Domingo, Nica A.

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Sakhno NG (2016) Microbial Identification Methods in Pharmaceutical Analysis: Comparison and

Evaluation M J Pharma 1(1): 001

In the article, the author argues that rapid and accurate identification of microorganisms is an essential
part of pharmaceutical analysis. They gave different phenotypic, genotypic and proteotypic methods for
the identification of microorganisms and gave emphasis on the features of each method and factors
affecting the result of identification.

According to the article identification is accomplished by matching characteristics (genotypic or

phenotypic) to an established standard (reference) organism such as a type strain. In phenotypic methods
the microbiologist is able identify microorganisms to the genus and sometimes to the species level based
on a relatively small number of observations and tests which includes culture cultivation and biochemical
assays based on physiological reactions, immunological methods, and fatty acid composition analysis.
Genotypic methods includes nucleic acid amplification techniques and genetic fingerprinting (ribotyping)
which yields a more specific identification of the microorganism. The third group of method which is
referred to as proteotypic methods include Fourier Transform Infrared (FTIR) spectroscopy, and Matrix
Assisted Laser Desorption Ionization-Time of Flight mass spectrometry (MALDI-TOF MS) both are not
discussed in detail.

Because, according to the article, misidentification could lead to inappropriate corrective and preventive
actions and product disposition the authors gave factors affecting the identification for each type of
method mentioned. The limitation of the phenotypic method is that microorganisms are capable of
suddenly altering their phenotypic expressions such as cell size and shape, cellular composition,
biochemical reactivity, antimicrobial susceptibility, etc., due to environmental changes or genetic
mutations. Other limitations given to this method are the relatively large number of cells in pure,
monoclonal culture, and the crucial step of this method, the Gram reaction. Wrong characterization, posed
by gram-variable organisms, assigned to the isolate will lead to the use of wrong microbial testing kit,
resulting to an incorrect result. Genotypic methods, on the other hand, are less dependent on culture
conditions and are more reliable because nucleic acid sequences are highly conserved by microbial
species. Proteotypic methods are also free from restrictions related to conditions of microbial cultivation.
However, these technique are more challenging for the microbiologist and requires more expensive
equipment and supplies.
For the comparison of the different identification systems, the article provided a table that shows the size
of the database, containing the number and types of genera and species identified, for each of the
methods, and the time required to identify the microorganism. (MALDI-TOF MS) and (FTIR) being the
fastest identification method, providing the identity of the microorganism within minutes.

The article stated that fungi is a problematic group of microorganisms which are classified and identified
by the morphological features rather than nutritional and biochemical differences, complicated by the
existence of teleomorph and anamorphic form of the same fungus. Providing different methods to address
the problem such as crystallographic method for the identification of Candida spp. In 16-18 hour culture,
nucleic acid sequencing technique, cultural technique for yeast identification, microscopic features for the
characterization of moulds, and MicroSeq system that is accurate and useful for the identification of
filamentous fungi.

On the conclusion, the authors stated that phenotypic methods based on carbon utilization and
biochemical reactions are still the most widely used pharmaceutical analysis they have supported this
statement by giving consideration to the range of factors starting from the safety of the procedures to the
potential cost savings.

Overall, the article can be related to other application other than the pharmaceutical analysis. Another
field is the hospital or clinical setting where unknown isolates are usually found in normally sterile site
such as blood. The identity of the isolated microorganism is very crucial to the diagnosis and treatment of
the patient.