ArticleHistochemistry1993
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.
Article in Histochemistry, 1993. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 26 papers.
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Who cites it
26 citing papers in PubMed, 151 citations in OpenAlex.
- Results of MDR-1 vector modification trial indicate that granulocyte/macrophage colony-forming unit cells do not contribute to posttransplant hematopoietic recovery following intensive systemic therapy.Proceedings of the National Academy of Sciences of the United States of America · 1996Trial
- Nanobarcoding: detecting nanoparticles in biological samples using in situ polymerase chain reaction.International journal of nanomedicine · 2012Article
- Streptococcus pneumoniae as a frequent cause of severe community-acquired pneumonia among children in Beijing.European journal of clinical microbiology & infectious diseases : official publication of the European Society of Clinical Microbiology · 2009Article
- Improvement of in situ PCR by optimization of PCR cycle number and proteinase k concentration: localization of x chromosome-linked phosphoglycerate kinase-1 gene in mouse reproductive organs.Acta histochemica et cytochemica · 2009Article
- Topographic and quantitative display of integrated human immunodeficiency virus-1 provirus DNA in human lymph nodes by real-time polymerase chain reaction.The Journal of molecular diagnostics : JMD · 2005Article
- Strategies for signal amplification in nucleic acid detection.Molecular biotechnology · 2001Review
- Quantitative analysis of latent human cytomegalovirus.Journal of virology · 1999Article
- CARD In Situ Hybridization: Sights and Signals.Endocrine pathology · 1999Article
- In cell amplification.Molecular pathology : MP · 1998Review
- Development of a direct in situ PCR method for detection of specific bacteria in natural environments.Applied and environmental microbiology · 1998Article
- Detection of HTLV-1 by polymerase chain reaction in situ hybridization in adult T-cell leukemia/lymphoma.The American journal of pathology · 1998Article
- Nonradioactive In Situ Hybridization: Recent Techniques and Applications.Endocrine pathology · 1998Article
- In situ polymerase chain reaction-based localization studies support role of human herpesvirus-8 as the cause of two AIDS-related neoplasms: Kaposi's sarcoma and body cavity lymphoma.The Journal of clinical investigation · 1997Article
- Sensitive in situ hybridization with catalyzed reporter deposition, streptavidin-Nanogold, and silver acetate autometallography: detection of single-copy human papillomavirus.The American journal of pathology · 1997Article
- In situ hybridization: methods and applications.Journal of clinical laboratory analysis · 1997Review
- Direct in situ nucleic acid amplification: control of artefact and use of labelled primers.Clinical molecular pathology · 1996Article
- Rapid isolation of recombinant lambda phage DNA for use in fluorescence in situ hybridization.Chromosome research : an international journal on the molecular, supramolecular and evolutionary aspects of chromosome biology · 1996Article
- Fluorescence in situ hybridization with cosmid clones for the detection of human cytomegalovirus DNA in peripheral blood leukocytes.Histochemistry and cell biology · 1996Article
- Applications of immunogold and lectin-gold labeling in tumor research and diagnosis.Histochemistry and cell biology · 1996Review
- PCR-in situ hybridization detection of human T-cell lymphotropic virus type 1 (HTLV-1) tax proviral DNA in peripheral blood lymphocytes of patients with HTLV-1-associated neurologic disease.Journal of virology · 1996Article
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Authors and funding
4 authors at 2 institutions in 2 countries.
Funding
No grant is acknowledged in the PubMed record.
Abstract
Different approaches to the in-situ polymerase chain reaction (in-situ PCR) were compared in the detection and in-situ localization of chromosomal translocations (t14; 18) immunoglobulin gene rearrangements and viral DNA (cytomegalovirus, hepatitis B-virus) in cell suspensions, cytospins and tissue sections. Single and multiple primer pairs were compared in the amplification step of indirect in-situ PCR and long genomic probes or internal oligonucleotide probes in the subsequent in-situ hybridization (ISH). For direct in-situ PCR, in which amplification products were directly labeled with digoxigenin-11-dUTP during PCR and detected immunohistochemically, only single primer pairs were used for amplification. In-situ PCR yielded best results in the cell suspensions and worked less efficiently in cytospins or tissue sections. Quantification of the results obtained in artificial cell mixtures yielded only an approximate correlation between the number of expected and observed positive cells. The specificity of the results was greater with indirect in-situ PCR than direct in-situ PCR, where false positive results were frequent. Successful indirect in-situ PCR in tissue sections required the use of multiple primer pairs for amplification and genomic probes for detection by ISH. False positive results in direct in-situ PCR were caused by primer-independent, but DNA polymerase- and cycling-dependent incorporation of digoxigenin-labeled nucleotides into cellular DNA, possibly related to DNA repair and/or internal priming. Non-specific results were most marked in tissue sections and were much less frequent in cell suspensions. In-situ PCR includes a number of different techniques, which are not equally applicable to different starting materials. Accurate interpretation of the results requires vigorous controls.
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