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Evaluation of Commercial Complementary DNA Synthesis Kits for Detecting Human Papillomavirus
Korean J Clin Lab Sci 2019;51:309-315  
Published on September 30, 2019
Copyright © 2019 Korean Society for Clinical Laboratory Science.

Kwangmin Yu1, Sunyoung Park2, Yunhee Chang1, Dasom Hwang1, Geehyuk Kim3, Jungho Kim4, Sunghyun Kim5, Eun-Joong Kim6, Dongsup Lee7

1Department of Biomedical Laboratory Science, College of Health Sciences, Yonsei University, Wonju, Korea
2Department of Mechanical Engineering, Yonsei University, Seoul, Korea
3Ministry of Food and Drug Safety Pharmaceutical Safety Bureau, Osong Health Technology Administration Complex, Osong, Korea
4Clinical Vaccine Research Section, International Tuberculosis Research Center, Seoul, Korea
5Department of Clinical Laboratory Science, College of Health Sciences, Catholic University of Pusan, Pusan, Korea
6Department of Clinical Laboratory Science, Chungbuk Health and Science University, Cheongju, Korea
7Department of Clinical Laboratory Science, Hyejeon College, Hongseong, Korea
Correspondence to: * Dongsup Lee
Department of Clinical Laboratory Science, Hyejeon College, 25 Daehak-gil, Hongseong 32244, Korea
E-mail: eastern3547@naver.com
* ORCID: https://orcid.org/0000-0003-4375-2731
This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/4.0) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.
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 R2 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 102 and 103 SiHa cell count. The Takara kit compared to the Invitrogen kit showed P<0.001 in the 101 and 102 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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  • Hyejeon College