<scp>JC</scp> and <scp>BK</scp> virus <scp>DNA</scp> detection in archival slides of urine cytospin from renal transplant patients

Assis, Patricia Gioia de; Carvalho, Carlos Eduardo; Silva, Marcelo Soares; Ribeiro, Bruna dos Santos Paiva; Carvalho, Maria da Glória

doi:10.1111/tid.12901

Cited by 2 publications

(2 citation statements)

References 29 publications

(37 reference statements)

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“…Cytospins are used to concentrate cells from cell suspensions to a designated area where further processing is usually similar to smears. Several types of samples are suitable for cytospin preparations and subsequent molecular analyses 14–16 . Liquid‐based cytology (LBC) samples are immediately fixed cells that are further processed in the laboratory yielding monolayer slide 17 .…”

Section: Introductionmentioning

confidence: 99%

“…Several types of samples are suitable for cytospin preparations and subsequent molecular analyses. 14 , 15 , 16 Liquid‐based cytology (LBC) samples are immediately fixed cells that are further processed in the laboratory yielding monolayer slide. 17 Successful molecular analyses were performed using LBC samples where cells are either scrapped off from slides or used directly as fixed liquid samples.…”

Section: Introductionmentioning

confidence: 99%

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Optimization of pre‐analytical and analytical steps for DNA and RNA analysis of fresh cytology samples

et al. 2022

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Background: Different cytology preparations can be used for molecular diagnostics, however the influence of pre-analytical and analytical steps on the results are not yet well defined. We aimed to determine optimal steps for efficient extraction of DNA and RNA from fresh cells for molecular diagnostics.Methods: MCF7 and FaDu human cell lines, were used as a model to determine fresh cells storage conditions (temperature: 25°C, 4°C, −20°C, −80°C; duration: 0 h, 4 h, 12 h, 24 h, 48 h) and optimal nucleic acids extraction method. Besides, the minimal number of total cells and minimal percentage of mutated cells needed for successful extraction of nucleic acids and subsequent determination of present mutation were evaluated.Results: Extraction of nucleic acids using spin columns yielded the highest quantity and quality of nucleic acids. Isolation of nucleic acids was feasible in all storage conditions, however higher temperature and longer duration of fresh cells storage were associated with lower quality of isolated nucleic acids and similar quantification cycle of housekeeping genes. Successful molecular testing was feasible with least 10 4 cells, while specific mutation was detected in as low as 5% of mutated cells. Conclusions:Our cell line model, mimicking fresh cytology samples, showed that quantity of extracted either DNA or RNA declined with higher temperatures and longer duration of storage but regardless of the storage conditions, we successfully detected both housekeeping genes and mutated gene using qPCR.

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Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Optimization of pre‐analytical and analytical steps for DNA and RNA analysis of fresh cytology samples

et al. 2022

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Sternheimer-Malbin Staining to Detect Decoy Cells in Urine of 213 Kidney Transplant Patients

Yan¹,

Guo²,

Han³

et al. 2020

Transplantation Proceedings

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JC and BK virus DNA detection in archival slides of urine cytospin from renal transplant patients

Abstract: The results highlight the potential use of archival slides of urine cytospin material to differentiate BKV and JCV and demonstrate the importance of improved JCV detection for later kidney transplant recipients.

Cited by 2 publications

References 29 publications

Optimization of pre‐analytical and analytical steps for DNA and RNA analysis of fresh cytology samples

Optimization of pre‐analytical and analytical steps for DNA and RNA analysis of fresh cytology samples

Sternheimer-Malbin Staining to Detect Decoy Cells in Urine of 213 Kidney Transplant Patients

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