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Laboratory techniques can enhance
traditional outbreak investigation. Tests have been developed that
go beyond species identification and give information about the relatedness
of microorganisms. Strain typing is most often performed on the bacterial
species responsible for nosocomial infections, but mycobacteria, yeast,
filamentous fungi and viruses may also by typed. Such strain typing
systems are based on the fact that related microbial isolates are
derived from a single clone and share characteristics which differ
from unrelated isolates.
Clones are defined as indistinguishable isolates, derived from the
same parent strain. Subsequent generations of organisms often begin
to diverge from the parent. As divergence occurs progeny strains
will share most characteristics with the parent, but differences
will appear. These progeny strains may be termed clonally-related
to the parent. It follows that isolates which are clones or clonally-related
may be part of an outbreak. Thus, strain typing helps the epidemiologist
determine when to initiate additional investigation. Related organisms
may be derived from a point source such as a contaminated solution,
fomite or colonized health care worker. They could also be transmitted
from patient-to-patient via health care worker hands or medical
equipment. It is easy to understand the role that culture and characterization
of microbes, including those from non-patient sources, may play
in helping the infection control team plan appropriate interventions
in outbreak settings. On the other hand, when organisms are shown
to be unrelated, an outbreak of a single strain type may be ruled-out.
In addition to outbreak investigation, strain typing can help the
epidemiologist track a particular strain within a given institution
and through the wider health care community. Clinicians have also
applied these techniques to assess whether multiple infections with
the same species over time represents persistent infection or acquisition
of a new strain. Determining whether multiple positive blood cultures
are caused by the same organism or several different strains of
a skin contaminant is another clinical use for these methods. A
number of different laboratory methods have been developed for strain
delineation. Simple phenotypic tests such as biochemical profiling,
serotyping and antibiotic susceptibility patterns characterize bacteria
based on the expression of genes and are an important first screen
of relatedness. However, many organisms share phenotypic traits,
yet are genetically distinct. Susceptibility of a bacterium to a
panel of antibiotics (the antibiogram) is probably the most useful
of this class of tests because these data are readily available
at the time of organism identification. If isolates have different
antibiotic susceptibilities, they are probably not closely related.
Many outbreaks, however, are caused by strains with similar antibiograms
such as the highly resistant methicillin resistant Staphylococcus
aureus (MRSA) and vancomycin resistant Enterococcus. The limited
variability in susceptibility patterns for such strains necessitates
the use of a molecular method. Genotypic or molecular methods are
based on differences in the presence, size and sequence of genetic
material. A number of techniques have been developed to analyze
and compare the chromosome or plasmid content of strains. Most of
these tests have been designed to yield fragments of DNA of variable
lengths which are then separated by gel electrophoresis to give
an array of bands called the "DNA fingerprint." The techniques used
most commonly for nosocomial pathogens include: pulsed-field gel
electrophoresis (PFGE), restriction fragment length polymorphism
(RFLP), Ribotyping, and PCR amplification of random (RAPD) or extra-genic sequences.
The first three techniques are examples of methods used in our laboratory.
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