCR-RFLP methods with DNA extracted from venous blood. Effects of the polymorphisms of GSTM1, GSTT1, CYP1A1 and CYP2E1, lifestyle factors including smoking, and their interactions on lung rancor were statistically analyzed. Results: GSTM1 was deleted in 67.01% of the cases and 58.16% of the controls, and the odds ratio(95% CI) was 1.46(0.82-2.62). GSTT1 deletion was 58.76% for the lung cancer patients and 50.00% for the controls[OR:1.43(0.81-2.51)]. The frequencies of lle/lle, lle/Val and Val/Val of the CYP1A1 polymorphisms were 59.18-18%, 35.71%, and 5.10% for the cases, and 52.04%, 45.92%, 2.04% for the controls, respectively. Risk of lung cancer was not associated with polymorphism of CYP1A1 ($x^2trend=0.253$, p-value>0.05). The respective frequency of c1/c1 c1/c2, c2/c2 genotypes for CYP2E1 were 50.00%, 42.86%, 7.14% for the lung cancer patients, and 66.33%, 30.61%, 3.06% for the controls $(x^2trend=5.783,\;p<0.05)$. c2 allele was a significant risk factor for lung cancer. We also observed a significant association of cigarette smoking history with lung cancer risk. The odds ratio(95% Cl) of cigarette smoking was 3.03(1.58-5.81). In multiple logistic analysis including genotypes of GSTM1, GSTT1, CYP1A1 and CYP2E1, and smoking habit, only snaking habit came out to be a significant risk factor for lung cancer. Conclusion: Genetic polymorphisms of GSTM1, GSTT1, CYP1A1 and CYP2E1 are not so strongly associated with lung cancer as lifestyle factors including cigarette smoking.

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