Tumour diagnosis by PCR-based detection of tumour cells.
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Tumour diagnosis by PCR-based detection of tumour cells. / Nollau, P; Jung, Roman; Neumaier, M; Wagener, C.
in: Scand J Clin Lab Invest Suppl, Jahrgang 221, 1995, S. 116-121.Publikationen: SCORING: Beitrag in Fachzeitschrift/Zeitung › SCORING: Zeitschriftenaufsatz › Forschung › Begutachtung
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TY - JOUR
T1 - Tumour diagnosis by PCR-based detection of tumour cells.
AU - Nollau, P
AU - Jung, Roman
AU - Neumaier, M
AU - Wagener, C
PY - 1995
Y1 - 1995
N2 - Tumour cells shed from solid primary tumours can be detected by the polymerase chain reaction (PCR) based on the selective amplification of mutated tumour genes or of genes expressed in a tissue specific manner. When tumour specific alterations are amplified, few tumour cells can be detected in excess of normal cells derived from the same tissue. Thus, malignant cells can be detected specifically in pancreatic juice, stool, urine, and sputum. Here we describe the adaptation of the mutant enriched PCR in conjunction with the introduction of artificial primer mediated restriction sites to the selective amplification of mutant K-ras genes in stool samples from patients with colorectal carcinomas. In reconstitution experiments, down to 10 colorectal carcinoma cells could be detected in 100 mg of stool. For the diagnosis of micrometastatic disease, a sensitive and specific technique was established based on the reverse transcription of mRNA specific for the carcinoembryonic antigen followed by the amplification of the cDNA (RT-PCR). Attempts to establish a specific RT-PCR for cytokeratin-18 failed because of the existence of at least one processed pseudogene.
AB - Tumour cells shed from solid primary tumours can be detected by the polymerase chain reaction (PCR) based on the selective amplification of mutated tumour genes or of genes expressed in a tissue specific manner. When tumour specific alterations are amplified, few tumour cells can be detected in excess of normal cells derived from the same tissue. Thus, malignant cells can be detected specifically in pancreatic juice, stool, urine, and sputum. Here we describe the adaptation of the mutant enriched PCR in conjunction with the introduction of artificial primer mediated restriction sites to the selective amplification of mutant K-ras genes in stool samples from patients with colorectal carcinomas. In reconstitution experiments, down to 10 colorectal carcinoma cells could be detected in 100 mg of stool. For the diagnosis of micrometastatic disease, a sensitive and specific technique was established based on the reverse transcription of mRNA specific for the carcinoembryonic antigen followed by the amplification of the cDNA (RT-PCR). Attempts to establish a specific RT-PCR for cytokeratin-18 failed because of the existence of at least one processed pseudogene.
M3 - SCORING: Zeitschriftenaufsatz
VL - 221
SP - 116
EP - 121
ER -
