Document Detail

Improved real-time rt-PCR assays of two colorectal cancer peripheral blood mRNA biomarkers: a pilot study.
MedLine Citation:
PMID:  23279830     Owner:  NLM     Status:  MEDLINE    
BACKGROUND: Efficient screening for detection of colorectal cancer (CRC) at earlier stages reduces its mortality. The purpose of this study was to investigate expression of carcinoembryonic antigen (CEA) and human telomerase reverse transcriptase (hTERT) mRNA in peripheral blood of CRC patients and to present strategies for early detection screen test.
METHODS: Twenty seven patients in non-metastatic stage and 27 healthy individuals were studied. Expression of CEA, hTERT mRNA and 18srRNA (18s subunit of ribosomal RNA, as reference gene) were determined based on real-time RT-PCR on 3 µg of total RNA from blood in 3 separate vials (1 µg per vial).
RESULTS: Positive expression rate of CEA mRNA (78%) and hTERT mRNA (81%) were higher in patient group (P<0.001). These rates were meaningfully higher than the results of individual vials containing only 1 µg of total RNA. Difference between Ct values of markers with 18srRNA ΔCt) was higher in healthy group than patient one. Therefore, a ΔCt cut-off value was determined for distinguishing between true- and false-positive results. Concurrent expression of both markers was found in 67% of the patients, which was higher than healthy cases (11%). Combination of concurrent marker expression with cut-off point strategy increased specificity to 100%.
CONCLUSION: These results showed that concurrent evaluation of marker expression and performing the test on 3 µg of samples in 3 separate vials may increase specificity and sensitivity of real-time RT-PCR for early detection of non-metastatic CRC. However, more investigations with larger numbers of samples are needed to verify these results.
Reza Yadegarazari; Taghi Hassanzadeh; Amir Majlesi; Amir Keshvari; Alireza Monsef Esfahani; Amirsasan Tootoonchi; Nooshin Shabab; Massoud Saidijam
Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't    
Journal Detail:
Title:  Iranian biomedical journal     Volume:  17     ISSN:  2008-823X     ISO Abbreviation:  Iran. Biomed. J.     Publication Date:  2013  
Date Detail:
Created Date:  2013-01-02     Completed Date:  2013-06-25     Revised Date:  2013-07-11    
Medline Journal Info:
Nlm Unique ID:  9814853     Medline TA:  Iran Biomed J     Country:  Iran    
Other Details:
Languages:  eng     Pagination:  15-21     Citation Subset:  IM    
Research Center for Molecular Medicine, Hamadan University of Medical Sciences, Hamadan, Iran.
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MeSH Terms
Aged, 80 and over
Carcinoembryonic Antigen / blood*,  genetics
Colorectal Neoplasms / blood,  diagnosis*,  genetics
Early Detection of Cancer
Middle Aged
Pilot Projects
RNA, Messenger / blood*
RNA, Ribosomal, 18S / blood*
Reverse Transcriptase Polymerase Chain Reaction
Telomerase / blood*,  genetics
Tumor Markers, Biological / blood
Reg. No./Substance:
0/Carcinoembryonic Antigen; 0/RNA, Messenger; 0/RNA, Ribosomal, 18S; 0/Tumor Markers, Biological; EC protein, human; EC

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