A method for simultaneously detecting and quantifying a microbial nucleic acid in a biological sample, said method comprising: a) isolating and purifying said microbial nucleic acid b) providing a reaction mixture comprising a first and a second control nucleic acid in different concentrations wherein the first control nucleic acid is a quantitative standard nucleic acid present in a concentration of 20-5000 times the limit of detection of said microbial nucleic acid, and the second control nucleic acid is a qualitative internal control nucleic acid present in a concentration of 1-10 times the limit of detection of said microbial nucleic acid, one or more primer pairs specifically hybridizing to distinct sequence portions of said microbial nucleic acid and to distinct sequence portions of said control nucleic acids, and probes specifically hybridizing to each of the sequences amplified by said one or more primer pairs, wherein said microbial nucleic acid and said first control nucleic acid and said second control nucleic acid hybridize to different probes carrying different labels c) adding the isolated and purified microbial nucleic acid to said reaction mixture d) performing one or more cycling steps, wherein a cycling step comprises an amplifying step, said amplifying step comprising producing one or more amplification products derived from said microbial nucleic acid if present in said sample and producing an amplification product derived from said first control nucleic acid and said second control nucleic acid, and wherein a cycling step comprises a hybridizing step, said hybridizing step comprising hybridizing the sequences amplified by said primer pair with said probes, wherein the probes are labeled with a donor fluorescent moiety and a corresponding acceptor fluorescent moiety and each of the probes carries a different fluorescent dye e) detecting and measuring fluorescent signals generated by the amplification products of said first control nucleic acid and said microbial nucleic acid and being proportional to their concentration, and simultaneously detecting fluorescent signals generated by said amplification product of said second control nucleic acid, wherein the presence of an amplification product of said second control nucleic acid is indicative of an amplification occurring in the reaction mixture even in the absence of an amplification product for said microbial nucleic acid, and determining the quantity of said microbial nucleic acid in said biological sample by comparison of the signals generated by said microbial nucleic acid and said first control nucleic acid. a) isolating and purifying said microbial nucleic acid b) providing a reaction mixture comprising a first and a second control nucleic acid in different concentrations wherein the first control nucleic acid is a quantitative standard nucleic acid present in a concentration of 20-5000 times the limit of detection of said microbial nucleic acid, and the second control nucleic acid is a qualitative internal control nucleic acid present in a concentration of 1-10 times the limit of detection of said microbial nucleic acid, one or more primer pairs specifically hybridizing to distinct sequence portions of said microbial nucleic acid and to distinct sequence portions of said control nucleic acids, and probes specifically hybridizing to each of the sequences amplified by said one or more primer pairs, wherein said microbial nucleic acid and said first control nucleic acid and said second control nucleic acid hybridize to different probes carrying different labels c) adding the isolated and purified microbial nucleic acid to said reaction mixture d) performing one or more cycling steps, wherein a cycling step comprises an amplifying step, said amplifying step comprising producing one or more amplification products derived from said microbial nucleic acid if present in said sample and producing an amplification product derived from said first control nucleic acid and said second control nucleic acid, and wherein a cycling step comprises a hybridizing step, said hybridizing step comprising hybridizing the sequences amplified by said primer pair with said probes, wherein the probes are labeled with a donor fluorescent moiety and a corresponding acceptor fluorescent moiety and each of the probes carries a different fluorescent dye e) detecting and measuring fluorescent signals generated by the amplification products of said first control nucleic acid and said microbial nucleic acid and being proportional to their concentration, and simultaneously detecting fluorescent signals generated by said amplification product of said second control nucleic acid, wherein the presence of an amplification product of said second control nucleic acid is indicative of an amplification occurring in the reaction mixture even in the absence of an amplification product for said microbial nucleic acid, and determining the quantity of said microbial nucleic acid in said biological sample by comparison of the signals generated by said microbial nucleic acid and said first control nucleic acid.