Detection of viral infections by in situ PCR: theoretical considerations and possible value in diagnostic pathology

Journal of Clinical Laboratory Analysis
G J Nuovo

Abstract

The technique that allows for in situ detection of PCR-amplified DNA and cDNA has improved remarkably quickly over the past 5 years. As of this writing, one can expect good results after a relatively quick "learning curve." The technique demands some skill in working with glass slides and cells/tissues. Thus, I recommend that one without experience in tissue analysis spend a few weeks working with either standard in situ hybridization or immunohistochemistry to be comfortable in that regard. Commercially available kits for in situ hybridization are good and readily available (e.g., Enzo Diagnostics, NY). When ready to begin in situ PCR, the most user-friendly system now available is from Perkin Elmer. I cannot overstate the importance of performing the needed controls with each and every run, of the crucial relationship between protease digestion time and success with RT in situ PCR, and the need to use relatively high (4.5 mM) MgCl2 and to include bovine serum albumin (or increased Taq polymerase). Given these improvements and the important information available from in situ PCR that cannot be provided by standard in situ hybridization or solution-phase PCR, one would anticipate that in situ PCR will someday enter the field of...Continue Reading

References

Nov 1, 1992·PCR Methods and Applications·G J NuovoP D Gorevic
Mar 15, 1992·Proceedings of the National Academy of Sciences of the United States of America·F NegroF Bonino
Mar 1, 1992·The Journal of Histochemistry and Cytochemistry : Official Journal of the Histochemistry Society·K P ChiuG W Jordan
Jul 1, 1990·Proceedings of the National Academy of Sciences of the United States of America·A T HaaseK A Staskus
Feb 1, 1986·Proceedings of the National Academy of Sciences of the United States of America·M E HarperF Wong-Staal
Jun 1, 1995·Diagnostic Molecular Pathology : the American Journal of Surgical Pathology, Part B·K LidonniciG J Nuovo
Oct 1, 1994·PCR Methods and Applications·G J NuovoF Gallery
Jul 1, 1993·The American Journal of Surgical Pathology·G J NuovoB Lane
May 1, 1993·PCR Methods and Applications·G J NuovoW Bloch

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Citations

May 8, 2001·Clinical Infectious Diseases : an Official Publication of the Infectious Diseases Society of America·G W Procop, M Wilson
Jan 20, 2010·Archives of Medical Research·Shih-Chi LiuChuen-Den Tseng
Aug 15, 1998·Biochemical and Biophysical Research Communications·N NakajimaH Yoshikura
Jan 20, 2005·Journal of Biochemical and Biophysical Methods·Changbaig HyunIan Hughes
Oct 24, 1998·Diagnostic Molecular Pathology : the American Journal of Surgical Pathology, Part B·T B PlummerR V Lloyd
Apr 23, 2009·Indian Journal of Medical Microbiology·E GuptaT Singh

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