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- Title
A comparative study of three different gene expression analysis methods.
- Authors
Jae Young Choe; Hyung Soo Han; Seon Duk Lee; Hanna Lee; Dong Eun Lee; Jae Yun Ahn; Hyun Wook Ryoo; Kang Suk Seo; Jong Kun Kim; Choe, Jae Young; Han, Hyung Soo; Lee, Seon Duk; Lee, Hanna; Lee, Dong Eun; Ahn, Jae Yun; Ryoo, Hyun Wook; Kim, Jong Kun; Seo, Kang Suk
- Abstract
<bold>Background: </bold>TNF-α regulates immune cells and acts as an endogenous pyrogen. Reverse transcription polymerase chain reaction (RT-PCR) is one of the most commonly used methods for gene expression analysis. Among the alternatives to PCR, loop-mediated isothermal amplification (LAMP) shows good potential in terms of specificity and sensitivity.<bold>Objective: </bold>However, few studies have compared RT-PCR and LAMP for human gene expression analysis. Therefore, in the present study, we compared one-step RT-PCR, two-step RT-LAMP and one-step RT-LAMP for human gene expression analysis.<bold>Methods: </bold>We compared three gene expression analysis methods using the human TNF-α gene as a biomarker from peripheral blood cells. Total RNA from the three selected febrile patients were subjected to the three different methods of gene expression analysis.<bold>Results: </bold>In the comparison of three gene expression analysis methods, the detection limit of both one-step RT-PCR and one-step RT-LAMP were the same, while that of two-step RT-LAMP was inferior. One-step RT-LAMP takes less time, and the experimental result is easy to determine.<bold>Conclusions: </bold>One-step RT-LAMP is a potentially useful and complementary tool that is fast and reasonably sensitive. In addition, one-step RT-LAMP could be useful in environments lacking specialized equipment or expertise.
- Subjects
GENE expression; TUMOR necrosis factors; REVERSE transcriptase polymerase chain reaction; BLOOD cells; PYROGENS; COMPARATIVE studies; DNA probes; RESEARCH methodology; MEDICAL cooperation; POLYMERASE chain reaction; RESEARCH; EVALUATION research; GENE expression profiling; NUCLEIC acid amplification techniques; STANDARDS
- Publication
Technology & Health Care, 2017, Vol 25, Issue 6, p1073
- ISSN
0928-7329
- Publication type
journal article
- DOI
10.3233/THC-170967