A Manual of Clinical DiagnosisTodd, James Campbell
Science
A Manual of Clinical Diagnosis
Todd, James Campbell
Diagnosis, Laboratory
1. Agglutination.--In the blood-serum of persons suffering from
certain infectious diseases there exist soluble bodies, called
agglutinins, which have the property of {197} rendering non-motile and
clumping the specific micro-organism of the disease, and have little
or no influence upon other bacteria. This "agglutination" takes place
even when the blood is greatly diluted. Undiluted normal blood can
agglutinate most bacteria, but loses this power when diluted to any
considerable degree. These facts are taken advantage of in the
diagnosis of several diseases.
When applied to the diagnosis of typhoid fever, the phenomenon is
known as the _Widal reaction_. As yet, it is the only agglutination
reaction which has any practical value for the practitioner.
Either blood-serum or the whole blood may be used. Serum is the
better. To obtain it, it is convenient to use little vials, such as
can be made by breaking off the lower half-inch of the tubes which
have contained peptonizing powder. They must, of course, be well
cleaned. One of these is filled to a depth of about one-fourth inch
from a puncture in the finger, and is set aside for a few hours. When
the clot has separated, it is picked out with a needle, leaving the
serum. One drop of the serum is then added to nine drops of normal
salt solution, making a dilution of 1:10. Distilled water may be used
for dilution, but is more liable to cause error. The dilution can be
more accurately made in the leukocyte pipet of the Thoma-Zeiss
instrument. When the whole blood is used, it can be secured in the
pipet and at once diluted with the salt solution. When it must be
transported a considerable distance, dried blood is most convenient. A
large drop is allowed to dry upon a clean slide or unglazed paper. It
will keep for months without losing its agglutinating power. When
ready to make the test, the dried stain is dissolved in ten drops of
normal salt solution, care being {198} taken that the drops are about
the same size as the original drop of blood.
The reaction can be detected either microscopically or
macroscopically:
* * * * *
Microscopic Method.--(1) The blood or serum having been obtained and
diluted 1:10 as just described, mix it with a bouillon culture of the
typhoid bacillus to any desired dilution. One drop of each makes a
blood-dilution of 1:20, etc. The culture should be between eighteen
and twenty-four hours old, and the bacilli must be actively motile. A
stock agar culture should be kept at room temperature, and bouillon
tubes inoculated the day before the examination is to be made. Agar
cultures can be purchased from dealers in biologic products. They must
be renewed monthly.
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