Novel homogenous time-resolved fluorometric RT-PCR assays for quantification of PSA and hK2 mRNAs in blood
Rissanen, Maria ; Helo, Pauliina ; Vaananen, Riina-Minna ; Wahlroos, Veikko ; Lilja, Hans LU
; Nurmi, Martti
; Pettersson, Kim
and Nurmi, Jussi
(2007)
In Clinical Biochemistry
40(1-2).
p.111-118
- Abstract
- Objectives: The purpose of this study was to design, validate, and optimize internally standardized real-time quantitative RT-PCR assays and to identify and avoid problems with assay reliability and examine the impact of an exogenous internal standard. Design and methods: The model system consisted of internally standardized quantitative real-time RT-PCR assays specific for PSA and hK2 mRNA based on time-resolved fluorometric detection of lanthanide chelates. Results: Reproducibility was best when large copy numbers (> 5000 per milliliter blood) were analyzed. Addition of an exogenous target-mimicking internal standard had no significant effect on the reproducibility of the method, but increased the calculated copy numbers by an average... (More)
- Objectives: The purpose of this study was to design, validate, and optimize internally standardized real-time quantitative RT-PCR assays and to identify and avoid problems with assay reliability and examine the impact of an exogenous internal standard. Design and methods: The model system consisted of internally standardized quantitative real-time RT-PCR assays specific for PSA and hK2 mRNA based on time-resolved fluorometric detection of lanthanide chelates. Results: Reproducibility was best when large copy numbers (> 5000 per milliliter blood) were analyzed. Addition of an exogenous target-mimicking internal standard had no significant effect on the reproducibility of the method, but increased the calculated copy numbers by an average of 2-fold. Conclusions: We developed an internally standardized, specific and reproducible real-time RT-PCR analysis method for PSA and hK2 mRNA in circulating cells in the bloodstream. Both PSA and hK2 assays are sufficiently sensitive to detect two LNCaP cells per milliliter whole blood. (c) 2006 The Canadian Society of Clinical Chemists. All rights reserved. (Less)
Please use this url to cite or link to this publication:
https://lup.lub.lu.se/record/676706
- author
- Rissanen, Maria
; Helo, Pauliina
; Vaananen, Riina-Minna
; Wahlroos, Veikko
; Lilja, Hans
LU
; Nurmi, Martti
; Pettersson, Kim
and Nurmi, Jussi
- organization
- publishing date
- 2007
- type
- Contribution to journal
- publication status
- published
- subject
- keywords
- sensitivity and, tissue kallikreins, specificity, reverse transcriptase polymerase chain reaction, neoplasm circulating cells, reproducibility of results
- in
- Clinical Biochemistry
- volume
- 40
- issue
- 1-2
- pages
- 111 - 118
- publisher
- Elsevier
- external identifiers
-
- wos:000243631100019
- scopus:33845764381
- ISSN
- 1873-2933
- DOI
- 10.1016/j.clinbiochem.2006.10.005
- language
- English
- LU publication?
- yes
- id
- c5a54b8c-99ea-47dc-81ed-85464ef1ccd4 (old id 676706)
- date added to LUP
- 2016-04-01 11:33:49
- date last changed
- 2022-01-26 07:03:42
@article{c5a54b8c-99ea-47dc-81ed-85464ef1ccd4,
abstract = {{Objectives: The purpose of this study was to design, validate, and optimize internally standardized real-time quantitative RT-PCR assays and to identify and avoid problems with assay reliability and examine the impact of an exogenous internal standard. Design and methods: The model system consisted of internally standardized quantitative real-time RT-PCR assays specific for PSA and hK2 mRNA based on time-resolved fluorometric detection of lanthanide chelates. Results: Reproducibility was best when large copy numbers (> 5000 per milliliter blood) were analyzed. Addition of an exogenous target-mimicking internal standard had no significant effect on the reproducibility of the method, but increased the calculated copy numbers by an average of 2-fold. Conclusions: We developed an internally standardized, specific and reproducible real-time RT-PCR analysis method for PSA and hK2 mRNA in circulating cells in the bloodstream. Both PSA and hK2 assays are sufficiently sensitive to detect two LNCaP cells per milliliter whole blood. (c) 2006 The Canadian Society of Clinical Chemists. All rights reserved.}},
author = {{Rissanen, Maria and Helo, Pauliina and Vaananen, Riina-Minna and Wahlroos, Veikko and Lilja, Hans and Nurmi, Martti and Pettersson, Kim and Nurmi, Jussi}},
issn = {{1873-2933}},
keywords = {{sensitivity and; tissue kallikreins; specificity; reverse transcriptase polymerase chain reaction; neoplasm circulating cells; reproducibility of results}},
language = {{eng}},
number = {{1-2}},
pages = {{111--118}},
publisher = {{Elsevier}},
series = {{Clinical Biochemistry}},
title = {{Novel homogenous time-resolved fluorometric RT-PCR assays for quantification of PSA and hK2 mRNAs in blood}},
url = {{http://dx.doi.org/10.1016/j.clinbiochem.2006.10.005}},
doi = {{10.1016/j.clinbiochem.2006.10.005}},
volume = {{40}},
year = {{2007}},
}