Affiliations: [a] Department of Pediatrics, Kyungpook National University School of Medicine, Daegu, Korea | [b] Department of Physiology, Kyungpook National University School of Medicine, Daegu, Korea | [c] Department of Emergency Medicine, Kyungpook National University School of Medicine, Daegu, Korea
Correspondence:
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Corresponding author: Jong Kun Kim, Department of Emergency Medicine, Kyungpook National University School of
Abstract: BACKGROUND: 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. OBJECTIVE: 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. METHODS: 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. RESULTS: 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. CONCLUSIONS: 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.
