Format

Send to

Choose Destination
See comment in PubMed Commons below
J Cancer Res Clin Oncol. 2014 Jan;140(1):145-50. doi: 10.1007/s00432-013-1555-5. Epub 2013 Nov 27.

Digital PCR quantification of miRNAs in sputum for diagnosis of lung cancer.

Author information

1
Department of Pathology, The University of Maryland Greenebaum Cancer Center, University of Maryland School of Medicine, 10 South Pine Street, MSTF 7th Floor, Baltimore, MD, 21201-1192, USA.

Abstract

PURPOSE:

MicroRNAs (miRNAs) play important roles in the initiation and progression of lung cancer. Measuring miRNA expression levels in sputum could provide a potential approach for the diagnosis of lung cancer. The emerging digital PCR is a straightforward technique for precise, direct, and absolute quantification of nucleic acids. The objective of the study was to investigate whether digital PCR could be used to quantify miRNAs in sputum for lung cancer diagnosis.

METHODS:

We first determined and compared dynamic ranges of digital PCR and conventional quantitative reverse transcriptase PCR (qRT-PCR) for miRNA quantification using RNA isolated from sputum of five healthy individuals. We then used digital PCR to quantify copy number of two lung cancer-associated miRNAs (miR-31 and miR-210) in 35 lung cancer patients and 40 cancer-free controls.

RESULTS:

Copy number of the miRNAs measured by digital PCR displayed a linear response to input cDNA amount in a twofold dilution series over seven orders of magnitude. miRNA quantification determined by digital PCR assay was in good agreement with that obtained from qRT-PCR analysis in sputum. Furthermore, combined quantification of miR-31 and miR-210 copy number by using digital PCR in sputum of the cases and controls provided 65.71 % sensitivity and 85.00 % specificity for lung cancer diagnosis.

CONCLUSION:

As digital PCR becomes more established, it would be a robust tool for quantitative assessment of miRNA copy number in sputum for lung cancer diagnosis.

PMID:
24281335
PMCID:
PMC3898839
DOI:
10.1007/s00432-013-1555-5
[Indexed for MEDLINE]
Free PMC Article
PubMed Commons home

PubMed Commons

0 comments
How to join PubMed Commons

    Supplemental Content

    Full text links

    Icon for Springer Icon for PubMed Central
    Loading ...
    Support Center