proposed a 2-tiered EIA approach consisting of 2 FDA-cleared EIAs: whole-cell sonicate
EIA followed by reflex C6 EIA.
miyamotoi (30), no data are available that examined the potential cross-reactivity of whole-cell sonicate
or C6 B.
Since the cross-reacting material is PorA, it is present in sufficient quantity in bacterial sonicates
used in the direct ELISA for all strains to be positive.
Extraction Extraction First water Sample type solvent (a) method addition Air filter 5 mL x 2 Sonicate
100 mL Air PUF 30 mL x 4 Squeeze 80 mL Impactor plate 5 mL x 2 Sonicate
100 mL Impactor filter 10 mL x 2 Sonicate
100 mL Surface wipe 20 mL x 2 Sonicate
360 mL PUF roller sleeve 150 mL x 4 Squeeze 150 mL Floor dust (bulk) 25 mL Sonicate
100 mL Hexane Second water Sample type partition addition Air filter 20 mL x 2 100 mL Air PUF 20 mL x 2 70 mL Impactor plate 20 mL x 2 100 mL Impactor filter 20 mL x 2 100 mL Surface wipe 20 mL x 2 0 mL PUF roller sleeve 25 mL x 2 0 mL Floor dust (bulk) 20 mL x 2 80 mL Variation in standard procedure, scaled to size of sample matrix.
The SONOPULS model homogenizers sonicate
volumes from 50 ml up to 11 on a batch basis.
burgdorferi by using a whole-cell sonicate
ELISA, C6 ELISA, or Western blot assay as described (16,20-22).
Immunoaffinity chromatographic isolation of a high molecular weight seroreactive protein from Mycobacterium leprae cell sonicate
Th2-type immune response observed in healthy individuals to sonicate
antigen prepared from the most prevalent Mycobacterium tuberculosis strain with single copy of IS6110.
9%) to dissolve and sonicate
for about 5 min, dilute to volume with methanol and mix.
For preparation of triton-soluble extracts, the sonicate
from each sample was diluted with 1x PBS containing protease inhibitor and Triton X-100 (Loba Chemie, Mumbai, India) (1% final concentration) up to 200 [micro]l at 5 [micro]g/[micro]l protein concentration and mixed thoroughly for complete solubilization (12).
Samples obtained from these patients were tested by a commercial enzyme-linked immunosorbent assay (ELISA) (PanBio, East Brisbane, Queensland, Australia) for the presence of immunoglobulin (Ig) A against a Bordetella pertussis sonicate
and by an in-house IgA-detection ELISA directed against pertussis toxin as previously described (9).
GSTA1-1 was purified from the bacterial sonicate
by gel filtration on Sephadex G50 (Pharmacia) followed by affinity chromatography on S-hexylglutathione (35) coupled to epoxy-activated Sepharose 6B (Pharmacia).