DNA extraction from paraffin embedded colorectal carcinoma samples: A comparison study of manual vs automated methods,using four commercially kits

作     者：Zsolt Kovacs Ioan Jung Erzsebet Csernak Zoltan Szentirmay Laura Banias Genoveva Rigmanyi Simona Gurzu 

作者机构：Department of PathologyUniversity of MedicinePharmacySciences and TechnologyTargu-Mures 530149Romania Research Center(CCAMF)University of MedicinePharmacySciences and TechnologyTargu-Mures 540139Romania 

出 版 物：《World Journal of Clinical Oncology》 (世界临床肿瘤学杂志（英文版）)

年 卷 期：2019年第10卷第9期

页      面：307-317页

学科分类：1002[医学-临床医学] 100214[医学-肿瘤学] 10[医学] 

基　　金：the University of Medicine Pharmacy Science and Technology Research Grant No.275/11.01.2017 

主　　题：DNA isolation Colorectal cancer Paraffin-embedded PureLink Genomic DNA Mini Kit High Pure FFPE DNA Isolation Kit iPrep Genomic DNA Kit MagnaPure LC DNA Isolation Kit 

摘      要：BACKGROUND Nucleic acid isolation from formalin-fixed, paraffin-embedded tissue(FFPET)samples is a daily routine in molecular pathology laboratories, but extraction from FFPET is not always easily achieved. Choosing the right extraction technique is key for further *** To compare the performance of four commercially available kits used for DNA extraction in routine *** DNA isolation was performed on 46 randomly selected formalin-fixed, paraffinembedded(FFPE) colorectal adenocarcinoma(CRC) surgical specimens. Four commercially available extraction kits were used: two for manual DNA extraction(the Pure Link Genomic DNA Mini Kit from Invitrogen and the High Pure FFPE DNA Isolation Kit from Roche) and two for automated DNA extraction(the i Prep Genomic DNA Kit from Invitrogen and the Magna Pure LC DNA Isolation Kit from Roche). The DNA concentration and quality(odds ratio) among the four systems were compared. The results were correlated with the clinicopathological aspects of CRC cases: age, gender, localization, macro-and microscopic features,lymph node metastases, and the lymph node *** The highest DNA concentration was obtained using the manual kits: 157.24 ±62.99 ng/μL for the Pure Link Genomic DNA Mini Kit and 86.64 ng/μL± 43.84 for the High Pure FFPE DNA Isolation Kit(P 0.0001). Lower concentrations were obtained with automated systems: 20.39 ± 21.19 ng/μL for the Magna Pure LC DNA Isolation Kit and 8.722 ± 6.408 ng/μL for the i Prep Genomic DNA Kit,with differences between the systems used(P 0.0001). The comparison between age, gender, tumor localization, pT or pN stage and the lymph node ratio indicated no statistically significant difference in DNA concentration using any of the nucleic acid isolation kits. DNA concentration was influenced by the macroscopic features and grade of differentiation. A higher DNA concentration was obtained for well-differentiated polypoid colorectal adenocarcinomas(CRCs), compared with undif