Background: Extraction of viral nucleic acids from serum samples is widely used in diagnostic pathology tests. However, the heterogeneous nature of non-serum samples may contribute to variations in the yields of viral nucleic acids with different extraction methods and specimen types.
Objectives: Six different methods were compared for optimal extraction of viral DNA or RNA from four types of non-serum specimens.
Study design: The DNA viruses used were herpes simplex virus and cytomegalovirus. The RNA viruses were poliovirus, rotavirus and small round structured virus. The specimens used were from respiratory, genital, faecal and peripheral blood mononuclear cell samples. The extracted nucleic acids were amplified by PCR and detected in an enzyme immunoassay using digoxygenin-labelled amplicons.
Results and conclusions: For extraction of viral DNA, the phenol-chloroform method yielded the highest amount of DNA as judged by endpoint titration. The three methods compared for extraction of viral RNA used guanidine isothiocyanate and the QiaRNA kit was shown to yield the highest amount of viral RNA.
Rasool NB, Monroe SS, Glass RI. Rasool NB, et al. J Virol Methods. 2002 Feb;100(1-2):1-16. doi: 10.1016/s0166-0934(01)00379-2. J Virol Methods. 2002. PMID: 11742648
Akhtar N, Ni J, Stromberg D, Rosenthal GL, Bowles NE, Towbin JA. Akhtar N, et al. Circulation. 1999 Apr 20;99(15):2011-8. doi: 10.1161/01.cir.99.15.2011. Circulation. 1999. PMID: 10209006
Espy MJ, Rys PN, Wold AD, Uhl JR, Sloan LM, Jenkins GD, Ilstrup DM, Cockerill FR 3rd, Patel R, Rosenblatt JE, Smith TF. Espy MJ, et al. J Clin Microbiol. 2001 Jun;39(6):2233-6. doi: 10.1128/JCM.39.6.2233-2236.2001. J Clin Microbiol. 2001. PMID: 11376062 Free PMC article.
Hale AD, Green J, Brown DW. Hale AD, et al. J Virol Methods. 1996 Apr 5;57(2):195-201. doi: 10.1016/0166-0934(95)01966-9. J Virol Methods. 1996. PMID: 8801231
McDougall JK, Nelson JA, Myerson D, Beckmann AM, Galloway DA. McDougall JK, et al. J Invest Dermatol. 1984 Jul;83(1 Suppl):72s-76s. doi: 10.1111/1523-1747.ep12281204. J Invest Dermatol. 1984. PMID: 6330227 Review.
Kiyasu Y, Akashi Y, Sugiyama A, Takeuchi Y, Notake S, Naito A, Nakamura K, Ishikawa H, Suzuki H. Kiyasu Y, et al. Mol Diagn Ther. 2021 Jul;25(4):495-504. doi: 10.1007/s40291-021-00535-5. Epub 2021 May 18. Mol Diagn Ther. 2021. PMID: 34003471 Free PMC article.
Esona MD, McDonald S, Kamili S, Kerin T, Gautam R, Bowen MD. Esona MD, et al. J Virol Methods. 2013 Dec;194(1-2):242-9. doi: 10.1016/j.jviromet.2013.08.023. Epub 2013 Sep 13. J Virol Methods. 2013. PMID: 24036075 Free PMC article.
Miao L, Yin RX, Hu XJ, Wu DF, Cao XL, Li Q, Yan TT, Aung LH, Wu JZ, Lin WX. Miao L, et al. Lipids Health Dis. 2012 May 30;11:61. doi: 10.1186/1476-511X-11-61. Lipids Health Dis. 2012. PMID: 22646906 Free PMC article.
Garcia-Sepulveda CA, Carrillo-Acuña E, Guerra-Palomares SE, Barriga-Moreno M. Garcia-Sepulveda CA, et al. Mol Biol Rep. 2010 Apr;37(4):1883-90. doi: 10.1007/s11033-009-9624-1. Epub 2009 Jul 17. Mol Biol Rep. 2010. PMID: 19609718
Lu YQ, Han JX, Qi P, Xu W, Zu YH, Zhu B. Lu YQ, et al. World J Gastroenterol. 2006 Dec 7;12(45):7365-70. doi: 10.3748/wjg.v12.i45.7365. World J Gastroenterol. 2006. PMID: 17143958 Free PMC article.