In the May/June issue of the PDA Journal of Pharmaceutical Science and Technology, Luis Jimenex provides an outstanding overview of genetic sequencing applications as well as a case study in using molecular technologies to rapidly detect microbial contamination, provide a more accurate identification of microorganisms, understand the types of microbes in pharmaceutical laboratories, and make pharmaceutical processes more efficient. The full reference and abstract are provided below.
Molecular Applications to Pharmaceutical Processes and Cleanroom Environments. LUIS JIMENEZ. PDA J Pharm Sci and Tech 2011, 65 242-253.
ABSTRACT:
This review article discusses several technologies using the power of genetic analysis that were used to rapidly detect microbial contamination, provide a more accurate identification of microorganisms, understand the microbial diversity in cleanroom environments, and optimize pharmaceutical processes. Nucleic acids were extracted and purified from enrichment cultures of product suspensions, growing cultures, microbial isolates, and directly from water and cleanroom surfaces. The extracted microbial DNA was either amplified using different polymerase chain reaction (PCR) assays or analyzed using DNA microarray technology. PCR assays were targeting genes coding for structural proteins, ribosomal subunits, or catabolic enzymes. Different PCR formats were tested ranging from single gene amplification to multiplexing. The detection of the amplified fragments was performed by qualitative or quantitative analysis. In some studies the amplified products were cloned/sequenced or sequenced to accurately determine the identity and diversity of the microbes present in the samples. DNA microarray analyses provided a higher multiplexing capability than PCR assays, as more than 3000 genes were analyzed in a single reaction, providing a higher resolution and specificity of the microorganisms present in cleanroom environments and pharmaceutical samples.