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http://dx.doi.org/10.15324/kjcls.2019.51.3.309

Evaluation of Commercial Complementary DNA Synthesis Kits for Detecting Human Papillomavirus  

Yu, Kwangmin (Department of Biomedical Laboratory Science, College of Health Sciences, Yonsei University)
Park, Sunyoung (Department of Mechanical Engineering, Yonsei University)
Chang, Yunhee (Department of Biomedical Laboratory Science, College of Health Sciences, Yonsei University)
Hwang, Dasom (Department of Biomedical Laboratory Science, College of Health Sciences, Yonsei University)
Kim, Geehyuk (Ministry of Food and Drug Safety Pharmaceutical Safety Bureau, Osong Health Technology Administration Complex)
Kim, Jungho (Clinical Vaccine Research Section, International Tuberculosis Research Center)
Kim, Sunghyun (Department of Clinical Laboratory Science, College of Health Sciences, Catholic University of Pusan)
Kim, Eun-Joong (Department of Clinical Laboratory Science, Chungbuk Health and Science University)
Lee, Dongsup (Department of Clinical Laboratory Science, Hyejeon College)
Publication Information
Korean Journal of Clinical Laboratory Science / v.51, no.3, 2019 , pp. 309-315 More about this Journal
Abstract
Cervical cancer is the fourth most common malignant neoplasm in women worldwide. Most cases of cervical cancer are caused by an infection by the human papillomavirus. Molecular diagnostic methods have emerged to detect the HPV for sensitivity, specificity, and objectivity. In particular, real-time PCR has been introduced to acquire a more sensitive target DNA or RNA. RNA extraction and complementary DNA synthesis are proceeded before performing real-time PCR targeting RNA. To identify an adequate and sensitive cDNA synthesis kit, this study evaluated the two commonly used kits for cDNA synthesis. The results show that the $R^2$ and efficiency (%) of the two cDNA synthesis kits were similar in the cervical cancer cell lines. On the other hand, the Takara kit compared to Invitrogen kit showed P<0.001 in the $10^2$ and $10^3$ SiHa cell count. The Takara kit compared to the Invitrogen kit showed P<0.001 in the $10^1$ and $10^2$ HeLa cell count. Furthermore, 8, 4, 2, 1, and 0.5 ml of forty exfoliated cell samples were used to compare the cDNA synthesis kits. The Takara kit compared to the Invitrogen kit showed P<0.01 in 8, 4, and 1 ml and P<0.05 in 0.5 mL. The study was performed to identify the most appropriate cDNA synthesis kit and suggests that a cDNA synthesis kit could affect the real-time PCR results.
Keywords
Cervical cancer; Complementary DNA; Complementary DNA synthesis kit; Human papillomavirus; Real-time PCR;
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