DIAGNOSING

INFECTIOUS
DISEASES
Chapter 13
The doctor, nurse, medical technologist, or other qualified
healthcare professional must select the appropriate
specimen, collect it properly, and then properly transport
it to the CML(Clinical Microbiology Laboratory) for
processing.
Proper Selection, Collection, and
Transport of Clinical Specimens
When collecting clinical specimens for microbiology, these
general precautions should be taken:
• The specimen must be properly selected. That is, it must be the
appropriate type of specimen for diagnosis of the suspected
infectious disease.
• The specimen must be properly and carefully collected.
Whenever possible, specimens must be collected in a manner that
will eliminate or minimize contamination of the specimen with
indigenous microflora
• The material should be collected from a site where the
suspected pathogen is most likely to be found and where
the least contamination is likely to occur.
• Whenever possible, specimens should be obtained
before antimicrobial therapy has begun. If this is not
possible, the laboratory should be informed as to which
antimicrobial agent(s) the patient is receiving.
• The acute stage of the disease—when the patient is
experiencing the symptoms of the disease—is the
appropriate time to collect most specimens. Some viruses,
however, are more easily isolated during the prodromal or
onset stage of disease.
• Specimen collection should be performed with care and tact to
avoid harming the patient, causing discomfort, or causing undue
embarrassment. If the patient is to collect the specimen, such as
sputum or urine, the patient must be given clear and detailed
collection instructions.
• A sufficient quantity of the specimen must be obtained to
provide enough material for all required diagnostic tests. The
amount of specimen to collect should be specified in the Lab P&P
Manual.
• All specimens should be placed or collected into a sterile
container to prevent contamination of the specimen by indigenous
microflora and airborne microbes. Appropriate types of collection
devices and specimen containers should be specified in the Lab
P&P Manual.
• Specimens should be protected from heat and cold and
promptly delivered to the laboratory so that the results of
the analyses will validly represent the number and types
of organisms present at the time of collection. If delivery
to the laboratory is delayed, some delicate pathogens
might die; therefore, certain types of specimens must be
rushed to the laboratory immediately after collection.
Some types of specimens must be placed on ice
during delivery to the laboratory, whereas other
specimens should never be refrigerated or placed on
ice because of the fragile and sensitive nature of the
pathogens. Obligate anaerobes die when exposed to
air and therefore must be protected from oxygen
during transport to the CML. Any indigenous
microflora in the specimen may overgrow, inhibit, or
kill pathogens. Specimen transport instructions should
be contained in the Lab P&P Manual.
• Specimens must be handled with great care to avoid
contamination of the patients, couriers, and healthcare
professionals. Specimens must be placed in a sealed plastic
bag for immediate and careful transport to the laboratory.
Whenever possible, sterile, disposable specimen containers
should be used. • The specimen container must be properly
labeled and accompanied by an appropriate laboratory test
requisition containing adequate instructions. Labels should
contain the patient’s name, unique hospital identification
number, and hospital room number; requesting clinician’s
name; culture site; and date and time of collection.
Laboratory test requisitions should contain the patient’s
name, age, sex, and unique hospital identification
number; name of the requesting clinician; specific
information about the type of specimen and the site
from which it was collected; date and time of
collection; initials of the person who collected the
specimen; and information about any antimicrobial
agent(s) that the patient is receiving. The laboratory
should always be given sufficient clinical information
to aid in performing appropriate analyses.
For example, the laboratory test requisition that
accompanies a wound specimen should not merely state
“wound”; rather, it should state the specific type of
wound (e.g., burn wound, dog bite wound, surgical site
infection, etc.), the anatomical site, and whether it is on
the left side or right side, if applicable. • Ideally,
specimens should be collected and delivered to the
laboratory as early in the day as possible to give CML
professionals sufficient time to process the material,
especially when the hospital or clinic does not have 24-
hour laboratory service.
The various phenotypic characteristics (clues) useful in identifying bacteria
include the following:
• Gram reaction (i.e., Grampositive or Gram-negative)
• Cell shape (e.g., cocci, bacilli, curved, spiral-shaped, filamentous,
branching)
• Morphologic arrangement of cells (e.g., pairs, tetrads, chains, clusters) •
Growth or no growth on various types of plated media
• Colony morphology (e.g., color, general shape, elevation, margin)
• Presence or absence of a capsule
• Motility
• Number and location of flagella
• Ability to sporulate
• Location of spores (terminal or subterminal)
• Presence or absence of various enzymes (e.g., catalase,
coagulase, oxidase, urease)
• Ability to catabolize various carbohydrates and amino acids
(miniaturized biochemical test systems— “minisystems”—are
often used for this purpose; see Fig. 13-12)
• Ability to reduce nitrate
• Ability to produce indole from tryptophan
• Atmospheric requirements
• Type of hemolysis produced (Fig. 13-13)