Abstract
The ability to detect mycobacterial DNA by polymerase chain reaction (PCR)-based methodology in formalin-fixed, paraffin-embedded tissue sections is useful in several clinical scenarios. The major use of this type of assay is in those instances in which infectious disease is not clinically suspected and microbiological cultures are not performed. In these cases, the only tissue available for examination is that present in routinely prepared paraffin blocks after histologic examination. The presence of necrotizing granulomatous inflammation should result in special stains for acid-fast organisms. However, in many such cases the special stains are unsatisfactory, because the number of organisms present is very low (as in Mycobacterium tuberculosis). Thus, the tedious examination of multiple serial sections is often necessary to identify the pathogenic organism, and often no organism is found. Therefore, more sensitive detection methods are needed. PCR-based detection of mycobacterial DNA is more sensitive and can be used either to verify the presence of organisms seen on special stains or to identify an occult organism. By combining the PCR assays with restriction analyses of the products, it is often possible to speciate the pathogenic organism.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Osaki, M., Adachi, H., Gomyo, Y., Yoshida, H., and Ito, H. (1997) Detection of mycobacterial DNA in formalin-fixed, paraffin-embedded tissue specimens by duplex polymerase chain reaction: application to histopathologic diagnosis. Mod. Pathol. 10, 78ā83.
Totsch, M., Bocker, W., Brommelkamp, E., Fille, M., Kreczy, A., Ofner, D., Schmid, K. W., and Dockhorn-Dworniczak, B. (1996) Diagnostic value of different PCR assays for the detection of mycobacterial DNA in granulomatous lym-phadenopathy. J. Pathol. 178, 221ā226.
Bascunana, C. R. and Belak, K. (1996) Detection and identification of mycobac-teria in formalin-fixed, paraffin-embedded tissues by nested PCR and restriction enzyme analysis. J. Clin. Microbiol. 34, 2351ā2355.
Ferrara, G., Cannone, M., Guadagnino, A., Nappi, O., and Barberis, M. C. (1999) Nested polymerase chain reaction on vaginal smears of tuberculous cervicitis: a case report. Acta Cytol. 43, 308ā312.
Salian, N. V., Rish, J. A., Eisenach, K. D., Cave, M. D., and Bates, J. H. (1998) Polymerase chain reaction to detect Mycobacterium tuberculosis in histologic specimens. Am. J. Respir. Clin. Care Med. 158, 1150ā1155.
Berk, R. H., Yazici, M., Atabey, N., Ozdamar, O. S., Pabuccuoglu, U., and Alici, E. (1996) Detection of Mycobacterium tuberculosis in formaldehyde solution-fixed, paraffin-embedded tissue by polymerase chain reaction in Pottās disease. Spine 21, 1991ā1995.
Marchetti, G., Gori, A., Catozzi, L., Vago, L., Nebuloni, M., Rossi, M. C., Esposti, A. D., Bandera, A., and Franzette, F. (1998) Evaluation of PCR in detection of mycobacterial tuberculosis from formalin-fixed, paraffin-embedded tissues: comparison of four amplification assays. J. Clin. Microbiol. 36, 1512ā1517.
Hardman, W. J., Benian, G. M., Howard, T., McGowan, J. E. Jr., Metchock, B., Murtagh, J. J. (1996) Rapid detection of mycobacteria in inflammatory necrotiz-ing granulomas from formalin-fixed, paraffin-embedded tissue by PCR in clinically high-risk patients with acid-fast stain and culture negative tissue biopsies. Am. J. Clin. Pathol. 106, 184ā389.
Kox, L. F., van Leeuwen, J., Knijper, S., Jansen, H. M., and Kolk, A. H. (1995) PCR assay based on DNA coding for 16S rRNA for detection and identification of mycobacteria in clinical samples. J. Clin. Microbiol. 33, 3225ā3233.
Cook, S. M., Bartos, R. E., Pierson, C. L., and Frank, T. S. (1994) Detection and characterization of atypical mycobacteria by the polymerase chain reaction. Diagn. Mol. Pathol. 3, 53ā58.
Author information
Authors and Affiliations
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
Ā© 2001 Humana Press Inc.
About this protocol
Cite this protocol
Mohl, D., Giordano, T.J. (2001). Detection and Speciation of Mycobacteria in Formalin-Fixed, Paraffin-Embedded Tissue Sections. In: Killeen, A.A. (eds) Molecular Pathology Protocols. Methods in Molecular Medicineā¢, vol 49. Humana Press. https://doi.org/10.1385/1-59259-081-0:379
Download citation
DOI: https://doi.org/10.1385/1-59259-081-0:379
Publisher Name: Humana Press
Print ISBN: 978-0-89603-681-9
Online ISBN: 978-1-59259-081-0
eBook Packages: Springer Protocols