• Title/Summary/Keyword: specific primer

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Multiplex PCR of Endotracheal Aspirate for the Detection of Pathogens in Ventilator Associated Pneumonia (기계환기폐렴의 원인균 진단에서 인공기도 흡인액을 이용한 Multiplex PCR과 세균배양 결과의 비교)

  • Song, Ju Han;Myung, Soon Chul;Choi, Song Ho;Jeon, Eun Ju;Kang, Hyung Gu;Lee, Hye Min;Cho, Sung Keun;Choi, Jae Chol;Shin, Jong Wook;Park, In Won;Choi, Byoung Whui;Kim, Jae Yeol
    • Tuberculosis and Respiratory Diseases
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    • v.64 no.3
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    • pp.194-199
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    • 2008
  • Background: Early identification of pathogens can improve the prognosis of patients with ventilator associated pneumonia (VAP). In the present study, we evaluated the feasibility of performing multiplex PCR for endotracheal aspirates to detect three important pathogens (P. aeruginosa, K. pneumoniae and MRSA) in patients with VAP. Methods: The endotracheal aspirates of 24 patients were collected within 24 hours of the diagnosis of VAP for performing multiplex PCR. Forward and reverse primers were designed to target the specific site of each pathogen (the oprL gene for P. aeruginosa, 16S rRNA for K. pneumoniae and the mec gene for MRSA). We analyzed the clinical data of the VAP patients, including the culture reports for the endotracheal aspirates. Results: Twenty-four patients (M:F=18:6, mean age=$70{\pm}11$) with VAP were enrolled. Pathogens were isolated from 11 patients (P. aeruginosa in 2, K. pneumoniae in 1, MRSA in 2, other enteric Gram negative bacilli in 3, S. pneumoniae in 2 and mixed infection in 1). Multiplex PCR detected three cases of P.aeruginosa (2 cases coincided with the culture reports) and four cases of K. pneumoniae (1 matched with the culture report). PCR detected two MRSA cases, which did not coincide with the culture reports. Conclusion: Multiplex PCR of the endotracheal aspirate showed some ability to detect Gram negative bacilli, although caution is required when interpreting the results.

Spread of Cyst Nematodes in Highland Chinese Cabbage Field in Gangwon-do (강원도 고랭지배추 재배지에서 씨스트선충의 분포 확산)

  • Kwon, Soon-Bae;Park, Dong-Kwon;Won, Heon-Seop;Moon, Youn-Gi;Lee, Jae-Hong;Kim, Yong-Bog;Choi, Byoung-Gon;Seo, Hyun-Taek;Ko, Hyoung-Rai;Lee, Jae-Kook;Lee, Dong Woon
    • Korean journal of applied entomology
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    • v.57 no.4
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    • pp.339-345
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    • 2018
  • The sugar beet cyst nematode (SBCN), Heterodera schachtii first detected in Taebaek, Gangwon-do in 2011, is one of the major plant parasitic nematodes that cause economic damage to the Chinese cabbage in highland regions. In addition, the distribution of clover cyst nematode (CCN), H. trifolii was confirmed in the highland Chinese cabbage cultivated regions in 2017. In order to investigate the spread of cyst nematodes, this study has been conducted since 2013 in the highland Chinese cabbage cultivation area. In addition, in 2017, the Real-Time PCR technique with the species-specific primer was used to investigate those two cyst nematodes and the soybean cyst nematode (SCN), H. glycines which is known for its distribution in Korea, focusing on the main production regions of highland Chinese cabbage cultivation. The number of infected fields in the Chinese cabbage plantation in highland increased every year to confirm distribution in Taebaek, Samcheok, Jeongseon and Gangneung in 2017, and the cumulative number of infection fields reached 245 by 2017. Of the 41 possible cyst nematode samples for PCR analysis, 61% were CCN, only 9.8% of the SBCN and 29.3% of the SCN were identified. Therefore, some of the previously known SBCN or CCN discoveries are likely to have been infected with SCN. It is believed that the CCN needs to be controlled in the future as CCN have been found to be dominant species in the highland Chinese cabbage plantation regions.

Occurrence and eradication of Plum pox virus on Ornamentals in Korea, 2016-2017 (2016-2017년 국내 핵과류에서의 자두곰보병 발생 및 방제)

  • Kim, Mikyeong;Kim, Gi-Su;Kwak, Hae-Ryun;Kim, Jeong-Eun;Seo, Jang-Kyun;Hong, Seong-Jun;Lee, Gyeong-Jae;Kim, Ju-Hui;Choi, Min-Kyeong;Kim, Byeong-Ryeon;Kim, Ji-Gwang;Han, In-Yeong;Lee, Hyeon-Ju;Won, Heon-Seop;Kang, Hyo-Jung;Han, Jong-Woo;Ko, Suk-Ju;Kim, Hyo-Jeong;Kim, Seung-Han;Lee, Jung-Hywan;Choi, Hong-Soo
    • Research in Plant Disease
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    • v.25 no.1
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    • pp.8-15
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    • 2019
  • Plum pox virus (PPV) is a significant viral disease in Prunus spp. worldwide. A nationwide survey was started in Prunus spp. orchards, since PPV was first detected from peach in Korea, 2015. During 2016-2017, samples were collected from 30,333 trees in 1,985 orchards of stone fruits in 8 provinces and 4 cities, Korea and tested by RT-PCR using specific PPV primer set. As a result, 21 trees including peach (9 trees), Japanese apricot (4 trees), plum (1 tree), apricot (7 trees) in 10 orchards were infected and controlled by eradication program. Amplicons of the expected size (547 bp) were obtained from total RNA of seven peach trees in 2016, and directly sequenced. BLAST analysis revealed the highest nucleotide (NT) identity (99%) with a PPV D isolates (LC331298, LT600782) in Genbank. The seven isolates from shared nt sequence identities of 98 to 100% with one another. Phylogenetic analysis showed the isolates in peach clustered closely with the PPV-D isolates from Korea, Japan, USA, and Canada. This is, to our knowledge, the first report of the presence of PPV in Prunus spp. orchards in Korea, 2016-2017, we hope that our results and efforts will contribute to effective measures for eradication of PPV.

Influence of heading date difference on gene flow from GM to non-GM rices (GM벼에서 non-GM벼로 유전자 이동에 대한 개화기 차이의 영향 분석)

  • Oh, Sung-Dug;Chang, Ancheol;Kim, Boeun;Sohn, Soo-In;Yun, Doh-Won
    • Journal of the Korean Society of International Agriculture
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    • v.30 no.4
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    • pp.347-356
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    • 2018
  • Genetically modified (GM) crops have been increased continuously over the world and concerns about the potential risks of GM crops have also been increasing. Even though GM crops have not been cultivated commercially in Korea, it should be necessary to develop the safety assesment technology for GM crops. In this study, we investigated the influence of heading date difference on gene flow from GM to non-GM rice. In the experimental plot design, The PAC GM rice was placed in the center as a pollen donor and non-GM rice were placed in eight directions as pollen receivers. Five pollen receiver rice cultivars were Unkawng, Daebo, Saegyejinmi, Nakdong-byeo, and Ilmi which had different flowering times. A total of 266,436, 300,237, 305,223, 273,373, and 290,759 seeds were collected from Unkawng, Daebo, Saegyejinmi, Nakdong, and Ilmi, respectively, which were planted around PAC GM rice. The GM${\times}$non-GM hybrids were detected by repeated spraying of herbicide and PAT immunostrip assay. Finally, the hybrids were confirmed by PCR analysis using PAC gene specific primer. The hybrids were found in Nakdong-byeo which had the same heading date with PAC GM rice. The hybridization rate was 0.0007% at Nakdong-byeo plot. All of GM${\times}$non-GM hybrids were located within 2 m distance from the PAC GM rice zone. The physiological elements including rice heading date were found to be important factors to determine GM?rice out crossing rate with GM rice. Consideration should be taken into for many factors like the physiological elements of field heading date of rice cultivars to set up the safety management guideline for prevention of GM rice gene flow.