Competitive RT-PCR to quantify CFTR mRNA in human endometrium

Because the down-regulation by progesterone of cystic fibrosis transmembrane conductance regulator (CFTR) expression could be a useful specific marker to define the state of implant receptivity in endometrium, a competitive reverse transcription-polymerase chain reaction (RT-PCR) was developed for q...

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Bibliographic Details
Published in:Clinical chemistry (Baltimore, Md.) Md.), 1996-11, Vol.42 (11), p.1765-1769
Main Authors: Mularoni, A, Adessi, GL, Arbez-Gindre, F, Agnani, G, Nicollier, M
Format: Article
Language:English
Subjects:
Binding, Competitive
Biological and medical sciences
Coloring Agents
Cystic Fibrosis Transmembrane Conductance Regulator - genetics
DNA Primers
Electrophoresis, Polyacrylamide Gel
Endometriosis - metabolism
Endometrium - chemistry
Endometrium - pathology
Ethidium
Female
Genital system. Mammary gland
Humans
Hyperplasia
Investigative techniques, diagnostic techniques (general aspects)
Medical sciences
Menstrual Cycle
Nucleic Acid Heteroduplexes
Pathology. Cytology. Biochemistry. Spectrometry. Miscellaneous investigative techniques
Polymerase Chain Reaction
RNA, Messenger - analysis
RNA-Directed DNA Polymerase
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Summary:Because the down-regulation by progesterone of cystic fibrosis transmembrane conductance regulator (CFTR) expression could be a useful specific marker to define the state of implant receptivity in endometrium, a competitive reverse transcription-polymerase chain reaction (RT-PCR) was developed for quantifying the CFTR mRNA concentration in human endometrial samples. A competitor RNA was constructed with the same sequence as the CFTR sequence except for a 20-nucleotide insertion in the middle. The amplified products were separated by polyacrylamide gel electrophoresis. The ratio of CFTR band areas to competitor band areas provided the basis of quantification. Using this competitive RT-PCR, we measured CFTR mRNA in human endometrial samples taken at different periods of the menstrual cycle, in endometriosis, and in hyperplasia. Results show that the method is suitable for measuring the concentration of CFTR mRNA.
ISSN:0009-9147
1530-8561
DOI:10.1093/clinchem/42.11.1765