6 research outputs found
Evaluation of molecular technologies by using RNA for tuberculosis diagnosis
์์๋ณ๋ฆฌํ๊ณผ/์์ฌ๊ฒฐํต๊ท ์ ๊ฒ์ถ์ ์ํด ์ฌ์ฉ๋๋ ์ธ๊ท ํ์ ๊ฒ์ฌ๋ฒ์ ์ค๋ ์๊ฐ์ด ์์๋๊ณ ๋ฏผ๊ฐ๋์ ํน์ด๋์ ํ๊ณ๊ฐ ์์ด ์ต๊ทผ PCR์ ์ด์ฉํ ํต์ฐ์ฆํญ๋ฒ์ด ๊ฒฐํต๊ฒ์ฌ์ ๋ง์ด ์ด์ฉ๋๊ณ ์๋ค. ๊ทธ๋ฌ๋ ํ์ฌ๊น์ง ์ฌ์ฉ๋๊ณ ์๋ ๊ฒฐํต๊ฒ์ฌ์ฉ ํต์ฐ์ฆํญ๋ฒ์ ๋๋ถ๋ถ DNA๋ฅผ ํ์ ์ผ๋ก ํ๊ณ ์๋ค. ๊ทธ๋ฐ๋ฐ ๊ฐ์ ์์ ์ธํฌ์ผ ๊ฒฝ์ฐ ํน์ ์ ์ ์์ DNA๋ณด๋ค RNA๊ฐ ๋ ๋ง์ ์๋ก ์กด์ฌํ๋ค. ๋ฐ๋ผ์ RNA๋ฅผ ํ์ ์ผ๋ก ํ๋ ํต์ฐ์ฆํญ๋ฒ์ ๋ฏผ๊ฐ๋๊ฐ ๋ ๋์ ์ ์๋ค. ๋ํ DNA ๋ณต์ ๊ณผ์ ์ ์์ฉํ์ฌ ๊ฐ๋ฐ๋ PCR๋ณด๋ค ์ ์ฌ๋ RNA๋ก๋ถํฐ DNA๋ฅผ ํฉ์ฑํ๊ณ ํฉ์ฑ๋ DNA๋ก๋ถํฐ ๋ง์ ์์ RNA๋ฅผ ๋ค์ ์ ์ฌํ๋ nucleic acid sequence-based amplification (NASBA)์ ์ด์ฉํ๋ค๋ฉด ๊ฒฐํต๊ฒ์ฌ์ ๋ฏผ๊ฐ๋๋ฅผ ๋ ๋์ผ ์ ์์ ๊ฒ์ด๋ค. ์ด์ ๋ณธ ์ฐ๊ตฌ์์๋ Mycobacterium bovis BCG์์ ์ถ์ถํ DNA์ RNA๋ฅผ ์ด์ฉํ์ฌ PCR, RT-PCR, real-time RT-PCR, real-time NASBA๋ฅผ ์ํํ์ฌ DNA ๋ฐ RNA๋ฅผ ํ์ ์ผ๋ก ํ ๊ฒฐํต๊ฒ์ฌ๋ฒ์ ๋ฏผ๊ฐ๋๋ฅผ ๋น๊ตํ๋ ํํธ, PCR ๋ฐ NASBA๋ฅผ ์ด์ฉํ ๊ฒฐํต๊ฒ์ฌ๋ฒ์ ๋ฏผ๊ฐ๋๋ฅผ ๋น๊ตํ์๋ค. ๊ทธ ๊ฒฐ๊ณผ, 16S rRNA ์ ์ ์๋ฅผ ํ์ ์ผ๋ก ํ PCR์ ๋ฏผ๊ฐ๋๋ 10 3 cfu/ใ ๋ฐ 100 pg์ด์๊ณ , 16S rRNA๋ฅผ ํ์ ์ผ๋ก ํ RT-PCR์ ๋ฏผ๊ฐ๋๋ 10 2 cfu/ใ ๋ฐ 10 pg๋ก ๋ํ๋ DNA๋ณด๋ค RNA๋ฅผ ํ์ ์ผ๋ก ํ ๊ฒฐํต๊ฒ์ฌ๋ฒ์ด ๋ ๋ฏผ๊ฐํ์๋ค. ๋ํ rpoB mRNA์ 16S rRNA๋ฅผ ๊ฐ๊ฐ ํ์ ์ผ๋ก ํ์ฌ RT-PCR์ ์ํํ ๊ฒฐ๊ณผ, rpoB mRNA๋ฅผ ํ์ ์ผ๋ก ํ์์ ๋์ ๋ฏผ๊ฐ๋๊ฐ 10 5 cfu/ใ์ธ ๋ฐ๋ฉด 16S rRNA๋ฅผ ํ์ ์ผ๋ก ํ RT-PCR์ ๋ฏผ๊ฐ๋๊ฐ 10 2 cfu/ใ๋ก ๋ํ๋ RNA์ ํ์ ์ ๋ฐ๋ผ ๋ฏผ๊ฐ๋๊ฐ ๋ฌ๋ผ์ง์ ์ ์ ์์๋ค. ์ด์ด 16S rRNA๋ฅผ ํ์ ์ผ๋ก ํ RT-PCR๊ณผ ํ๊ด ํ์ง probe๋ฅผ ์ด์ฉํ real-time RT-PCR ๋ฐ real-time NASBA๋ฅผ ์ํํ์ฌ ํต์ฐ์ฆํญ๋ฒ์ ๋ฐ๋ฅธ ๋ฏผ๊ฐ๋๋ฅผ ๋น๊ตํ ๊ฒฐ๊ณผ, real-time RT-PCR์์ 10 fg๊ณผ real-time NASBA 1 fg์ ๋ฏผ๊ฐ๋๋ฅผ ํ์ธํ์๋ค. ๋ฐ๋ผ์ RNA๋ฅผ ํ์ ์ผ๋ก real-time RT-PCR, real- time NASBA ๋ฑ์ ์ด์ฉํ์ฌ ๊ฒฐํต๊ท ์ ๊ฒ์ถํ ๊ฒฝ์ฐ ๋ณด๋ค ๋ฏผ๊ฐํ ๊ฒฐํต๊ฒ์ฌ๊ฐ ๊ฐ๋ฅํ ๊ฒ์ผ๋ก ์ฌ๋ฃ๋๋ค.ope
๋ง์ฑ ๋น๋ถ๋น๋์ผ์ด ๋๋ฐ๋ ๋น์ฉ์์์ ํธ์ฐ๊ตฌ ์นจ์ค ์ ๋์ ๋ฐ๋ฅธ ์กฐ์งํ์ ํน์ฑ
ํ์๋
ผ๋ฌธ(์์ฌ)--์์ธ๋ํ๊ต ๋ํ์ :์ํ๊ณผ ์ด๋น์ธํ๊ณผํ์ ๊ณต,2006.Maste
ํธ์ฐ๊ตฌ์ฑ ๋น๋ถ๋น๋์ผ์ ๋น์ฉ ํ์ฑ์์ interleukin-17A์ ์ญํ
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ผ๋ฌธ (๋ฐ์ฌ)-- ์์ธ๋ํ๊ต ๋ํ์ : ์ํ๊ณผ ๋ฉด์ญํ์ ๊ณต, 2015. 8. ์ด์ฌ์.Background: Nasal polyposis associated with chronic rhinosinusitis (CRS) is a chronic inflammatory disease which is characterized by infiltration of many inflammatory cells. Meanwhile, interleukin (IL)-17A is a well-known proinflammatory cytokine which induces both eosinophilic and neutrophilic inflammation. We investigated the role of IL-17A in the development of nasal polyps in the CRS murine model.
Materials & Methods: Eosinophilic CRS with nasal polyps was induced by using ovalbumin (OVA) and Staphylococcus aureus enterotoxin B (SEB) in wild-type BALB/c and IL-17A knock-out (KO) mice. Histopathologic changes of the sinonasal cavity were evaluated using hematoxylin and eosin, Periodic acid-Schiff, Sirius red, Massonโs trichrome and immunohistochemistry. The levels of cytokines in splenocyte supernatants and total and OVA-specific IgEs in sera were measured using enzyme-linked immunosorbent assay. The expression levels of cytokines in the nasal mucosa were assessed by quantitative real-time polymerase chain reaction.
Results: Under the IL-17A deficiency, cytokines (IL-4, IL-5 and interferon-ฮณ) in splenocyte supernatants as well as total and OVA-specific IgEs in sera were reduced significantly. Infiltration of both eosinophils and neutrophils into the nasal mucosa, subepithelial fibrosis and goblet cell count also decreased significantly in IL-17A KO mice treated with both OVA and SEB compared with those in the wild-type counterpart. However, there were no significant differences in either numbers or sizes of polypoid lesions among groups. Meanwhile, IL-4 increased and interferon-ฮณ decreased in the nasal mucosa in IL-17A KO mice treated with both OVA and SEB.
Conclusions: This study suggests that even though IL-17A plays an important role in both local inflammation and remodeling of the nasal mucosa in CRS, it cannot entirely account for the development of nasal polyps.Abstract ----------------------------------------------- i
Contents --------------------------------------------- iv
List of Tables ----------------------------------------- v
List of Figures ---------------------------------------- vi
List of Abbreviations --------------------------------- viii
Introduction ------------------------------------------- 1
Materials and Methods -------------------------------- 3
Results ---------------------------------------------- 10
Discussion ------------------------------------------ 14
References ------------------------------------------ 21
Tables ---------------------------------------------- 28
Figures ---------------------------------------------- 29
Abstract (Korean) ------------------------------------ 40Docto