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Mol Diagn ; 6(2): 125-30, 2001 Jun.
Artículo en Inglés | MEDLINE | ID: mdl-11468697

RESUMEN

BACKGROUND: Molecular diagnosis requires the ability to obtain high-quality nucleic acids that are representative of the disease state. We evaluated the recovery and detection of limiting amounts of viral oncogenic RNA from cells fixed in liquid-based cytology media. METHODS AND RESULTS: Serial dilutions of a human papillomavirus (HPV)-positive cell line fixed in a liquid media was used as a model system. Total nucleic acid (TNA) extraction produced RNA with clearly visible ribosomal bands even after one year of storage. These TNA extracts, treated with DNase-I, were used in an RT-PCR assay for HPV-16 E6-E7 oncogenic transcripts. With chemiluminscent Southern blot detection, samples with one HPV-positive cell in 30,000 were consistently detected. CONCLUSION: PreservCyt-fixed cells can yield RNA suitable for molecular assays even after one year of storage.


Asunto(s)
Química Clínica/métodos , ADN/análisis , ARN Viral/análisis , Proteínas Represoras , Southern Blotting , Electroforesis en Gel de Agar , Femenino , Humanos , Mediciones Luminiscentes , Epidemiología Molecular , Proteínas Oncogénicas Virales/biosíntesis , Papillomaviridae/genética , Proteínas E7 de Papillomavirus , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Sensibilidad y Especificidad , Células Tumorales Cultivadas , Neoplasias del Cuello Uterino/virología
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