High throughput testing for presence of microorganisms in a biological sample

A method for determining whether two or more species of specified microorganisms are present within a biological sample that potentially comprises the microorganisms comprising:

• (a) immobilizing the biological sample on and/or within a solid substrate at a first location;
• (b) transferring the immobilized biological sample to at least a remote second location and performing an extraction step on the solid substrate so as to extract any microorganism DNA immobilized on and/or within the solid substrate;
• (c) performing a nucleic acid amplification step on microorganism DNA extracted in step (b), wherein the amplification step comprises a plurality of amplification primers that are directed towards amplification of a plurality of highly conserved sequences from the two or more specific microorganisms, and wherein amplified sequences are designated as target sequences;
• and wherein a plurality of control competitor nucleic acid sequences are combined with the target sequences prior to the nucleic acid amplification step of part (c), wherein each competitor nucleic acid sequence is identical to a corresponding target sequence except that the competitor nucleic acid sequence comprises a sequence variation at a specified position compared to the corresponding target sequence;
• (d) combining the target sequences with a plurality of primer sequences comprised within a diagnostic multiplexing panel (DMP), wherein each primer sequence facilitates genotyping of the target sequence;
• (e) performing a primer extension reaction on the combination of target sequences and the DMP present in (d), thereby producing a DMP reaction product;
• and
• (f) analysing the DMP reaction product so as to determine genotype of any target sequences that are present and correlating the genotype of the target sequences in the reaction product with the identification of specified microorganisms present in the biological sample;
• and wherein said specified
• microorganisms are selected from one or more of the group consisting of bacteria, fungi, viruses and protozoa.