Analysis of mRNA quality in freshly prepared and archival Papanicolaou samples

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dc.contributor.authorChuaqui, R
dc.contributor.authorCole, K
dc.contributor.authorCuello, M
dc.contributor.authorSilva, M
dc.contributor.authorQuintana, ME
dc.contributor.authorEmmert-Buck, MR
dc.date.accessioned2025-01-21T01:31:53Z
dc.date.available2025-01-21T01:31:53Z
dc.date.issued1999
dc.description.abstractOBJECTIVE: To study the feasibility of utilizing mRNA recovered front cytologic Papanicolaou (Pay) specimens as a resource for gene expression studies of normal and diseased cells.
dc.description.abstractSTUDY DESIGN: To assess the effects of fixation on mRNA recovery and analysis, fresh Pay samples were processed by three separate methods: (1) routine cytologic fixation, (2) 70% ethanol fixation, and (3) air drying without fixation. One-week-old, 1-month-old, 1-year-old and 10-year-old samples were studied to determine the quality of mRNA in archival samples. mRNA quality was analyzed by RT-PCR for the HPRT gene, and by complete transcript amplification. Both heterogeneous (whole slide scrapes) and microdissected cell populations were studied.
dc.description.abstractRESULTS: Reverse transcriptase-polymerase chain reaction (RT-PCR)for the hypoxanthine guanine phosphoribosil transferase gene teas positive in all fresh and archival samples and was not affected by fixative, processing methodology or microdissection. Complete transcript amplification followed by gel electrophoresis showed cDNA smears in all fresh samples with a maximum intensity between 1 and 2 kilobases (kb). Amplification of mRNA was Mot affected by fixation. Smaller cDNA smears were seen in archival specimens with a maximum intensity between 0.5 and 1.5 kb in both one-week-old and one-month-old samples. Smears of approximately 500 base pairs were observed in the 1-year-old and 10-year-old samples. Successful mRNA amplification was possible from microdissected cell populations.
dc.description.abstractCONCLUSION: Messenger RNA recovery and analysis is possible from archival cytologic specimens, suggesting that they call serve as a useful template for RT-PCR analysis of individual genes as well as newly developing high-throughput gene expression methodologies, such as microarrays. Cytologic samples may be particularly useful for study of archival samples as well as diseases from which tissue samples amenable to mRNA-based studies are not available.
dc.fuente.origenWOS
dc.identifier.issn0001-5547
dc.identifier.urihttps://repositorio.uc.cl/handle/11534/97163
dc.identifier.wosidWOS:000082640700015
dc.issue.numero5
dc.language.isoen
dc.pagina.final836
dc.pagina.inicio831
dc.revistaActa cytologica
dc.rightsacceso restringido
dc.subjectPapanicolaou smear
dc.subjectmRNA
dc.subjectgene expression
dc.subjectpolymerase chain reaction
dc.subject.ods03 Good Health and Well-being
dc.subject.odspa03 Salud y bienestar
dc.titleAnalysis of mRNA quality in freshly prepared and archival Papanicolaou samples
dc.typeartículo
dc.volumen43
sipa.indexWOS
sipa.trazabilidadWOS;2025-01-12
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