Quantitation of estrogen receptor mRNA in breast carcinoma by branched DNA assay.

A quantitative nucleic acid hybridization assay for determination of estrogen receptor (ER) mRNA in breast carcinoma is described. The assay, which is based on the branched DNA (bDNA) technology, requires 20 mg of tissue, is simple, highly specific, and reproducible, and correlates reasonably well with an established methodology (r = 0.87). The assay has a dynamic range of 3 x 10(3)-6 x 10(7) copies of ER mRNA per well. ER message as high as 2.5 x 10(6) copies per well could be detected in normal breast tissues. Thus a sensitivity of 3 x 10(3) ER copies per well was sufficient to analyze clinical specimens. In the present studies, accurate measurement of tissue weight enabled direct reporting of the ER mRNA values as the end point results. The bDNA assay provides a useful tool for the detection and quantitation of ER mRNA in research and routine clinical laboratories.

Quantitation of progesterone receptor mRNA in breast carcinoma by branched DNA assay.

Expression of progesterone receptor (PR) mRNA is indicative of a normal gene regulation mechanism mediated by functional estrogen receptor (ER). A simple assay which can reliably detect and quantitate PR mRNA levels in a small amount of tissue will be of value for studying functional status of ER. We have developed a quantitative nucleic acid hybridization assay for PR mRNA in breast carcinoma. The assay, which is based on the branched DNA (bDNA) technology, is simple, highly specific, and reproducible, requires 20 mg of tissue, and correlates reasonably well (r = 0.86) with an established methodology. The assay has a dynamic range of 3 x 10(3)-6 x 10(7) copies of PR mRNA per well. PR message as high as 3.9 x 10(5) copies per well could be detected in normal breast tissues. Thus a sensitivity of 3 x 10(3) PR copies per well was sufficient for testing clinical samples. In the present studies, accurate measurement of tissue weight enabled direct reporting of the PR mRNA values as the end point results. The bDNA assay provides a useful tool for the detection and quantitation of PR mRNA in research and routine clinical laboratories.

[Correlation between hepatitis C virus (HVC) RNA and anti-HVC antibodies in a hemodialysis population]. / Corrélation entre l'ARN du virus de l'hépatite C (VHC) et les anticorps anti-VHC dans une population d'hémodialysés.

Polymerase chain reaction (PCR) was applied to detect HCV-RNA in 75 hemodialyzed patients. Anti-HCV status was determined by ELISA-2 and by RIBA-2 for reactive samples by ELISA. ALT levels were monthly determined during the year preceding the end of the study. For 60 patients, anti-HCV serology was known since 1989 and 39 of them were tested for the presence of HCV-RNA at least four times during the 2 preceding years. The 9 patients who were negative for anti-HCV antibodies were negative by PCR. Of the 7 patients with an indeterminate profile by RIBA-2, 3 were positive by PCR: 1/1 with C-33c band only and 2/6 with C22-3 band only. Of the 59 patients reactive by RlBA-2, 57 were HCV-RNA positive. Of the 2 HCV-RNA negative patients, one had been PCR positive before interferon therapy. Of the 38 patients without acute hepatitis tested by PCR on 5 successive samples, all the specimens of 11 and 23 patients were HCV-RNA negative and HCV-RNA positive respectively. In 4 patients, a transient viremia was observed. The group of HCV-RNA positive patients had mean ALT levels greater than those who were negative. A correlation was established between HCV infection and both the time on dialysis and the number of blood transfusions. A high concordance (97%) was observed between antibodies to HCV and HCV-RNA.