Detection of critical infections associated with pregnancy and birth such as intraamniotic infection and neonatal sepsis via commercially available laboratory culture techniques can be difficult because:

• The fastidious nature of many bacteria commonly associated with these infections such as Mycoplasma sp., can make them difficult to culture. The specialized techniques required to reliably cultivate these organisms are not routinely used in clinical settings.

• The specimen of biological fluid obtained for diagnostic testing may contain an insufficient infectious inoculum to be detected via culture.

• Previous antibiotic use can result in the inability of culture to detect pathogenic organisms.

Molecular microbiology techniques such as Quantitative/Real-Time Polymerase Chain Reaction (qPCR) and Pyrosequencing can detect microbes independently of culture. These techniques amplify highly conserved gene sequences that allow detection and broad taxonomic identification of pathogenic microorganisms in biological fluids.

Ribosomal RNA is the most conserved and least variable gene in all cells including bacteria. For this reason, genes that encode ribosomal RNA have been used extensively to identify medically important microorganisms such as bacteria and fungi. The 16S and 23S rRNA gene sequences are typically used to identify bacteria species while the corresponding 18S rRNA gene sequences are used to identify fungi.

ProteoGenix nucleic acid-based molecular diagnostics technologies will be based on quantitative polymerase chain reaction (qPCR) amplification of ribosomal DNA from both bacteria and fungi, as well as other unique microbial gene sequences.. ProteoGenix also has developed proprietary specimen collection, transport and preparation systems that uniquely optimize the detection and identification of pathogenic bacteria and fungi associated with critical maternal, fetal and neonatal disorders.

The unique, identifying DNA sequences characteristic of individual microorganisms are commonly identified in the laboratory by hybridizing an amplified specific microbial gene sequence to a specific fluorescent molecular probe that can be used to detected and quantified the microorganism present in the clinical sample.

In addition to this test approach, ProteoGenix uses breakthrough Pyrosequencing technology to quickly, accurately and specifically identify a host of potential pathogenic microbes down to the species level. Pyrosequencing allows for this simultaneous identification of multiple microbial gene sequences using DNA sequencing technology consisting of an enzyme-cascade system of four enzymes and specific substrates. Also known as “sequencing by synthesis”, Pyrosequencing produces light in the reaction whenever a nucleotide is incorporated as a base pair with the complementary base in a DNA template strand. Pyrosequencing is a rapid process allowing the sequencing of 30 to 40 base pairs in less than an hour.

Nucleic acid-based molecular diagnostic tests are accurate tools for detection and identification of pathogenic microorganisms. These techniques can be used as diagnostic tools themselves for detection of infection and also to aid in the selection of proteomic biomarkers that specifically correlate with critical conditions of pregnancy.