Isolation of RNA from cell lines and cervical cytology specimens stored in BD SurePath™ preservative fluid and downstream detection of housekeeping gene and HPV E6 expression using real time RT-PCR

Murphy, Patricia; Henderson, Dorian; Adams, Melissa D.; Horlick, Elizabeth A.; Dixon, Eric P.; King, Lorraine; Avissar, Patricia L.; Brown, Charlotte A.; Fischer, Timothy J.; Malinowski, Douglas P.

doi:10.1016/j.jviromet.2008.11.011

Cited by 17 publications

(22 citation statements)

References 29 publications

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“…Noncrosslinking alcoholic reagents yield superior results as RNA fixatives in comparison with aldehydes because they cause little chemical change . RNA quality with both standard and microwave fixation and processing methods has been demonstrated for laboratory‐prepared and commercial alcohol‐based fixatives such as methacarn, ethanol (including a range of concentrations that can vary among different cytologic specimen preparations and within distinct fixative compositions; eg, <24% for SurePath preservative fluid, 60% for Carnoy fixative, 70%, and 95%), FineFix, Carnoy, and PAXgene fixatives, among others . Acceptable preservation of tissue morphology has also been emphasized for some of these fixatives.…”

Section: Rt‐pcr For Fusion Gene Detectionmentioning

confidence: 99%

“…The effects of fixatives on RNA integrity have also been examined in liquid‐based cytology preparations . The ThinPrep technique (Hologic, Inc, Bedford, Mass) applies a methanol‐based fixative called PreservCyt.…”

Section: Rt‐pcr For Fusion Gene Detectionmentioning

confidence: 99%

“…The SurePath method (TriPath Imaging, Burlington, NC) uses an ethanol‐based preservative for the fixation of clinical specimens. Studies examining the effect of SurePath preservation on RNA quality have been conflicting . RNA integrity, evaluated through an analysis of rRNA by Horvath et al, showed severely degraded RNA with poor recovery in a comparison of fixed and unfixed HeLa cells and exfoliated cervical cells.…”

Section: Rt‐pcr For Fusion Gene Detectionmentioning

confidence: 99%

“…Specifically, TRIzol and Qiagen RNeasy protocols do not provide RNA of sufficient quality and quantity from cells stored in SurePath preservative fluid . In contrast, others have demonstrated that RNA isolated from SurePath‐preserved cells is functional in real‐time RT‐PCR assays when an RNA extraction method that incorporates a proteinase K/protease digestion step with temperature modifications is used …”

Section: Rt‐pcr For Fusion Gene Detectionmentioning

confidence: 99%

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Reverse transcription–polymerase chain reaction molecular testing of cytology specimens: Pre‐analytic and analytic factors

Bridge

2016

Cancer Cytopathology

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The introduction of molecular testing into cytopathology laboratory practice has expanded the types of samples considered feasible for identifying genetic alterations that play an essential role in cancer diagnosis and treatment. Reverse transcription–polymerase chain reaction (RT‐PCR), a sensitive and specific technical approach for amplifying a defined segment of RNA after it has been reverse‐transcribed into its DNA complement, is commonly used in clinical practice for the identification of recurrent or tumor‐specific fusion gene events. Real‐time RT‐PCR (quantitative RT‐PCR), a technical variation, also permits the quantitation of products generated during each cycle of the polymerase chain reaction process. This review addresses qualitative and quantitative pre‐analytic and analytic considerations of RT‐PCR as they relate to various cytologic specimens. An understanding of these aspects of genetic testing is central to attaining optimal results in the face of the challenges that cytology specimens may present. Cancer Cytopathol 2017;125:11‐19. © 2016 American Cancer Society.

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Section: Rt‐pcr For Fusion Gene Detectionmentioning

confidence: 99%

Section: Rt‐pcr For Fusion Gene Detectionmentioning

confidence: 99%

Section: Rt‐pcr For Fusion Gene Detectionmentioning

confidence: 99%

Section: Rt‐pcr For Fusion Gene Detectionmentioning

confidence: 99%

See 2 more Smart Citations

Reverse transcription–polymerase chain reaction molecular testing of cytology specimens: Pre‐analytic and analytic factors

Bridge

2016

Cancer Cytopathology

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“…Fixation of cells in SurePath liquid results in degradation of RNA, making it unsuitable for standard real-time PCR amplification (20). Recently, procedures have been proposed that allow real-time PCR amplification and nucleic acid sequence-based amplification (NASBA) using residual material from SurePath preservative fluid (21,22).…”

Section: Nucleic Acid Extraction From Stored Cytology Samplesmentioning

confidence: 99%

Cervical Cytology Biobanking in Europe

Arbyn

Veen²,

Andersson

et al. 2010

Int J Biol Markers

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Int J Biol Markers ( 2010; 3 ): 117-125 25 - 117Original article offers a unique tool for etiological studies, studies on the natural history and molecular pathways of carcinogenesis, evaluation of screening and vaccination effectiveness, and the evaluation of early biomarkers (viral DNA, RNA or proteins, or immunological, genetic or other biological host factors) usable in cancer screening or triage of screen positives. Even when, in the future, detection of nucleic acids of human papillomaviruses would become the standard screening methodology, collection of cellular material will be needed to perform HPV tests. Removal of coverslips from archived conventional Pap smears, extraction of DNA from scraped cells or from microdissected dyskaryotic cells, DNA amplification of human or viral genomic sequences and HPV genotyping have been demonstrated to be feasible in previous studies (2-6). Liquid-based cytology (LBC) is a newer method of preparing cervical cytology samples. Collected cells are transferred in a fluid fixative and evenly dispersed across slides (7). Importantly, the leftover sample contains large amounts of fixed cells with well-preserved morphology, INTRODUCTIONEvery year, tens of millions of cervical cell samples are obtained within the European Union. In countries with well organized cervical cancer screening, the 10-year cumulative attendance rate is well above 90% of all women belonging to the target age range. The average number of smears planned over a lifetime varies among member states: from 7 in Finland and the Netherlands to more than 50 in Germany, Austria and Luxembourg (1). Cervical cell samples are usually stored for 5 to 10 years for medicolegal or quality-control reasons. Certain European laboratories have archived series of Pap smears over several decades. This makes archival cervical samples a valuable resource for research where genetic susceptibility, exposure to external etiological factors as well as outcome (cervical and other malignancies) can be documented through the link with cancer registries.A cervical cytology biobank (CCB) linked to a screening registry, a cancer registry and a HPV vaccination registry

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Biospecimen repositories and cytopathology

Krishnamurthy

2014

Cancer Cytopathology

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Biospecimen repositories are important for the advancement of biomedical research. Literature on the potential for biobanking of fine-needle aspiration, gynecologic, and nongynecologic cytology specimens is very limited. The potential for biobanking of these specimens as valuable additional resources to surgically excised tissues appears to be excellent. The cervicovaginal specimens that can be used for biobanking include Papanicolaou-stained monolayer preparations and residual material from liquidbased cytology preparations. Different types of specimen preparations of fine-needle aspiration and nongynecologic specimens, including Papanicolaou-stained and Diff-Quik-stained smears, cell blocks. and dedicated passes/residual material from fineneedle aspiration stored frozen in a variety of solutions, can be used for biobanking. Because of several gaps in knowledge regarding the standard of operative procedures for the procurement, storage, and quality assessment of cytology specimens, further studies as well as national conferences and workshops are needed not only to create awareness but also to facilitate the use of cytopathology specimens for biobanking. Cancer (Cancer Cytopathol) 2015;123:152-61.

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Isolation of RNA from cell lines and cervical cytology specimens stored in BD SurePath™ preservative fluid and downstream detection of housekeeping gene and HPV E6 expression using real time RT-PCR

Cited by 17 publications

References 29 publications

Reverse transcription–polymerase chain reaction molecular testing of cytology specimens: Pre‐analytic and analytic factors

Reverse transcription–polymerase chain reaction molecular testing of cytology specimens: Pre‐analytic and analytic factors

Cervical Cytology Biobanking in Europe

Biospecimen repositories and cytopathology

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