Beruflich Dokumente
Kultur Dokumente
( 81)
Supervised by
Prof. Dr. Samir Ahmed Khairallah Professor of Medical Microbiology And Immunology, Department of Medical Microbiology and Immunology, Faculty of Medicine, Mansoura University. Dr. Hamdia Yehia Askar Lecturer of Medical Microbiology And Immunology, Department of Medical Microbiology and Immunology, Faculty of Medicine, Mansoura University.
The
National
Nosocomial
Infections
Surveillance
condition
System
(NNIS)
system)
denes
For most bacterial nosocomial infections, this means that the infection usually becomes evident 48 hours (i.e., the typical incubation period) or more after admission. However, because the incubation period varies with the type of pathogen and to some extent with the patients underlying condition, each infection
NI may be
Nosocomial outbreaks
An outbreak may be defined as
a relatively rapid increase in the incidence of infection or colonization by a certain bacterial strain,
Just
like
endemic
hospital-acquired
Socio-economic impact
Socio-economic impacts of nosocomial infection
specific body sites including blood, respiratory tract or surgical wound; and infections with multidrug resistant micro-
organisms
including
methicillin-resistant
extended-spectrum and
Staphylococcus
lactamase
aureus,
producing
Enterobacteriaceae,
Pseudomonas aeruginosa).
Severity
of
underlying
disease
classification was evaluated at the time of NI occurrence, using an established modified risk
stratification regimen, as
rapidly fatal
ultimately fatal
non-fatal
antibiotics when death occurred and no other obvious causes of death were evident.
For instance, even after proper hand washing and glove usage, MRSA could still be found on the hands of health-care workers.
Over development
the and
past
several of
years,
the
application
molecular
diagnostic techniques has initiated a revolution in the diagnosis and monitoring of infectious diseases.
bacteriophage
chromatographic profiles
biotyping
susceptibility testing
Nucleic
acid
techniques,
such
as
plasmid
clinical laboratories.
PCR-based systems to detect the etiologic agents of disease directly from clinical samples, without the need for culture, have been useful in rapid detection of unculturable or fastidious microorganisms.
Additionally, amplified
sequence DNA
analysis allows
of for
microbial
Other important advances include the determination of viral load and the direct detection of of genes and or gene mutations software widely
In all, the detection of infectious agents at the nucleic acid level represents
At this time, no typing system completely fullls all the criteria described below, and the advantages and disadvantages of a technique should be understood before use Typeability
:
Reproducibility
1.
Typeability: an unambiguous result for each isolate analyzed. Nontypeable isolates have a null or
uninterpretable result.
2. Reproducibility: the same result when the same strain is is tested by repeatedly. technical Reproducibility inuenced
unrelated strains
but varies as species vary in genetic stability under the inuence of selective pressure.
4. Ease of interpretation: multiple users of the typing system obtain the same results and reach the same conclusions.
R C
5. Ease of performance:
the rapidity
convenience,
cost of equipment, and personnel needed to perform a technique .
PHENOTYPE
The difficulty in assigning an organism to a biologically meaningful category should be well considered before the use of any molecular identification tool. Researchers should be aware of the evolutionary history and taxonomic
Ideally, they should understand the order of branching and ages of divergence (phylogeny) of the organisms. Terms such as
However, with the enormous inuence of molecular biology in the 1970s it became increasingly clear that phenotypic
Thus, molecular methods began to nd application in clinically relevant areas including epidemiological analysis, giving rise to the term
molecular epidemiology.
process
the rst iteration of which was analysis of isolate plasmid content by
agarose-gel electrophoresis
However, the bacterial genome is the most fundamental molecule of identity in the cell and thus represents
the
ultimate
metric
for
assessing
isolate
interrelatedness.
It should be realized that a continuous genetic variability does exist among individuals under study has to be properly assessed before
So the choice of a particular technique is often a compromise that depends on a number of factors, including:
the resources of the laboratory. financial constraints. available expertise.
time limitations.
And more importantly, the research question pursued.
RFLP
Ribotyping PCR-RFLP RAPD High Good High High Moderate High High Good Poor Good High High High High Good-High
AFLP
Repetitive elements Sequencing
High
Good
High
Good
Good
High
Moderate
High
High
High
High
High
High
Moderate
Good-High
Schematic representation of most important procedural steps involved in the various DNA-based methods
Specimen Collection
Although viability is not as critical for molecular testing, the quality of nucleic acids may be compromised if the specimen is
Due to the sensitivity of molecular testing, it is also important to avoid contamination that could yield false-positive results
The quantity of target organisms as well as the clinical implications should be taken into account when interpreting the signicance of
First, depending on the microorganism, more rigorous lysis procedures may be required.
Mycobacteria and fungi in particular have thick cell walls that are more difficult to lyse than those of other bacteria and parasites.
Gram-positive bacteria having a thicker cell wall than gram-negative bacteria may require more rigorous cell lysis conditions.
Mycoplasma, on the other hand, lacks a cell wall, and so care must be taken with the sample to avoid spontaneous lysis of the cells and loss of nucleic acids.
Second, the concentration of organisms within the clinical sample must be considered. Samples should be centrifuged to concentrate
the uid.
Third, inhibitors of enzymes used in molecular analysis may be present in clinical specimens; removal or inactivation of inhibitors
Finally,
if
RNA
is
to
be
analyzed,
inactivation or removal of RNases in the sample and in all reagents and materials that come into
Thus,
ensuring
that
the
integrity
of
specimens is maintained, i.e., that specimens are not contaminated by other specimens or
On the other hand, it is equally important to ensure that the lack of a product in an amplication procedure is due to the absence
Finally, DNA can provide more information than proteins due to the degeneracy of the genetic code and the presence of large non-
coding stretches
Future Perspective
High-speed methodological innovation in the
It seems obvious that sequencing will remain the preferred technology for assessing phylogenetic relatedness and will define the
This may generate a novel series of realtime typing procedures of specific relevance to the clinical microbiology laboratory. Such
defined
by
the technology
used but
by
the
timeliness of its data. From that perspective, the expectation is that we will continue to see many changes in the way in which epidemiological data are collected. It remains to be seen; however, which method will answer clinical questions with the highest degree of reliability.