• 제목/요약/키워드: multiplex PCR

검색결과 491건 처리시간 0.033초

A comparison of RPLA and PCR for detection of enterotoxins in methicillin-resistant Staphylococcus aureus(MRSA) strains isolated in dogs

  • Park, Son-il;Han, Hong-ryul
    • 대한수의학회지
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    • 제39권4호
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    • pp.806-810
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    • 1999
  • A multiplex-polymerase chain reaction (PCR) assay was used to detect staphylococcal enterotoxin production by 12 strains of Staphylococcus aureus isolated from clinical specimens. To evaluate the efficacy and/or sensitivity of this method, the results were compared to those obtained with the reversed passive latex agglutination kit (SET-RPLA, Denka Seiken, Japan). Of 10 strains positive by PCR were positive by RPLA but two strains, representing high sensitivity of the former method. Enterotoxin B was the most prevalent by the two methods. The kappa index between the two methods was 0.826, indicating a higher agreement and fully reliable for use. These results would suggest that sensitive, inexpensive, and relatively rapid multiplex-PCR technique may be an effective means for the detection of staphylococcal enterotoxin genes as an alternative to traditional methods such as kits or immunological methods, which depend upon the amount of enterotoxin produced.

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Multiplex-PCR을 이용한 병원성 Yersinia enterocolitica의 신속검출 기법에 관한 연구 (Raped Detection Method for Pathogenic Yersinia enterocolitica by a Multiplex-PCR)

  • 이영기;최성민;오수경;신재영;염곤
    • 환경위생공학
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    • 제15권4호
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    • pp.105-113
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    • 2000
  • We have isolated 65 strains(2.0%) of Y. enterocolitica among 3,219 water samples from 380 spring water sites in Seoul from 1994 to 1999. The biochemical characteristics of isolated strains revealed that TSI was A/A, urea, M.R.($37^{\circ}C$), nitrate, motility($37^{\circ}C$), sorbitol, maltose, manitol, arabinose, mannose, trehalose, xylose were positive(100%) and H$_2$S, arginine, lysine, oxidase, citrate, V.P.($37^{\circ}C$), DNase, motility($37^{\circ}C$), dulcitol, adonitol, lactose and raffinose were negative(100%). In in vitro virulence test, positive rate of AAG and CRMOX were 9.2% and 4.6%, respectively. However in the virulence gene detectable gene detectable test by multiplex-PCR using ail, yst, virF genes, 65 strains were all negative, meaning that Y.enterocolitica strains from domestic spring water were not detected for the virulence. Otherwise, mutiplex-PCR which using ail, yst and subgenus-specific primer pair was the best for identifying the virulence of Y. enterocolitica.

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Direct Multiplex Reverse Transcription-Nested PCR Detection of Influenza Viruses Without RNA Purification

  • Song, Man-Ki;Chang, Jun;Hong, Yeong-Jin;Hong, Sung-Hoi;Kim, Suhng-Wook
    • Journal of Microbiology and Biotechnology
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    • 제19권11호
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    • pp.1470-1474
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    • 2009
  • This paper describes the development a of direct multiplex reverse transcription-nested polymerase chain reaction (PCR) method, devised for simultaneous detection and typing of influenza viruses. This method combines the direct reverse transcription reaction without RNA purification with the enhancement of sensitivity and specificity of nested PCR. The method successfully detected three major human influenza viruses: influenza virus A subtype 1 (H1N1) and subtype 3 (H3N2), and influenza B virus (B). The minimum number of virus particles (pfu/ml) necessary for detection in spiked saliva samples was 200 (H1N1), 140 (H3N2), and 4.5 (B). The method's sensitivity and simplicity will be convenient for use in clinical laboratories for the detection and subtyping of influenza and possibly other RNA viruses.

Detection of Eight Different Events of Genetically Modified Maize by Multiplex PCR Method

  • Kim, Jae-Hwan;Song, Hee-Sung;Heo, Mun-Seok;Lee, Woo-Young;Lee, Soon-Ho;Park, Sun-Hee;Park, Hye-Kyung;Kim, Myung-Chul;Kim, Hae-Yeong
    • Food Science and Biotechnology
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    • 제15권1호
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    • pp.148-151
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    • 2006
  • Multiplex PCR was performed to simultaneously detect eight different events of genetically modified (GM) maize. Specific primers were constructed from GA21, T25, TC1507, Mon810, Mon863, Event176, Bt11, and NK603 events of GM maize. Using this PCR method, specific GM maize was monitored in commercialized foods and feed.

충청지역병원에서 분리된 Extended-Spectrum β-Lactamase 생성 대장균과 폐렴간균의 bla 유전형 및 분자역학적 분석 (bla Genotype and Molecular Epidemiological Analysis of Extended-Spectrum β-Lactamase-Producing Escherichia coli and Klebsiella pneumoniae in Chungcheong Regional Hospitals)

  • 육근돌;양병선;박진숙
    • 미생물학회지
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    • 제50권2호
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    • pp.114-118
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    • 2014
  • 충청지역병원에서 ESBL생성 장내세균 122균주를 분리하여 Clinical and Laboratory Standards Institute (CLSI) 방법에 따라 cefotaxime과 cefotaxime/clavulanate를 이용한 combination disk test (CDT)에 의하여 항균제 감수성을 조사하고, 특이 유전자를 대상으로 multiplex PCR을 실시하여 유전형을 검출하였으며 enterobacterial repetitive intergenic consensus (ERIC)-PCR에 의해 분자 역학조사를 실시하였다. ESBL 생성 Escherichia coli 76균주와 Klebsiella pneumoniae 46균주를 CDT로 확인한 결과, ESBL 생성 E. coli의 경우 96.1%가 양성을 나타내었으며 K. pneumoniae의 경우 93.4%가 양성을 나타냈다. Multiplex PCR 결과, E. coli의 경우 CTX-M-2형이 60.5% 양성으로 나타났으며 K. pneumoniae의 경우 VEB-1형이 56.5%가 양성으로 나타났다. ERIC-PCR을 실시한 결과 E coli는 분리지역에 따라 5개의 cluster을 형성하였고 K. pneumoniae는 4개의 cluster을 형성하였다. ESBL생성 장내세균의 유전형은 임상에서 분리한 균주의 감별과 검출에 유용하였으며 ERIC-PCR의 결과는 분리지역에 따라 cluster을 형성하여 지역 간 감염 감시체계 확립에 도움이 될 것으로 사료된다.

국내 맥류에 발생하는 바이러스병 동시진단 방법 (Simultaneous Detection of Barley Virus Diseases in Korea)

  • 이봉춘;배주영;김상민;나지은;최낙중;최만영;박기도
    • 식물병연구
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    • 제23권4호
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    • pp.363-366
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    • 2017
  • 최근 국내 맥류 재배지에서는 대부분 BaMMV, BaYMV, BYDV의 발생이 확인되고 있다. 본 연구에서는 multiplex reverse transcription polymerse chain reaction (mRT-PCR) 방법에 의해 이들 3종류의 바이러스를 동시에 진단하는 방법을 확립하였다. 이들 3종 바이러스의 외피단백질 유전자 정보를 활용하여 각 바이러스에 대한 primer를 제작하였다. mRT-PCR에 사용한 primer는 RT-PCR 반응의 민감도와 특이성에 의해 선발하여 primer 농도와 mRT-PCR의 조건을 설정하였다. 각 바이러스에 대하여 선발된 primer 사용에 의한 mRT-PCR 결과 BaMMV 594 bp, BaYMV 461 bp, BYDV 290 bp의 PCR 산물을 얻을 수 있었다. 본 연구에서 확립된 맥류 바이러스 동시진단방법은 신속하고 특이적인 진단 뿐 아니라 맥류 바이러스병의 전염 등 역학 연구에도 활용 될 것으로 기대된다.