PMID: 11337047May 5, 2001Paper

Comparison of different methods of total RNA extraction for viral detection in sputum

Journal of Virological Methods
X XiangW C Tan

Abstract

Examination of sputum specimens can be used for monitoring airway inflammation by means of immunological and molecular techniques. RNA extraction is essential for measurement of cytokine gene expression and for detection of many viral pathogens in sputum. In this study, three RNA extraction methods used commonly (acid guanidinium thiocyanate-phenol-chloroform extraction, Trizol and RNeasy Mini kit) were compared on the sputum of 14 patients who had chronic obstructive pulmonary disease. The effect of dithiothreitol pre-treatment on sputum RNA extraction was also investigated. The yield and purity of total RNA were determined by spectrophotometry. Reverse transcription-polymerase chain reaction (RT-PCR) results of the house keeping gene (glyceraldehyde-3-phosphate dehydrogenase) and RNA sequences specific to common respiratory viruses were compared. The results showed that (1) total RNA extracted with Trizol had highest yield and purity among the three RNA extraction methods; (2) there was no significant difference among the three RNA extraction methods on the house keeping gene and viral detection by RT-PCR; (3) dithiothreitol pre-treatment did not improve either the purity of total RNA, or RT-PCR signal. Moreover, dithiothreit...Continue Reading

References

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Citations

Oct 22, 2008·Applied Biochemistry and Biotechnology·Dongmin LiShemin Lu
Nov 6, 2013·Biosecurity and Bioterrorism : Biodefense Strategy, Practice, and Science·Simona Di PasqualeLivia Di Trani
May 11, 2004·Annals of Clinical Microbiology and Antimicrobials·James M McMahonMichael Clatts
Feb 11, 2010·Future Microbiology·Patrick Tang, Charles Chiu
Aug 19, 2014·Journal of Clinical Virology : the Official Publication of the Pan American Society for Clinical Virology·M J VanspauwenC F M Linssen
May 7, 2014·Journal of Medical Virology·Ji Hun JeongYiel Hea Seo
Sep 6, 2005·Analytical Biochemistry·Stacey L HembruffAmadeo M Parissenti
Sep 15, 2006·Analytical Biochemistry·David A MacIntyreEng-Cheng Chan
Apr 1, 2015·Biopreservation and Biobanking·Yaejee H HongRobert B Hinton
Aug 19, 2014·Analytical Biochemistry·Juliana T Y LeeVictor Y L Leung
Feb 16, 2006·Journal of Veterinary Diagnostic Investigation : Official Publication of the American Association of Veterinary Laboratory Diagnosticians, Inc·Ming Y DengThomas S McKenna
Jul 3, 2015·International Journal of Environmental Research and Public Health·Stefan PanaiotovTodor Kantardjiev
Jun 29, 2004·World Journal of Gastroenterology : WJG·Song-Ming GuoWei Yang
Dec 4, 2008·Critical Reviews in Biotechnology·Jai Prakash MuyalHeinz Fehrenbach
Nov 2, 2017·American Journal of Respiratory Cell and Molecular Biology·Csilla PaskaBalazs Antus
Aug 15, 2019·Analytical Chemistry·Duane S JuangDavid J Beebe

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