Comparison of methods for extraction of viral DNA from cellular specimens

被引:6
作者
Cook, Linda [2 ]
Atienza, Ederlyn E. [2 ]
Bagabag, Arthur [2 ]
Obrigewitch, Rose M. [2 ]
Jerome, Keith R. [1 ,2 ]
机构
[1] Fred Hutchinson Canc Res Ctr, Vaccine & Infect Dis Inst, Seattle, WA 98109 USA
[2] Univ Washington, Ctr Med, Dept Lab Med, Seattle, WA 98195 USA
关键词
PCR; Nucleic acid; Extraction; Tissue; NUCLEIC-ACID EXTRACTION; REAL-TIME PCR; EPSTEIN-BARR-VIRUS; TRANSPLANT RECIPIENTS; CLINICAL SPECIMENS; NUCLISENS EASYMAG; SYSTEMS; BLOOD; RNA;
D O I
10.1016/j.diagmicrobio.2009.01.003
中图分类号
R51 [传染病];
学科分类号
100401 ;
摘要
The accuracy and precision of quantitative polymerase chain reaction (PCR) results depend not only on the PCR reaction but also on the extraction of viral nucleic acid. Although the extraction of viral nucleic acid from fluids has been extensively studied, less data are available regarding extractions from cellular specimens. We therefore evaluated several commercially available kits for the extraction of nucleic acid from cellular specimens. Although the kits generally performed well, there were some differences in extraction efficiency, especially at low numbers of input cells. Inclusion of a multivirus positive control allowed careful monitoring of extraction efficiency during routine clinical use. Laboratories are encouraged to validate extraction methods carefully in the context of the proposed viral testing. (C) 2009 Elsevier Inc. All rights reserved.
引用
收藏
页码:37 / 42
页数:6
相关论文
共 13 条
[1]   Automated extraction of viral-pathogen RNA and DNA for high-throughput quantitative real-time PCR [J].
Beuselinck, K ;
van Ranst, M ;
van Eldere, J .
JOURNAL OF CLINICAL MICROBIOLOGY, 2005, 43 (11) :5541-5546
[2]   Optimization of quantitative detection of cytomegalovirus DNA in plasma by real-time PCR [J].
Boeckh, M ;
Huang, M ;
Ferrenberg, J ;
Stevens-Ayers, T ;
Stensland, L ;
Nichols, WG ;
Corey, L .
JOURNAL OF CLINICAL MICROBIOLOGY, 2004, 42 (03) :1142-1148
[3]   Comparison of the NucliSens easyMAG and Qiagen BioRobot 9604 nucleic acid extraction systems for detection of RNA and DNA respiratory viruses in nasopharyngeal aspirate samples [J].
Chan, Kwok Hung ;
Yam, Wing Cheong ;
Pang, Chiu Mei ;
Chan, Kit Man ;
Lam, Siu Yan ;
Lo, Kam Fai ;
Poon, Leo L. M. ;
Peiris, J. S. Malik .
JOURNAL OF CLINICAL MICROBIOLOGY, 2008, 46 (07) :2195-2199
[4]   Comparison of automated nucleic acid extraction methods with manual extraction [J].
Dundas, Nicola ;
Leos, N. Kristine ;
Mitui, Midori ;
Revell, Paula ;
Rogers, Beverly Barton .
JOURNAL OF MOLECULAR DIAGNOSTICS, 2008, 10 (04) :311-316
[5]  
Gerna Giuseppe, 2006, Herpes, V13, P4
[6]   Quantitative stability of DNA after extended storage of clinical specimens as determined by real-time PCR [J].
Jerome, KR ;
Huang, ML ;
Wald, A ;
Selke, S ;
Corey, L .
JOURNAL OF CLINICAL MICROBIOLOGY, 2002, 40 (07) :2609-2611
[7]   Markers of viral infection in monozygotic twins discordant for chronic fatigue syndrome [J].
Koelle, DM ;
Barcy, S ;
Huang, ML ;
Ashley, RL ;
Corey, L ;
Zeh, J ;
Ashton, S ;
Buchwald, D .
CLINICAL INFECTIOUS DISEASES, 2002, 35 (05) :518-525
[8]   Detection of Epstein-Barr virus DNA in sera from transplant recipients with lymphoproliferative disorders [J].
Limaye, AP ;
Huang, ML ;
Atienza, EE ;
Ferrenberg, JM ;
Corey, L .
JOURNAL OF CLINICAL MICROBIOLOGY, 1999, 37 (04) :1113-1116
[9]   Quantitation of BK virus load in serum for the diagnosis of BK virus-associated nephropathy in renal transplant recipients [J].
Limaye, AP ;
Jerome, KR ;
Kuhr, CS ;
Ferrenberg, J ;
Huang, ML ;
Davis, CL ;
Corey, L ;
Marsh, CL .
JOURNAL OF INFECTIOUS DISEASES, 2001, 183 (11) :1669-1672
[10]   Evaluation of NucliSens easyMAG for automated nucleic acid extraction from various clinical specimens [J].
Loens, K. ;
Bergs, K. ;
Ursi, D. ;
Goossens, H. ;
Ieven, M. .
JOURNAL OF CLINICAL MICROBIOLOGY, 2007, 45 (02) :421-425