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Marcel Jacobs

Publications and source records attributed to Marcel Jacobs.

2 recordsLinked to original sources

Multiparametric duplex real-time nucleic acid sequence-based amplification assay for mRNA profiling.

Nucleic acid sequence-based amplification (NASBA) is a sensitive isothermal transcription-based amplification method. We have developed real-time NASBA assays to detect mRNA coding for the estrogen receptor alpha (ESR1) and the progesterone receptor (PGR) in breast tumors by means of duplex reactions using cyclophilin B (PPIB) as the normalizing gene. Both the ESR1/PPIB and PGR/PPIB duplex NASBA assays are highly sensitive, specific, and reproducible. Quantification is determined using external standard calibration curves and the ratio between the number of target and housekeeping gene mRNA copies. Amplification of the target gene in the duplex NASBA assay was disrupted when this latter was mixed with a large amount of the housekeeping PPIB gene, suggesting that it is preferable for the normalizing gene chosen to have an expression level comparable to the target gene. Sensitivity and robustness of the duplex NASBA assays were assessed in breast cancer cell lines. Such a rapid and easy-to-use multiparametric duplex real-time NASBA assay could also advantageously be set up for other mRNA profiling applications.

Biological Assay↗

Rate of human papillomavirus clearance after treatment of cervical intraepithelial neoplasia.

OBJECTIVE: To investigate the rate of clearance of human papillomavirus (HPV) infection after surgical treatment for cervical intraepithelial neoplasia (CIN). METHODS: One hundred nine women with CIN I-III, treated with cryosurgery or conization at a university hospital, were observed with cervical HPV deoxyribonucleic acid (DNA) testing by general primer polymerase chain reaction and HPV typing at 0, 3, 6, 9, 12, and 24 months after treatment. Penile HPV DNA was analyzed from current sexual partners. RESULTS: Eighty-one percent of evaluable women were HPV DNA positive at treatment or enrollment. One year later, seven women (9%) remained positive for the same HPV type. Most women had cleared the HPV infection diagnosed at treatment within 3 months. The cryotherapy group had lower CIN grades, was younger, and had a slower HPV clearance rate (P <.002). Only four couples had HPV DNA of the same type detected. CONCLUSION: Surgical treatment of CIN usually results in clearance of HPV infection within 3 months. Human papillomavirus DNA testing may be useful as a rapid intermediate end point for monitoring the efficacy of treatments.

Adult↗