Comparison of the Effectiveness in the Application of Competitive and Noncompetitive Internal Control for the Laboratory Developed Polymerase Chain Reaction
10.17945/kjbt.2019.30.1.57
- Author:
Sunmi SHIN
1
;
Jung Won KANG
;
Jae won KANG
;
Young Ik SEO
;
Hyukki MIN
Author Information
1. Blood Transfusion Research Institute, Korean Red Cross, Wonju, Korea. kangjaewon@redcross.or.kr
- Publication Type:Original Article
- Keywords:
Human T-cell lymphotropic virus;
Laboratory-developed PCR;
Competitive internal control;
Noncompetitive internal control
- MeSH:
DNA;
Genes, Essential;
Glyceraldehyde 3-Phosphate;
Limit of Detection;
Nucleic Acid Amplification Techniques;
Oxidoreductases;
Plasmids;
Polymerase Chain Reaction
- From:Korean Journal of Blood Transfusion
2019;30(1):57-64
- CountryRepublic of Korea
- Language:Korean
-
Abstract:
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.
