Medical Microbiology

cubatedinvitrowitherythrocytesorantigenicparticles.Inallcasesagglutina-
tionisdetectedusinganti-Igantibodies.Antigenscanalsobeadsorbedtolatex.

ComplementFixationTest(CFT)......................

CFTwasformerlyusedtomeasurecomplementconsumptionbypreformed
antigen-antibodycomplexes.Theunusedcomplementisthendetectedbyad-
ditionofaknownamountofantibody-loadederythrocytes.Shouldallofthe
erythrocytesbelysed,thisindicatesthatnocomplementhadbeenconsumed
andtheCFTisnegative.Thismethodisnolongerusedveryfrequently,with
thenewerimmunosorbenttestsbeingpreferred(RIA,ELISA,RAST,seebelow).

DirectandIndirectImmunofluorescence.............

Directimmunofluorescence.Immunofluorescencecanbeusedforin-vivo
detectionofantibodies,complement,viruses,fungi,bacteria,orotherim-

ImmunologicalTestMethods 125

WesternBlotting

Fig.2. 24 Antigensamplesseparatedinagelaretransferredtonitrocellulose.

Non-specificbindingoftheantibodiestothefilteristhenpreventedwithserum

albuminorirrelevantproteinsthatdonotcross-reactwithanyoftheantibodies

used.Antibodiesspecificfortheantigensbeingsoughtarethenadded.Onceim-

munecomplexeshaveformed,theunboundantibodiesarethoroughlywashed

awayandtheremainingboundantibodiesarelabeledusinganti-immunoglobulin

antibodies.Theseareinturnrenderedvisiblebytheautoradiographicprocedure.

2

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Kayser, Medical Microbiology © 2005 Thieme

All rights reserved. Usage subject to terms and conditions of license.