지원사업
학술연구/단체지원/교육 등 연구자 활동을 지속하도록 DBpia가 지원하고 있어요.
커뮤니티
연구자들이 자신의 연구와 전문성을 널리 알리고, 새로운 협력의 기회를 만들 수 있는 네트워킹 공간이에요.
초록· 키워드
오류제보하기
Background: A nucleic acid amplification test was adopted to detect transfusion-transmitted infectious agents. In the case of HTLV, however, there was no internal control (IC) because the laboratory developed polymerase chain reaction (laboratory-developed PCR) was used. In this study, noncompetitive IC was constructed for the laboratory-developed PCR of HTLV and the effectiveness was compared with the competitive test that was constructed in a previous study. Methods: As a competitive IC, plasmid DNA, including the primer recognition sequence for the amplification of the HTLV pX region, was constructed. As a noncompetitive IC, an additional primer was constructed for the amplification of the housekeeping gene, the glyceraldehyde 3-phosphate dehydrogenase (GAPDH) gene. The performance of the competitive and noncompetitive IC was verified and compared using 10 HTLV positive samples and 10 negative samples. In addition, the detection limits in the assay adopting competitive IC and noncompetitive IC were compared. Results: In the case of competitive IC applications, all 10 positive samples were positive and all 10 negative samples were negative. In the case of noncompetitive IC applications, however, one positive sample was not detected. The detection limit of the assay using competitive IC was 100 pg and that of the assay using noncompetitive IC was 1 ng. Conclusion: Although the manufacturing processes is not required using noncompetitive IC, the adoption of competitive IC is more effective to ensure the assay results because the ability of detection of the assay adopting competitive IC was better than that using noncompetitive IC.
목차
등록된 정보가 없습니다.
참고문헌 (13)
참고문헌 신청
Hoorfar J / 2004 / Practical considerations in design of internal amplification controls for diagnostic PCR assays / J Clin Microbiol 42:1863~1868
Murphy NM / 2007 / Construction and evaluation of a microbiological positive process internal control for PCR-based examination of food samples for Listeria monocytogenes and Salmonella enterica / Int J Food Microbiol 120:110~119
Sohni Y / 2008 / Cloning and development of synthetic internal amplification control for Bacillus anthracis real-time polymerase chain reaction assays / Diagn Microbiol Infect Dis 61:471~475
강정원 / 2018 / 실험실에서 자체 설계한 HTLV PCR에서의 내부정도관리물질 제조 및 적용 / 대한수혈학회지 29(1):33~40
Bas A / 2004 / Utility of the housekeeping genes 18S rRNA, beta-actin and glyceraldehyde-3-phosphate-dehydrogenase for normalization in real-time quantitative reverse transcriptasepolymerase chain reaction analysis of gene expression in human T lymphocytes / Scand J Immunol 59:566~573
Murphy NM / 2007 / Construction and evaluation of a microbiological positive process internal control for PCR-based examination of food samples for Listeria monocytogenes and Salmonella enterica / Int J Food Microbiol 120:110~119
Sohni Y / 2008 / Cloning and development of synthetic internal amplification control for Bacillus anthracis real-time polymerase chain reaction assays / Diagn Microbiol Infect Dis 61:471~475
강정원 / 2018 / 실험실에서 자체 설계한 HTLV PCR에서의 내부정도관리물질 제조 및 적용 / 대한수혈학회지 29(1):33~40
Bas A / 2004 / Utility of the housekeeping genes 18S rRNA, beta-actin and glyceraldehyde-3-phosphate-dehydrogenase for normalization in real-time quantitative reverse transcriptasepolymerase chain reaction analysis of gene expression in human T lymphocytes / Scand J Immunol 59:566~573
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