Microbiology is the scientific study of micro-organisms and includes many sub disciplines like bacteriology, virology, mycology, and parasitology: all characterized by the study of organisms too small to be seen by the naked eye. This defining aspect has determined the focus of microbial research over the last century: the need to selectively concentrate or amplify micro-organisms or their components to detect their presence. Earlier efforts into microbial detection involved microscopy, where light was transmitted through or reflected from the sample through lenses to magnify the view of the sample and determine the presence of micro-organisms. The limitations of light microscopy at the time prevented the study of living micro-organisms and it wasn’t until Pasteur (1822-1895) developed techniques to disprove the theory of spontaneous generation, and Robert Koch (1843-1910) developed his ‘postulates’ proving that diseases were caused by specific bacteria, was the era of microbial culture born.

Over the last decade, interest in rapid microbiological methods (RMMs) in the pharmaceutical sector has grown considerably. Technologies such as ATP bioluminescence, solid phase laser cytometry and genetic-based identification systems are being vigorously investigated. Validation and regulatory requirements for such new technologies are beginning to emerge. However, there is a lot of confusion and considerable hesitancy associated with the introduction of these methods into the pharmaceutical sector. The aim of this article is to help clear up some of these issues in light of recent published literature.

Subsequent to the publication of the PDA technical Report Number 33 in 2000 there has been a multitude of articles published in pharmaceutical reviews and commentaries with regard to rapid microbial methods (RMM). The majority of these articles cover, to a greater or lesser degree, the technologies available and summaries of the requirements for validation, implementation and regulatory acceptance. There have also been some articles that have explored the potential reasons for the relatively slow uptake of alternative microbiological testing methods. While these articles address valid issues, few explore in detail the further possibility for the slow uptake; the appropriateness of the currently available RMM technologies.

Arguably microbiology is the oldest of the applied sciences, although early exponents doubtless had no understanding of how the fruits of their labour in fermentation for example, came about. The true forerunners of microbiology as it is recognised today would be Koch, Pasteur, Petri et al, who developed much of our basic understanding of the subject and many of the methods that are still in use approximately 150 years later. In that intervening period our knowledge of human and microbial genetics has dramatically increased and our exploitation of that knowledge has led to the current explosive growth of Biotechnology, such that clinically important biopharmaceuticals can now be produced by insect, plant, bacterial or mammalian cell lines or indeed in the milk of bovines.

In order to meet the challenges demanded by the requirements of Process Analytical Technology (PAT), the modern microbiological laboratory needs to become more innovative in microbial detection, identification and enumeration. Technology is becoming available that will speed up microbiological analysis, potentially allowing pharmaceutical microbiology tests to get as close as is possible to the concepts of PAT. Following on from the article by Bob Johnson1, this article explores the future technologies in greater detail.

The modern microbiological laboratory needs to change and become more innovative in its approach to microbial detection, enumeration and identification.

The purpose of this article is to assess the potential significance of Viable but Non Culturable (VBNC) microorganisms in the pharmaceutical industry; consideration is given to the definition of the VBNC state, current methods for the detection of such organisms are outlined and potentially significant new methods, which may impact the industry, are considered. The article will also discuss the future significance of VBNC organisms within pharmaceutical microbiology and its ongoing development.

The identification of microorganisms from the pharmaceutical production environment has gained an ever greater importance in modern times. Thus the new Aseptic Processing Guide of the FDA recommends the identification of detected isolates from the critical clean room area (grade A or ISO 5) down to the species level and recommends the identification of isolates from the surrounding lesser controlled clean room area (i.e. grade B or ISO 7) down to at least the genus level.

Following on from our latest report in Issue 4 in which we outlined independent views on ‘solutions for the next stage’ in Rapid Microbiological Methods (RMM) from some of the industry’s leading names, Tony Cundell follows up with his own thoughts on this exciting field.

Rapid microbiology is an exciting field for the development of new technologies and applications. There are significant challenges to be overcome and in doing so, great prospects for microbiologists and the companies that provide cutting-edge equipment.

Since the 2000 publication of the PDA Technical Report Number 33 Testing Methods, rapid microbial methods (RMM) have not lived up to their promise for changing routine Quality Control microbiological testing in the pharmaceutical industry. With the next generation of rapid microbial methods emerging from the R&D laboratories of instrument manufacturers, this article will discuss the obstacles to the successful implementation of the currently marketed technologies and strategies to overcome these obstacles.

Timely results for microbial bioburden monitoring of in-process cell culture samples are critical to the production process for recombinant proteins and other biopharmaceutical products.

In the last decade interest in Rapid Microbiological Methods (RMMs) has grown considerably. Technologies such as ATP bioluminescence, solid phase laser cytometry and genetic-based identification systems are being vigorously investigated. Validation and regulatory requirements for such new technologies are beginning to emerge. However, there is a lot of confusion and considerable hesitancy associated with the introduction of these methods into the pharmaceutical sector.