PCR-ESI-MS

http://hawaii.gov/health/laboratories/sld-forms/np-2009_AMP_Poster_Leptospirosis_19F.pdf

use broad range PCR primer pairs e.g. for 16S rRNA conserved region
do PCR->generate a mass spectrum->base composition as a signature to determine microbes.

Anti-dsDNA Antibody by immunofluorescence

Crithidia luciliae
•a giant mitochondion containing dsDNA (kinetoplast)
•dose not contain any other antigens present in the cell nuclei

•Indirect immunofluorescence
•Crithidia lucialiae as substrate
•Positive for anti-dsDNA Ab

Biotinidase assay

• Biotinidase cleaves the amide bond of biotinyl-paminobenzoic
acid (B-pABA, an artificial substrate) to
give free biotin and p-aminobenzoic acid (pABA)

• The pABA is diazotized and reacted with a naphthol
reagent to give a purple product which can be
quantitated spectrophotometrically at 546nm

• Biotinidase activity is expressed as nmol of pABA
liberated per minute per ml serum

• Biotinidase deficient cases <0.7 nmol pABA/min/ml