[Frontiers in Bioscience 2, c15-29, September 1, 1997]

	[Frontiers in Bioscience 2, c15-29, September 1, 1997 Reprints PubMed CAVEAT LECTOR
	IN SITU PCR. OVERVIEW OF PROCEDURES AND APPLICATIONS Carlos A. Muro-Cacho, M.D., Ph.D. Department of Pathology, H. Lee Moffitt Cancer Center and Research Institute and University of South Florida College of Medicine, Tampa, Florida, USA Received 1/10/97 Accepted 7/15/97 9. REFERENCES 1. Teo IA, Shaunak S. Polymerase chain reaction in situ: an appraisal of an emerging technique. Histochem J 27, 647-659 (1995) 2. Nuovo GJ. PCR in situ Hybridization. Protocols and applications. Ed: Nuovo GJ, Raven Press. New York, 1992 3. Embleton M , Gorochov J, Jones PT, & Winter G. In-cell PCR from mRNA: Amplifying and linking the rearranged immunoglobulin heavy and light chain V-genes within single cells. Nucleic Acids Res 20, 3831-3837 (1992) 4. Patterson B, Till M, Otto P, Goolsby C, Furtado M, McBride L, & Wolinsky S. Detection of HIV-1 DNA and messenger RNA in individual cells by PCR-driven in situ hybridization and flow cytometry. Science 260, 976-979 (1993) 5. Komminoth P, Adams V, Long AA, Roth J, Saremaslani P, Flury R, Schmid M, & Heitz PU. Evaluation of methods for hepatitis C virus (HCV) detection in liver biopsies: comparison of histology, immunohistochemistry, in situ hybridization, reverse transcriptase (RT) PCR, and in situ RT PCR. Path Res Pract 190, 1017-1025 (1994) 6. Chen RH, Fuggle SV. in situ cDNA polymerase chain reaction. A novel technique for detecting mRNA expression. Am J Pathol 143, 1527-1534 (1993) 7. Komminoth P, Long AA. in situ polymerase chain reaction. An overview of methods, applications and limitations of a new molecular technique. Virchows Arch B 64, 67-73 (1993) 8. Long AA, Komminoth P, Lee E, & Wolfe H. Comparison of indirect and direct in situ polymerase chain reaction in cell preparations and tissue sections. Detection of viral DNA, gene rearrangements and chromosomal translocations. Histochemistry 99, 151-162 (1993) 9. Nuovo G. Analysis of nonspecific DNA synthesis during in situ PCR and solution-phase PCR. PCR Methods Appl 4, 89-96 (1994) 10. Pestaner C, Bibbo M, Bobroski L, Seshamma T, & Bagasra O. Potential of the in situ Polymerase Chain Reaction in diagnostic cytology. Acta Cytologica 38, 676-680, (1994) 11. Tsongalis GJ, McPhail AH, Lodge RRD, Chapman JF, & Silverman LM. Localized in situ amplification (LISA): a novel approach to in situ PCR. Clin Chem 40, 381-384 (1994) 12. Yap EP, McGee JO. Slide PCR: DNA amplification from cell samples on microscopic glass slides. Nucleic Acids Res 19, 4294-4298 (1991) 13. Komminoth P, Long AA, Ray R, & Wolfe HJ. in situ polymerase chain reaction detection of viral DNA, single copy genes and gene rearrangements in cell suspensions and cytospins. Diagn Mol Pathol 1, 85-97 (1992) 14. Haase AT, Retzel EF, & Staskus KA. Amplification and detection of lentiviral DNA inside cells. Proc Natl Acad Sci USA 87, 4971-4975 (1990) 15. Bagasra O, Sheshamma T, & Pomeranz RJ. Polymerase chain reaction in situ: Intracellular amplification and detection of HIV-1 proviral DNA and other specific genes. J Immunol Methods 158, 131-145 (1993) 16. Chiu KP, Cohen S, Morris D., & Jordan G. Intracellular amplification of proviral DNA in tissue sections using the polymerase chain reaction. J Histochem Biochem 40, 333-341 (1992) 17. Greer CE. PCR amplification from paraffin-embedded tissues: recommendations on fixatives for long-term storage and prospective studies. PCR Methods Appl 95, 117-124 (1991) 18. Greer CE, Petterson SL, Kiviat NB, & Manos MM. PCR amplification from paraffin-embedded tissues: effects of fixative and fixative times. Am J Clin Pathol 32, 85-112 (1991) 19. O'Leary JJ, Browne G, Landers RJ, Crowley M, Healy IB, Street JT, Pollock AM, Murphy J, Johnson MI, & Lewis FA. The importance of fixation procedures on DNA template and its suitability for solution-phase polymerase chain reaction and PCR in situ hybridization. Histochem J 26, 337-346 (1994) 20. Ben-Ezara J, Johnson DA, Rossi J, Cook N, & Wu A. Effect of fixation on the amplification of nucleic acids from paraffin-embedded material by the polymerase chain reaction. J Histochem Cytochem 39, 351-354 (1991) 21. Embretson J, Zupancic M, Beneke J, Till M, Wolinsky S, Ribas J, Burke A, & Haase A. Analysis of human immunodeficiency virus-infected tissues by amplification and in situ hybridization reveals latent and permissive infections at single cell resolution. Proc Natl Acad Sci USA 90, 357-361 (1993) 22. Griffin HG, Griffin AM. PCR Technology Current Innovations. Eds. Griffin HG, and Griffin AM CRC Press INC. 1994 23. White BA. PCR Protocols, Current Methods and Applications. Ed. White BA. Humana Press Inc, 1993 24. Wu DY, Ugozzoli L, Pal BK, Qian J, & Wallace B. The effect of temperature and oligonucleotide primer length on the specificity and efficiency of amplification by the polymerase chain reaction. DNA and Cell Biology 10, 233-238 (1991) 25. Teo IA, Shaunak S. PCR in situ: aspects which reduce amplification and generate false-positive results. Histochem J 27, 660-669 (1995) 26. Rychlik W, Rhoads RE. A computer program for choosing optimal oligonucleotides for filter hybridization, sequencing and for in vitro amplification of DNA. Nucleic Acids Res18, 6409-6412 (1990) 27. Wetmur JG, Davidson N. Kinetics of renaturation of DNA. J Mol Biol 31, 349-370 (1986) 28. Landegren U. Molecular mechanisms of nucleic acid sequence amplification. Trends Genet 9, 199-204 (1993) 29. Ponce MR, Micol JL. PCR amplification of long DNA fragments. Nucleic Acids Res 20, 623-625 (1995) 30. Saiki RK, Gelfand DH, Stoffel S, Scharf SJ, Higuchi R, Horn GT, Mullis KB, & Erlich HA. Primer directed enzymatic amplification of DNA with a thermostable DNA polymerase. Science, 239, 487-491 (1988) 31. Long AA, Komminoth P. in situ PCR. An overview. In PRINS and in situ PCR Protocols. Methods in Molecular Biology Vol. 71. Ed. Gosden JR, Humana Press, Totowa, New Jersey, USA, 1997, pgs.141-161 32. Martinez A, Cuttitta F. Bringing together morphologists and molecular biologists: in situ PCR and RT-PCR. The NIH Catalyst 3, 12-22 (1995) 33. Nuovo G, Becker J, Margiotta M, & McConnell P. Histological distribution of polymerase chain reaction-amplified human papillomavirus 6 and 11 DNA in penile lesions. Am J Surg Pathol 16, 269-275 (1992) 34. Bernard C, Mougin C, Bettinger D, Didier JM, & Lab M. Detection of human papilloma virus by in situ polymerase chain reaction in paraffin-embedded cervical biopsies. Mol Cell Probes 8, 337-343 (1994) 35. Sallstrom JF, Zehbe I, Alemi M, & Wilander E. Pitfalls of in situ polymerase chain reaction (PCR) using direct incorporation of labelled nucleotides. Anticancer Res 13, 1153-1157 (1993) 36. Komminoth P, Long AA. Questioning in-situ PCR. Detection of epidermal growth factor receptor mRNA in tissue sections from biopsy specimens using in-situ polymerase chain reaction (letter). Am J Pathol 145, 742-748 (1994) 37. Hsu SM, Raine L, & Fanger HA. Use of Avidin-Biotin-Peroxidase Complex (ABC) in immunoperoxidase techniques: a comparison of ABC and unlabeled antibody (PAP) procedure. J Histochem Cytochem 29, 577-581 (1981) 38. Sallstrom JF, Alemi M, Spets H, & Zehbe I. Nonspecific amplification in in-situ PCR by direct incorporation of reporter molecules. Cell Vision 1, 243-251 (1994) 39. Zehbe I, Sallstrom JF, Hacker GW, Hauser-kronberger C, Rylander E, & Wilander E. Indirect and direct in situ PCR for the detection of human papillomavirus. An evaluation of two methods and a double staining technique. Cell Vision 1, 163-167 (1994) 40. Staecker H, Cammer H, Rubinstein R, & VandeWater T. A procedure for RT-PCR amplification of mRNAs on histological specimens. Biotechniques 6, 76-80 (1994) 41. Patel V, Shum-Siu A, Heniford B, Wieman T, & Hendler F. Detection of epidermal growth factor receptor mRNA in tissue sections from biopsy specimens using in situ polymerase chain reaction. Am J Pathol 144, 7-14 (1994) 42. Martinez A, Cuttitta F. Non-radioactive Localization of Nucleic Acids by Direct in situ PCR and in situ RT-PCR in Paraffin-embedded Sections. J Histochem Cytochem 43, 739-747 (1995) 43. Crisan D, Mattson JC. Retrospective DNA analysis using fixed tissue specimens. DNA Cell Biol 2, 455-464 (1993) 44. Grafstrom RC, Fornace AJR, & Harris CC. Repair of DNA damage caused by formaldehyde in human cells. Cancer Res 44, 4323-432 (1984) 45. Sallstrom J, Zehbe I, Alemi M, & Wilander E. Pitfalls of in situ polymerase chain reaciton (PCR) using direct incorporation of labelled nucleotides. Anticancer Res 13, 1153-1158 (1993) 46. Kellog DE, Rybalkin I, Chen S, Mukhamedong N, Vlasik T, Siebert PD, & Chenchik A. Taq Start antibody: "hot start" PCR facilitated by a neutralizing monoclonal antibody directed against Taq DNA Polymerase. BioTechniques 6, 1134-1137 (1994) 47. Nuovo G, Gallery F, Hom R, MacConnell P, & Bloch W. Importance of different variables for enhancing in situ detection of PCR-amplified DNA. PCR Methods Appl 2, 305-312 (1995) 48. Zaki SR, Heneine W, Coffield LM, Greer PW, Sinha SD, & Folks TM. In-situ polymerase chain reaction: applications and current limitations. AIDS 8, 1186-1188 (1994) 49. Bagasra O, Hauptman SP, Lischner HW, Sachs M, & Pomerantz RJ. Detection of human immunodeficiency virus type 1 provirus in mononuclear cells by in situ polymerase chain reaction. N Engl J Med 326, 1385-1391 (1992) 50. Uhl GR. A review of in situ Hybridization technique: relevance to combined immunocytochemical studies. In: Immunohistochemistry II. Ed. A. C. Cuello, John Wiley and Sons, 1993 51. Zehbe I, Hacker G, Rylander E, Sallstrom J, & Wilander E. Detection of single HPV copies in SiHa cells by in situ polymerase chain reaction (in situ PCR) combined with immunoperoxidase and immunogold-silver staining (IGSS) techniques. Anticancer Res 12, 2165-2168 (1992) 52. Chiu KP, Cohen SH, Morris DW, & Jordan GW. Intracellular amplification of proviral DNA in tissue sections using the polymerase chain reaction. J Histochem Cytochem 240, 333-341 (1992) 53. Patterson B, Till M, Otto P, Goolsby C, Furtado M, MacBride L, & Wolinsky S. Detection of HIV-1 DNA and messenger RNA in individual cells by PCR-driven in situ hybridization and flow cytometry. Science 260, 976-979 (1993) 54. Heniford BW. Variation in cellular EGF receptor mRNA expression demonstrated by in situ reverse transcriptase polymerase chain reaction. Nucleic Acids Res 21, 3159-3166 (1993) 55. Spann W, Pachmann K, Zabnieska H, Pielmeir A, & Emmerich. in situ amplification of single copy gene segments in individual cells by the polymerase chain reaction. Infection 19, 242-244 (1991) 56. Staskus K, Couch L, Bitterman P, Retzel E, Zupancic M, List J, & Haase A. in situ amplification of visna virus DNA in tissue sections reveals a reservoir of latently infected cells. Microbiol Pathog 11, 67-76 (1991) 57. Ueki K, Richardson EP, Henson JW, & Louis DN. in situ polymerase chain reaction demonstration of JC Virus in progressive multifocal leukoencephalopathy, including an index case. Annals Neurol 36, 670-673 (1994) 58. Mehta A. Detection of herpes simplex virus sequences in the trigeminal ganglia of latently infected mice by an in situ DNA PCR method. Cell Vision 1, 110-115 (1994) 59. Martinez A, Miller MJ, & Cuttita F. Detection of insulin growth factor-1 (IGF-1) by in situ RT-amplification. Proceedings of the Third Analytical Morphology Meeting Ed. Gu J. Atlantic City, NJ, USA, 1995 60. Koch J, Hindkjaer J, Mogensen J, Kolvraa S, & Bolund L. An improved method for chromosome-specific labeling of alpha satellite DNA in situ by using denatured double-stranded DNA probes as primers in a primed in situ labeling (PRINS) procedure. Genet Anal Tech Appl 8, 171-178 (1991) 61. Gosden J, Hanratty D, Startling J, Fantes J, Mitchell A, & Porteous D. Oligonucleotide primed in situ DNA synthesis (PRINS): a method for chromosome mapping, banding, and investigation of sequence organization. Cytogenet Cell Genet 57, 100-104 (1991) 62. Gosden J, Hanratty D. PCR in situ: A rapid alternative to in situ hybridization for mapping short, low copy number sequences without isotopes. BioTechniques 15, 78-80 (1993) 63. Gingeras TR, Richman DD, Kwoh DY, & Guatelli JC. Use of self-sustained sequence replication amplification reaction to analyze and detect mutations in zidovine-resistant human immunodeficiency virus. J Infect Dis 164, 1066-1074 (1991) 64. Guatelli JC, Whitfield KM, Kwoh DY, Baringer KJ, Richman DD, & Gingeras TR. Isothermal in vitro amplification of nucleic acids by a multienzyme reaction modeled after retroviral replication. Proc Natl Acad Sci USA 7, 1874-1878 (1994)

[Frontiers in Bioscience 2, c15-29, September 1, 1997
Reprints
PubMed
CAVEAT LECTOR

IN SITU PCR. OVERVIEW OF PROCEDURES AND APPLICATIONS

Carlos A. Muro-Cacho, M.D., Ph.D.

Department of Pathology, H. Lee Moffitt Cancer Center and Research Institute and University of South Florida College of Medicine, Tampa, Florida, USA

Received 1/10/97 Accepted 7/15/97

9. REFERENCES

1. Teo IA, Shaunak S. Polymerase chain reaction in situ: an appraisal of an emerging technique. Histochem J 27, 647-659 (1995)

2. Nuovo GJ. PCR in situ Hybridization. Protocols and applications. Ed: Nuovo GJ, Raven Press. New York, 1992

3. Embleton M , Gorochov J, Jones PT, & Winter G. In-cell PCR from mRNA: Amplifying and linking the rearranged immunoglobulin heavy and light chain V-genes within single cells. Nucleic Acids Res 20, 3831-3837 (1992)

4. Patterson B, Till M, Otto P, Goolsby C, Furtado M, McBride L, & Wolinsky S. Detection of HIV-1 DNA and messenger RNA in individual cells by PCR-driven in situ hybridization and flow cytometry. Science 260, 976-979 (1993)

5. Komminoth P, Adams V, Long AA, Roth J, Saremaslani P, Flury R, Schmid M, & Heitz PU. Evaluation of methods for hepatitis C virus (HCV) detection in liver biopsies: comparison of histology, immunohistochemistry, in situ hybridization, reverse transcriptase (RT) PCR, and in situ RT PCR. Path Res Pract 190, 1017-1025 (1994)

6. Chen RH, Fuggle SV. in situ cDNA polymerase chain reaction. A novel technique for detecting mRNA expression. Am J Pathol 143, 1527-1534 (1993)

7. Komminoth P, Long AA. in situ polymerase chain reaction. An overview of methods, applications and limitations of a new molecular technique. Virchows Arch B 64, 67-73 (1993)

8. Long AA, Komminoth P, Lee E, & Wolfe H. Comparison of indirect and direct in situ polymerase chain reaction in cell preparations and tissue sections. Detection of viral DNA, gene rearrangements and chromosomal translocations. Histochemistry 99, 151-162 (1993)

9. Nuovo G. Analysis of nonspecific DNA synthesis during in situ PCR and solution-phase PCR. PCR Methods Appl 4, 89-96 (1994)

10. Pestaner C, Bibbo M, Bobroski L, Seshamma T, & Bagasra O. Potential of the in situ Polymerase Chain Reaction in diagnostic cytology. Acta Cytologica 38, 676-680, (1994)

11. Tsongalis GJ, McPhail AH, Lodge RRD, Chapman JF, & Silverman LM. Localized in situ amplification (LISA): a novel approach to in situ PCR. Clin Chem 40, 381-384 (1994)

12. Yap EP, McGee JO. Slide PCR: DNA amplification from cell samples on microscopic glass slides. Nucleic Acids Res 19, 4294-4298 (1991)

13. Komminoth P, Long AA, Ray R, & Wolfe HJ. in situ polymerase chain reaction detection of viral DNA, single copy genes and gene rearrangements in cell suspensions and cytospins. Diagn Mol Pathol 1, 85-97 (1992)

14. Haase AT, Retzel EF, & Staskus KA. Amplification and detection of lentiviral DNA inside cells. Proc Natl Acad Sci USA 87, 4971-4975 (1990)

15. Bagasra O, Sheshamma T, & Pomeranz RJ. Polymerase chain reaction in situ: Intracellular amplification and detection of HIV-1 proviral DNA and other specific genes. J Immunol Methods 158, 131-145 (1993)

16. Chiu KP, Cohen S, Morris D., & Jordan G. Intracellular amplification of proviral DNA in tissue sections using the polymerase chain reaction. J Histochem Biochem 40, 333-341 (1992)

17. Greer CE. PCR amplification from paraffin-embedded tissues: recommendations on fixatives for long-term storage and prospective studies. PCR Methods Appl 95, 117-124 (1991)

18. Greer CE, Petterson SL, Kiviat NB, & Manos MM. PCR amplification from paraffin-embedded tissues: effects of fixative and fixative times. Am J Clin Pathol 32, 85-112 (1991)

19. O'Leary JJ, Browne G, Landers RJ, Crowley M, Healy IB, Street JT, Pollock AM, Murphy J, Johnson MI, & Lewis FA. The importance of fixation procedures on DNA template and its suitability for solution-phase polymerase chain reaction and PCR in situ hybridization. Histochem J 26, 337-346 (1994)

20. Ben-Ezara J, Johnson DA, Rossi J, Cook N, & Wu A. Effect of fixation on the amplification of nucleic acids from paraffin-embedded material by the polymerase chain reaction. J Histochem Cytochem 39, 351-354 (1991)

21. Embretson J, Zupancic M, Beneke J, Till M, Wolinsky S, Ribas J, Burke A, & Haase A. Analysis of human immunodeficiency virus-infected tissues by amplification and in situ hybridization reveals latent and permissive infections at single cell resolution. Proc Natl Acad Sci USA 90, 357-361 (1993)

22. Griffin HG, Griffin AM. PCR Technology Current Innovations. Eds. Griffin HG, and Griffin AM CRC Press INC. 1994

23. White BA. PCR Protocols, Current Methods and Applications. Ed. White BA. Humana Press Inc, 1993

24. Wu DY, Ugozzoli L, Pal BK, Qian J, & Wallace B. The effect of temperature and oligonucleotide primer length on the specificity and efficiency of amplification by the polymerase chain reaction. DNA and Cell Biology 10, 233-238 (1991)

25. Teo IA, Shaunak S. PCR in situ: aspects which reduce amplification and generate false-positive results. Histochem J 27, 660-669 (1995)

26. Rychlik W, Rhoads RE. A computer program for choosing optimal oligonucleotides for filter hybridization, sequencing and for in vitro amplification of DNA. Nucleic Acids Res18, 6409-6412 (1990)

27. Wetmur JG, Davidson N. Kinetics of renaturation of DNA. J Mol Biol 31, 349-370 (1986)

28. Landegren U. Molecular mechanisms of nucleic acid sequence amplification. Trends Genet 9, 199-204 (1993)

29. Ponce MR, Micol JL. PCR amplification of long DNA fragments. Nucleic Acids Res 20, 623-625 (1995)

30. Saiki RK, Gelfand DH, Stoffel S, Scharf SJ, Higuchi R, Horn GT, Mullis KB, & Erlich HA. Primer directed enzymatic amplification of DNA with a thermostable DNA polymerase. Science, 239, 487-491 (1988)

31. Long AA, Komminoth P. in situ PCR. An overview. In PRINS and in situ PCR Protocols. Methods in Molecular Biology Vol. 71. Ed. Gosden JR, Humana Press, Totowa, New Jersey, USA, 1997, pgs.141-161

32. Martinez A, Cuttitta F. Bringing together morphologists and molecular biologists: in situ PCR and RT-PCR. The NIH Catalyst 3, 12-22 (1995)

33. Nuovo G, Becker J, Margiotta M, & McConnell P. Histological distribution of polymerase chain reaction-amplified human papillomavirus 6 and 11 DNA in penile lesions. Am J Surg Pathol 16, 269-275 (1992)

34. Bernard C, Mougin C, Bettinger D, Didier JM, & Lab M. Detection of human papilloma virus by in situ polymerase chain reaction in paraffin-embedded cervical biopsies. Mol Cell Probes 8, 337-343 (1994)

35. Sallstrom JF, Zehbe I, Alemi M, & Wilander E. Pitfalls of in situ polymerase chain reaction (PCR) using direct incorporation of labelled nucleotides. Anticancer Res 13, 1153-1157 (1993)

36. Komminoth P, Long AA. Questioning in-situ PCR. Detection of epidermal growth factor receptor mRNA in tissue sections from biopsy specimens using in-situ polymerase chain reaction (letter). Am J Pathol 145, 742-748 (1994)

37. Hsu SM, Raine L, & Fanger HA. Use of Avidin-Biotin-Peroxidase Complex (ABC) in immunoperoxidase techniques: a comparison of ABC and unlabeled antibody (PAP) procedure. J Histochem Cytochem 29, 577-581 (1981)

38. Sallstrom JF, Alemi M, Spets H, & Zehbe I. Nonspecific amplification in in-situ PCR by direct incorporation of reporter molecules. Cell Vision 1, 243-251 (1994)

39. Zehbe I, Sallstrom JF, Hacker GW, Hauser-kronberger C, Rylander E, & Wilander E. Indirect and direct in situ PCR for the detection of human papillomavirus. An evaluation of two methods and a double staining technique. Cell Vision 1, 163-167 (1994)

40. Staecker H, Cammer H, Rubinstein R, & VandeWater T. A procedure for RT-PCR amplification of mRNAs on histological specimens. Biotechniques 6, 76-80 (1994)

41. Patel V, Shum-Siu A, Heniford B, Wieman T, & Hendler F. Detection of epidermal growth factor receptor mRNA in tissue sections from biopsy specimens using in situ polymerase chain reaction. Am J Pathol 144, 7-14 (1994)

42. Martinez A, Cuttitta F. Non-radioactive Localization of Nucleic Acids by Direct in situ PCR and in situ RT-PCR in Paraffin-embedded Sections. J Histochem Cytochem 43, 739-747 (1995)

43. Crisan D, Mattson JC. Retrospective DNA analysis using fixed tissue specimens. DNA Cell Biol 2, 455-464 (1993)

44. Grafstrom RC, Fornace AJR, & Harris CC. Repair of DNA damage caused by formaldehyde in human cells. Cancer Res 44, 4323-432 (1984)

45. Sallstrom J, Zehbe I, Alemi M, & Wilander E. Pitfalls of in situ polymerase chain reaciton (PCR) using direct incorporation of labelled nucleotides. Anticancer Res 13, 1153-1158 (1993)

46. Kellog DE, Rybalkin I, Chen S, Mukhamedong N, Vlasik T, Siebert PD, & Chenchik A. Taq Start antibody: "hot start" PCR facilitated by a neutralizing monoclonal antibody directed against Taq DNA Polymerase. BioTechniques 6, 1134-1137 (1994)

47. Nuovo G, Gallery F, Hom R, MacConnell P, & Bloch W. Importance of different variables for enhancing in situ detection of PCR-amplified DNA. PCR Methods Appl 2, 305-312 (1995)

48. Zaki SR, Heneine W, Coffield LM, Greer PW, Sinha SD, & Folks TM. In-situ polymerase chain reaction: applications and current limitations. AIDS 8, 1186-1188 (1994)

49. Bagasra O, Hauptman SP, Lischner HW, Sachs M, & Pomerantz RJ. Detection of human immunodeficiency virus type 1 provirus in mononuclear cells by in situ polymerase chain reaction. N Engl J Med 326, 1385-1391 (1992)

50. Uhl GR. A review of in situ Hybridization technique: relevance to combined immunocytochemical studies. In: Immunohistochemistry II. Ed. A. C. Cuello, John Wiley and Sons, 1993

51. Zehbe I, Hacker G, Rylander E, Sallstrom J, & Wilander E. Detection of single HPV copies in SiHa cells by in situ polymerase chain reaction (in situ PCR) combined with immunoperoxidase and immunogold-silver staining (IGSS) techniques. Anticancer Res 12, 2165-2168 (1992)

52. Chiu KP, Cohen SH, Morris DW, & Jordan GW. Intracellular amplification of proviral DNA in tissue sections using the polymerase chain reaction. J Histochem Cytochem 240, 333-341 (1992)

53. Patterson B, Till M, Otto P, Goolsby C, Furtado M, MacBride L, & Wolinsky S. Detection of HIV-1 DNA and messenger RNA in individual cells by PCR-driven in situ hybridization and flow cytometry. Science 260, 976-979 (1993)

54. Heniford BW. Variation in cellular EGF receptor mRNA expression demonstrated by in situ reverse transcriptase polymerase chain reaction. Nucleic Acids Res 21, 3159-3166 (1993)

55. Spann W, Pachmann K, Zabnieska H, Pielmeir A, & Emmerich. in situ amplification of single copy gene segments in individual cells by the polymerase chain reaction. Infection 19, 242-244 (1991)

56. Staskus K, Couch L, Bitterman P, Retzel E, Zupancic M, List J, & Haase A. in situ amplification of visna virus DNA in tissue sections reveals a reservoir of latently infected cells. Microbiol Pathog 11, 67-76 (1991)

57. Ueki K, Richardson EP, Henson JW, & Louis DN. in situ polymerase chain reaction demonstration of JC Virus in progressive multifocal leukoencephalopathy, including an index case. Annals Neurol 36, 670-673 (1994)

58. Mehta A. Detection of herpes simplex virus sequences in the trigeminal ganglia of latently infected mice by an in situ DNA PCR method. Cell Vision 1, 110-115 (1994)

59. Martinez A, Miller MJ, & Cuttita F. Detection of insulin growth factor-1 (IGF-1) by in situ RT-amplification. Proceedings of the Third Analytical Morphology Meeting Ed. Gu J. Atlantic City, NJ, USA, 1995

60. Koch J, Hindkjaer J, Mogensen J, Kolvraa S, & Bolund L. An improved method for chromosome-specific labeling of alpha satellite DNA in situ by using denatured double-stranded DNA probes as primers in a primed in situ labeling (PRINS) procedure. Genet Anal Tech Appl 8, 171-178 (1991)

61. Gosden J, Hanratty D, Startling J, Fantes J, Mitchell A, & Porteous D. Oligonucleotide primed in situ DNA synthesis (PRINS): a method for chromosome mapping, banding, and investigation of sequence organization. Cytogenet Cell Genet 57, 100-104 (1991)

62. Gosden J, Hanratty D. PCR in situ: A rapid alternative to in situ hybridization for mapping short, low copy number sequences without isotopes. BioTechniques 15, 78-80 (1993)

63. Gingeras TR, Richman DD, Kwoh DY, & Guatelli JC. Use of self-sustained sequence replication amplification reaction to analyze and detect mutations in zidovine-resistant human immunodeficiency virus. J Infect Dis 164, 1066-1074 (1991)

64. Guatelli JC, Whitfield KM, Kwoh DY, Baringer KJ, Richman DD, & Gingeras TR. Isothermal in vitro amplification of nucleic acids by a multienzyme reaction modeled after retroviral replication. Proc Natl Acad Sci USA 7, 1874-1878 (1994)