• Title/Summary/Keyword: Marker component

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Rapid Detection of Rifampicin Resistant M. tuberculosis by PCR-SSCP of rpoB Gene in Clinical Specimens (RpoB 유전자 PCR-SSCP법에 의한 임상검체내 Rifampicin 내성 결핵균의 신속진단)

  • Shim, Tae-Sun;Kim, Young-Whan;Lim, Chae-Man;Lee, Sang-Do;Koh, Youn-Suck;Kim, Woo-Sung;Kim, Dong-Soon;Kim, Won-Dong
    • Tuberculosis and Respiratory Diseases
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    • v.44 no.6
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    • pp.1245-1255
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    • 1997
  • Background : Rifampicin(RFP) is a key component of the antituberculous short-course chemotherapy and the RFP resistance is a marker of multi-drug resistant(MDR) tuberculosis. RPoB gene encodes the $\beta$-subunit of RNA polymerase of M. tuberculosis which is the target of RFP. And rpoB gene mutations are the cause of RFP resistance of M. tuberculosis. Although several reports showed that PCR-SSCP would be a rapid diagnostic method for identifying the RFP resistance, there were few reports Performed using direct, clinical specimens. So we Performed PCR-SSCP analysis of rpoB gene of M. tuberculosis in direct, clinical specimens. Methods : 75 clinical specimens were collected from patients at Asan Medical Center from June to August 1996. After PCR of IS 6110 fragments, 43 both AFB smear-positive and IS6110 fragment PCR-positive specimens were evaluated. The RFP susceptibility test was referred to the referral laboratory of the Korean Tuberculosis Institute. DNA was extracted by bead beater method. And heminested PCR was done using 0.1ul(1uCi) [$\alpha-^{32}P$]-dCTP. SSCP analysis was done using non-denaturating MDE gel electrophoresis. Results : The results of PCR of IS6110 fragments of M. tuberculosis were positive in 55(73%) cases of 75 AFB smear-positive clinical specimens. Of the 55 specimens, RFP susceptibility was confirmed in only 43 specimens. Of the 43 AFB smear-positive and IS6110 fragment-positive specimens, 29 were RFP susceptible and 14 were RFP resistant. All the RFP susceptible 29 strains showed the same mobility compared with that of RFP sensitive H37Rv in SSCP analysis of ropB gene. And all the other RFP resistant 13 strains showed the different mobility. In other words they showed 100% identical results between PCR-SSCP analysis and traditional susceptibility test. Conclusion : The PCR-sseP analysis of rpoB gene in direct clinical specimens could be used as a rapid diagnostic method for detecting RFP resistant M. tuberculosis.

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Seasonal Variations of Microphytobenthos in Sediments of the Estuarine Muddy Sandflat of Gwangyang Bay: HPLC Pigment Analysis (광합성색소 분석을 통한 광양만 갯벌 퇴적물 중 저서미세조류의 계절변화)

  • Lee, Yong-Woo;Choi, Eun-Jung;Kim, Young-Sang;Kang, Chang-Keun
    • The Sea:JOURNAL OF THE KOREAN SOCIETY OF OCEANOGRAPHY
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    • v.14 no.1
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    • pp.48-55
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    • 2009
  • Seasonal variations of microalgal biomass and community composition in both the sediment and the seawater were investigated by HPLC pigment analysis in an estuarine muddy sandflat of Gwangyang Bay from January to November 2002. Based on the photosynthetic pigments, fucoxanthin, diadinoxanthin, and diatoxanthin were the most dominant pigments all the year round, indicating that diatoms were the predominant algal groups of both the sediment and the seawater in Gwangyang Bay. The other algal pigments except the diatom-marker pigments showed relatively low concentrations. Microphytobenthic chlorophyll ${\alpha}$ concentrations in the upper layer (0.5 cm) of sediments ranged from 3.44 (March at the middle site of the tidal flat) to 169 (July at the upper site) mg $m^{-2}$, with the annual mean concentrations of $68.4{\pm}45.5,\;21.3{\pm}14.3,\;22.9{\pm}15.6mg\;m^{-2}$ at the upper, middle, and lower tidal sites, respectively. Depth-integrated chlorophyll ${\alpha}$ concentrations in the overlying water column ranged from 1.66 (November) to 11.7 (July) mg $m^{-2}$, with an annual mean of $6.96{\pm}3.04mg\;m^{-2}$. Microphytobenthic biomasses were about 3${\sim}$10 times higher than depth-integrated phytoplankton biomass in the overlying water column. The physical characteristics of this shallow estuarine tidal flat, similarity in taxonomic composition of the phytoplankton and microphytobenthos, and similar seasonal patterns in their biomasses suggest that resuspended microphytobenthos are an important component of phytoplankton biomass in Gwangyang Bay. Therefore, considering the importance of microphytobenthos as possible food source for the estuarine benthic and pelagic consumers, a consistent monitoring work on the behavior of microphytobenthos is needed in the tidal flat ecosystems.