Detection of VAR2CSA‐captured colorectal cancer cells from blood samples by real‐time reverse transcription PCR

Research output: Contribution to journal › Journal article › Research › peer-review

Standard

Detection of VAR2CSA‐captured colorectal cancer cells from blood samples by real‐time reverse transcription PCR. / Bang‐christensen, Sara R.; Katerov, Viatcheslav; Jørgensen, Amalie M.; Gustavsson, Tobias; Choudhary, Swati; Theander, Thor G.; Salanti, Ali; Allawi, Hatim T.; Agerbæk, Mette.

In: Cancers, Vol. 13, No. 23, 5881, 2021.

Research output: Contribution to journal › Journal article › Research › peer-review

Harvard

Bang‐christensen, SR, Katerov, V, Jørgensen, AM, Gustavsson, T, Choudhary, S, Theander, TG, Salanti, A, Allawi, HT & Agerbæk, M 2021, 'Detection of VAR2CSA‐captured colorectal cancer cells from blood samples by real‐time reverse transcription PCR', Cancers, vol. 13, no. 23, 5881. https://doi.org/10.3390/cancers13235881

APA

Bang‐christensen, S. R., Katerov, V., Jørgensen, A. M., Gustavsson, T., Choudhary, S., Theander, T. G., Salanti, A., Allawi, H. T., & Agerbæk, M. (2021). Detection of VAR2CSA‐captured colorectal cancer cells from blood samples by real‐time reverse transcription PCR. Cancers, 13(23), [5881]. https://doi.org/10.3390/cancers13235881

Vancouver

Bang‐christensen SR, Katerov V, Jørgensen AM, Gustavsson T, Choudhary S, Theander TG et al. Detection of VAR2CSA‐captured colorectal cancer cells from blood samples by real‐time reverse transcription PCR. Cancers. 2021;13(23). 5881. https://doi.org/10.3390/cancers13235881

Author

Bang‐christensen, Sara R. ; Katerov, Viatcheslav ; Jørgensen, Amalie M. ; Gustavsson, Tobias ; Choudhary, Swati ; Theander, Thor G. ; Salanti, Ali ; Allawi, Hatim T. ; Agerbæk, Mette. / Detection of VAR2CSA‐captured colorectal cancer cells from blood samples by real‐time reverse transcription PCR. In: Cancers. 2021 ; Vol. 13, No. 23.

Bibtex

@article{9609b2cff1f8414fa579835121603d75,
title = "Detection of VAR2CSA‐captured colorectal cancer cells from blood samples by real‐time reverse transcription PCR",
abstract = "Analysis of circulating tumor cells (CTCs) from blood samples provides a non‐invasive approach for early cancer detection. However, the rarity of CTCs makes it challenging to establish assays with the required sensitivity and specificity. We combine a highly sensitive CTC capture assay exploiting the cancer cell binding recombinant malaria VAR2CSA protein (rVAR2) with the detection of colon‐related mRNA transcripts (USH1C and CKMT1A). Cancer cell transcripts are detected by RT‐qPCR using proprietary Target Enrichment Long‐probe Quantitative Amplified Signal (TELQAS) technology. We validate each step of the workflow using colorectal cancer (CRC) cell lines spiked into blood and compare this with antibody‐based cell detection. USH1C and CKMT1A are expressed in healthy colon tissue and CRC cell lines, while only low‐level expression can be detected in healthy white blood cells (WBCs). The qPCR reaction shows a near‐perfect amplification efficiency for all primer targets with minimal interference of WBC cDNA. Spike‐in of 10 cancer cells in 3 mL blood can be detected and statistically separated from control blood using the RT‐qPCR assay after rVAR2 capture (p < 0.01 for both primer targets, Mann‐Whitney test). Our results provide a validated workflow for highly sensitive detection of magnetically enriched cancer cells.",
keywords = "Cancer, Circulating tumor cells (CTCs), Detection strategies, Diagnostics, Polymerase chain reaction (PCR), Rare cell detection, RVAR2",
author = "Bang‐christensen, {Sara R.} and Viatcheslav Katerov and J{\o}rgensen, {Amalie M.} and Tobias Gustavsson and Swati Choudhary and Theander, {Thor G.} and Ali Salanti and Allawi, {Hatim T.} and Mette Agerb{\ae}k",
note = "Publisher Copyright: {\textcopyright} 2021 by the authors. Licensee MDPI, Basel, Switzerland.",
year = "2021",
doi = "10.3390/cancers13235881",
language = "English",
volume = "13",
journal = "Cancers",
issn = "2072-6694",
publisher = "M D P I AG",
number = "23",

}

RIS

TY - JOUR

T1 - Detection of VAR2CSA‐captured colorectal cancer cells from blood samples by real‐time reverse transcription PCR

AU - Bang‐christensen, Sara R.

AU - Katerov, Viatcheslav

AU - Jørgensen, Amalie M.

AU - Gustavsson, Tobias

AU - Choudhary, Swati

AU - Theander, Thor G.

AU - Salanti, Ali

AU - Allawi, Hatim T.

AU - Agerbæk, Mette

N1 - Publisher Copyright: © 2021 by the authors. Licensee MDPI, Basel, Switzerland.

PY - 2021

Y1 - 2021

N2 - Analysis of circulating tumor cells (CTCs) from blood samples provides a non‐invasive approach for early cancer detection. However, the rarity of CTCs makes it challenging to establish assays with the required sensitivity and specificity. We combine a highly sensitive CTC capture assay exploiting the cancer cell binding recombinant malaria VAR2CSA protein (rVAR2) with the detection of colon‐related mRNA transcripts (USH1C and CKMT1A). Cancer cell transcripts are detected by RT‐qPCR using proprietary Target Enrichment Long‐probe Quantitative Amplified Signal (TELQAS) technology. We validate each step of the workflow using colorectal cancer (CRC) cell lines spiked into blood and compare this with antibody‐based cell detection. USH1C and CKMT1A are expressed in healthy colon tissue and CRC cell lines, while only low‐level expression can be detected in healthy white blood cells (WBCs). The qPCR reaction shows a near‐perfect amplification efficiency for all primer targets with minimal interference of WBC cDNA. Spike‐in of 10 cancer cells in 3 mL blood can be detected and statistically separated from control blood using the RT‐qPCR assay after rVAR2 capture (p < 0.01 for both primer targets, Mann‐Whitney test). Our results provide a validated workflow for highly sensitive detection of magnetically enriched cancer cells.

AB - Analysis of circulating tumor cells (CTCs) from blood samples provides a non‐invasive approach for early cancer detection. However, the rarity of CTCs makes it challenging to establish assays with the required sensitivity and specificity. We combine a highly sensitive CTC capture assay exploiting the cancer cell binding recombinant malaria VAR2CSA protein (rVAR2) with the detection of colon‐related mRNA transcripts (USH1C and CKMT1A). Cancer cell transcripts are detected by RT‐qPCR using proprietary Target Enrichment Long‐probe Quantitative Amplified Signal (TELQAS) technology. We validate each step of the workflow using colorectal cancer (CRC) cell lines spiked into blood and compare this with antibody‐based cell detection. USH1C and CKMT1A are expressed in healthy colon tissue and CRC cell lines, while only low‐level expression can be detected in healthy white blood cells (WBCs). The qPCR reaction shows a near‐perfect amplification efficiency for all primer targets with minimal interference of WBC cDNA. Spike‐in of 10 cancer cells in 3 mL blood can be detected and statistically separated from control blood using the RT‐qPCR assay after rVAR2 capture (p < 0.01 for both primer targets, Mann‐Whitney test). Our results provide a validated workflow for highly sensitive detection of magnetically enriched cancer cells.

KW - Cancer

KW - Circulating tumor cells (CTCs)

KW - Detection strategies

KW - Diagnostics

KW - Polymerase chain reaction (PCR)

KW - Rare cell detection

KW - RVAR2

U2 - 10.3390/cancers13235881

DO - 10.3390/cancers13235881

M3 - Journal article

C2 - 34884994

AN - SCOPUS:85119608746

VL - 13

JO - Cancers

JF - Cancers

SN - 2072-6694

IS - 23

M1 - 5881

ER -

ID: 285797628