Detection of micrometastasis by cytokeratin 20 RT-PCR is limited due to stable background transcription in granulocytes.
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Detection of micrometastasis by cytokeratin 20 RT-PCR is limited due to stable background transcription in granulocytes. / Jung, Roman; Petersen, K; Krüger, W; Wolf, M; Wagener, C; Zander, A; Neumaier, M.
In: BRIT J CANCER, Vol. 81, No. 5, 5, 1999, p. 870-873.Research output: SCORING: Contribution to journal › SCORING: Journal article › Research › peer-review
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T1 - Detection of micrometastasis by cytokeratin 20 RT-PCR is limited due to stable background transcription in granulocytes.
AU - Jung, Roman
AU - Petersen, K
AU - Krüger, W
AU - Wolf, M
AU - Wagener, C
AU - Zander, A
AU - Neumaier, M
PY - 1999
Y1 - 1999
N2 - The reverse transcription polymerase chain reaction (RT-PCR) amplification of cytokeratin 20 (CK20) mRNA is considered a promising candidate method for the detection of circulating tumour cells in bone marrow and peripheral blood of cancer patients. In this study we have investigated the diagnostic specificity of the CK20 mRNA detection in samples from healthy donors (HD; n = 33), intensive care units patients (ICU; n = 20) and bone marrow obtained from patients suffering from chronic inflammatory diseases (CID; n = 14). RNAs purified from stabilized lysates showed positive results in 24% of the HD group (8/33), 35% of the ICU group (8/20) and in 40% of the CID group (5/14). The use of Ficoll gradients to separate nucleated cells completely restored the specificity of this CK20 RT-PCR assay. The CK20-expressing cells are positively identified to belong to the granulocyte fraction of leucocytes, which appear to express the gene on a background level. Our results demonstrate for the first time that CK20 mRNA expression is not limited to epithelium. Its occurrence in normal granulocytes has to be considered in tests designed to detect circulating cancer cells or micrometastases.
AB - The reverse transcription polymerase chain reaction (RT-PCR) amplification of cytokeratin 20 (CK20) mRNA is considered a promising candidate method for the detection of circulating tumour cells in bone marrow and peripheral blood of cancer patients. In this study we have investigated the diagnostic specificity of the CK20 mRNA detection in samples from healthy donors (HD; n = 33), intensive care units patients (ICU; n = 20) and bone marrow obtained from patients suffering from chronic inflammatory diseases (CID; n = 14). RNAs purified from stabilized lysates showed positive results in 24% of the HD group (8/33), 35% of the ICU group (8/20) and in 40% of the CID group (5/14). The use of Ficoll gradients to separate nucleated cells completely restored the specificity of this CK20 RT-PCR assay. The CK20-expressing cells are positively identified to belong to the granulocyte fraction of leucocytes, which appear to express the gene on a background level. Our results demonstrate for the first time that CK20 mRNA expression is not limited to epithelium. Its occurrence in normal granulocytes has to be considered in tests designed to detect circulating cancer cells or micrometastases.
M3 - SCORING: Zeitschriftenaufsatz
VL - 81
SP - 870
EP - 873
JO - BRIT J CANCER
JF - BRIT J CANCER
SN - 0007-0920
IS - 5
M1 - 5
ER -