ACTIVITY NO.

2 (MOLECULAR BASIS OF DETECTION)

1. What are the traditional basis of detection and identification of pathogenic

microorganisms. Explain each.
Traditional approaches for microbial detection and identification include microbial
cultivation, immunological (e.g., antibody-based) assays, and nucleic acid detection
schemes—especially amplification methods such as PCR.
Microbial cultivation can be used to generate pure cultures, which provide a continuous
supply of cells from the same species or strain. Such cultures can be used to
investigate microbial traits in experiments performed in replicates, which improves
reproducibility and statistical confidence.
Immunological Assays is a test that uses the binding of antibodies to antigens to identify
and measure certain substances. Immunoassays may be used to diagnose disease.
Also, test results can provide information about a disease that may help in planning
treatment.
Nucleic acid detection is a key technique that enables the detection of specific
nucleotide sequences and ultimately unveils the presence of a pathogen or early onset
of a diseased state. Over the last few years, improvements in nucleic-acid based
diagnostics as in polymerase chain reaction (PCR) and sequence alignment studies
have transformed the way clinical laboratories now diagnose pathogens, thereby
allowing rapid and effective response to the treatment of infections at an early stage.

2. What are the molecular basis of detection and identification of pathogenic

microorganisms. Explain each.
Single polymerase chain reaction
This is a polymerase chain reaction (PCR) involving the use of a single primer set
(which targets a specific gene) to detect an organism. The primer set can be designed
for specific species and can detect the target organism in the presence of others.
Multiplex polymerase chain reaction
This is a modification of polymerase chain reaction that uses multiple primers within a
single PCR mixture to detect, identify and/or differentiate bacteria.
Other polymerase chain reaction assays
These encompass commonly available modified PCR techniques that are yet to be
applied to foodborne pathogens isolated from ducks. They include real-time PCR,
nested PCR and reverse-transcription PCR.
 3. Compare the traditional and molecular basis of detection and identification
of pathogenic microorganisms.
In contrast to phenotypic testing, molecular testing uses the microorganism's genome,
transcriptome, or proteome as an analyte. Viruses can have DNA or RNA genomes,
whereas bacteria, fungi, and parasites have DNA genomes. Prions are proteins that
cause transmissible encephalopathies such as Creutzfeldt–Jakob disease. DNA, RNA,
and protein characterization was developed to detect and identify novel species as well
as to further characterize and classify existing creatures, such as the influenza virus.