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64 Diagnostic Cytopathology, Vol 8, No I

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Tada M, Omata M, Ohto M. Clinical application of ras gene muta- tion for diagnosis of pancreatic adenocarcinoma. Gastroenterology 199 1; 100:233-8. Shibata D, Almoguera C, Forrester K, Dunitz J, Martin SE. Detec- tion of c-K-ras mutations in fine-needle aspirates from human pan- creatic adenocarcinomas. Cancer Res 1990;50: 1279-83. Shibata D, Nichols, P, Sherrod A, Rabinowits A, Bernstein-Singer L, Hu E. Detection of occult CNS involvement of follicular small cleaved lymphoma by the polymerase chain reaction. Mod Path 1990;3:7 1-5. De Wit D, Steyn L, Shoemaker S , Sogin M. Direct detection of Mycobacterium tuberculosis in clinical specimens by DNA amplifi- cation. J Clin Microbiol 1990;28:243741. Shibata DK, Arnheim N, Martin WJ. Detection of human papil- loma virusin paraffin-embedded tissue using the polymerase chain reaction. J Exp Med 1988;168:255-30. Greer CE, Peterson SL, Kiviat NB, Manos MM. PCR amplification from paraffin-embedded tissues. Am J Clin Pathol 1991;95:124. Battifora H. (letter) Effects of fixatives and fixation times on tissues. Am J Clin Pathol 1991;96:144. McCarthy KP, Sloane JP, Wiedemann LM. Rapid method for dis- tinguishing clonal from polyclonal B cell populations in surgical biopsy specimens. J Clin Pathol 1990;43:429-32. Deane M, Norton JD. Detection of immunoglobulin gene rearrange- ment in B lymphoid malignancies by polymerase chain reaction gene amplification. Br J Haematol 1990; 74:251-6. Bourguin A, Tung R, Galili N, and Sklar J. Rapid, nonradioactive detection of clonal T-cell receptor gene rearrangements in lymphoid neoplasms. Proc Natl Acad Sci USA 1990;87:8536-40. Tyco B., Palmer JD, Link MP, Smith SD, and Sklar J. Polymerase chain reaction amplification of rearranged antigen receptor genes using junction-specific oligonucleotides: possible application for de- tection of minimal residual disease in acute lymphoblastic leukemia. In Cancer cells: molecular diagnostics of human cancer. Cold Spring Harbor, NY: Cold Spring Harbor Laboratory, 1989. Greiner T, Anderson M, Lutz C. Direct sequencing of ALL-specific TCR-gamma genes amplified from archival bone marrow aspirates. Mod Pathol 1991;4:72A. Hanson CA, Holbrook EA, Sheldon S , Schnitzer B, Roth M. Detec- tion of Philadelphia chromosome-positive cells from glass slide smears using the polymerase chain reaction. Am J Pathol 1990; 137:l-6. Lee M, Chang K, Cabanilla F, Freireich El, Trujillo JM, Stass SA. Detection of minimal residual cells carrying the t(14;18) by DNA sequence amplification. Science 1987;237:175-8. Chehab FF, Xiao X, Kan YW, Yen TSB. Detection of cytomegalovirus infection in paraffin-embedded tissue specimens with the polymerase chain reaction. Mod Path 1989;2:75-8. Katzenstein AA. Peiper SC. Detection of Epstein-Barr virus genomes in lymphomatoid granulomatosis: Analysis of 29 cases by the polymerase chain reaction technique. Mod Path 1990;3:43541. Lampertico P. Maker JS, Colombo M, Gerber MA. Detection of hepatitis B virus DNA in formalin-fixed paraffin-embedded liver tissue by the polymerase chain reaction. Am J Pathol. 1990;137: 253-8. Rogers BB, Josephson SL, Mark SK. Detection of herpes simplex virus using the polymerase chain reaction followed by endonuclease cleavage. Am J Pathol 1991;139:1-6. Kwok S, Mack DH, Mullis KB, et al. Identification of human im- muno-deficiency virus sequences by using in vitro enzyme amplifi- cation and oligomer cleavage detection. J Virol 1987;61:1690. Jarrett RF, Clark DA, Josephs SF, Onions DE. Detection of human herpes-virus-6 DNA in peripheral blood and saliva. J Med Virol 1990;32:73-6. Kay R, Komminoth P, Machado M, Wolfe H. Combined polyme- rase chain reaction and in situ hybridisation for the detection of

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single copy genes and viral genomic sequences in intact cells. Mod Pathol 1991;4:124A. Kwok S , Higuchi R. Avoiding false positives with PCR. Nature 1989;339:237-8. Kitchin PA, Szotyori 2, Fromholc C, Almond N. Avoidance of false positives. Nature 1990344:201. Tidy J, Farrell PJ. Retraction: human papillomavirus subtype 16b. Lancet 1989;2:1535. Sarkar G, Sommer S S . Parameters affecting susceptibility of PCR contamination to UV irradiation. Biotechniques 1991; 10591-3. Wright PA, Wynford TD. The polymerase chain reaction: miracle of mirage? A critical review of its uses and limitations in diagnosis and research. J Pathol 1990162:99-117. Erlich HA, Gibbs R, Kazazian HH, eds. Polymerase chain reaction. Cold Spring Harbor, NY: Cold Spring Harbor Laboratory Press, 1989. Innis MA, Gelfand DH, Sninsky JJ, White TJ, eds. PCR Protocols: a guide to methods and applications. San Diego: Academic Press, 1990.

Editorial Comments The polymerase chain reaction and related thermal-cy- cling techniques will have a major impact on the practice of diagnostic pathology in the coming years. Certain cyto- logic procedures, such as the morphologic search for in- fectious agents, might be aided, or replaced by PCR tech- nologies. For example, Wakefield et al. has shown that DNA amplification techniques were superior to silver stains of sputum for the detection of Pneumocystis carinii pneumonia (90% vs. 35%). One can envision a scenario where respiratory cytology specimens are screened with a battery of molecular probes against major pathogens, se- lecting some for further morphological review. Molecular methods can also aid the identification of certain malig- nancies, notably non-Hodgkin’s lymphoma. Molecular di- agnostic procedures, while now labor intensive, will likely by automated in the near future.

As molecular diagnostics are integrated into the diag- nostic laboratory, we must verify that the presence of particular nucleic acids or gene arrangements correlates with clinically significant disease. Thus, as PCR is evalu- ated as an adjunct to cytologic examination the predictive value of a positive result must be considered as a key parameter. It is important for cytopathologists and cyto- technologists to take a leading role in the evaluation of these technologies.

James Linder, M.D.

References 1. Wakefield AE, Guiver L, Miller RF, Hopkin JM. DNA amplifica-

tion on induced sputum samples for diagnosis of Pneumocystis cori- nii pneumonia. Lancet 199 l;337: 1378-9.

2. Schwaiger A, Prior C, Weyrer K, Umlauft F, Gattringer C, Grune- wald K, Totsch M, Fend F. Non-Hodgkin’s lymphoma of the lung diagnosed by gene rearrangement from the bronchoalveolar lavage fluid: A fast and noninvasive method (letter). Blood 1991;77:2538-9.