ArticleJournal of clinical pathology1994
PCR in situ hybridisation detection of HPV 16 in fixed CaSki and fixed SiHa cell lines.
Article in Journal of clinical pathology, 1994. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 6 papers.
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Who cites it
6 citing papers in PubMed.
- One step DNA amplification of mammalian cells in picoliter microwell arrays.RSC advances · 2019Article
- The potential of RNA as a target for national screening of pre-cancer.Journal of public health in Africa · 2018Article
- In situ analysis of intrahepatic virological events in chronic hepatitis B virus infection.The Journal of clinical investigation · 2016Article
- Article
- Genotypic mapping of HPV and assessment of EBV prevalence in endocervical lesions.Journal of clinical pathology · 1997Review
- Processing of long-stored archival cervical smears for human papillomavirus detection by the polymerase chain reaction.British journal of cancer · 1995Article
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Authors and funding
10 authors.
Funding
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Abstract
aimsTo investigate the feasibility of using fixed cells with the polymerase chain reaction (PCR) in situ hybridisation and to investigate possible reasons for reaction failure.
methodsFixed SiHa and CaSki cells were used in an experimental model of PCR in situ hybridisation for the detection of low and intermediate copy number viral infection in fixed cells.
resultsPCR in situ hybridisation was able to detect one to two copies of human papillomavirus (HPV) 16 in SiHa cells, using small fragment amplicons (120 base pairs), confirming the high detection sensitivity and flexibility of the technique. Problems were encountered with localisation of PCR amplified product in CaSki cells (200-300 copies of HPV 16 per cell) owing to diffusion of product post amplification. Overall, 40% of reactions were successful, which confirms the current unreliability of the technique. Within cell preparations, about 50% of cells contained amplified product.
conclusionPCR in situ hybridisation represents the marriage of two revolutionary molecular pathological techniques. However, it is currently unreliable, with reaction failure common. Standardised, dedicated equipment is urgently required if the technique is to achieve universal acceptance. In the future, the technique may be used to detect chromosomal translocations in human tumours and to study cellular gene expression.
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