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

immobilized metal-affinity chromatography (IMAC)

IMAC is based on the interactions between a transition metal ion (Co2+, Ni2+, Cu2+, Zn2+) immobilized on a matrix and specific amino acid side chains. Histidine is the amino acid that exhibits the strongest interaction with immobilized metal ion matrices, as electron donor groups on the histidine imidazole ring readily form coordination bonds with the immobilized transition metal.

e.g. Polyhistidine-tag column

Jenner's Stain

Jenner's Stain (methylene blue eosinate) is used in microscopy for staining blood smears.

Wright's stain

Wright's stain is a histologic stain that facilitates the differentiation of blood cell types. It is used primarily to stain peripheral blood smears and bone marrow aspirates which are examined under a light microscope. In cytogenetics it is used to stain chromosomes to facilitate diagnosis of syndromes and diseases.

Warthin–Starry stain

The Warthin–Starry stain (WS) is a silver nitrate-based staining method (a silver stain) used in histology. It was first introduced in 1920 by American pathologists Aldred Scott Warthin (1866-1931) and Allen Chronister Starry (1890-1973), for the detection of spirochetes.[1][2] It has been considered the best stain for detection of spirochetes,[3] and is also used to stain Helicobacter pylori, Lawsonia intracellularis,[4] Microsporidia,[5][6] and particulates.[7]
WS stains organisms dark brown to black, and the background light golden brown/golden yellow.