antibodies against whole bacteria - dectection problems in plasma

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osiris-gdw
osiris-gdw's picture
antibodies against whole bacteria - dectection problems in plasma

Hello,

I have to detect life bacteria in human plasma with antibodies!

Does anyone know a real good, and high concentrated antibody against Staphylococcus aureus or E. coli
and do you know of detection problems in human plasma/serum?

Thanks in advance!

Regards,

Gerhard

Tony Rook
Tony Rook's picture
Hello Gerhard:

Hello Gerhard:

Here are some references that should help you out...

Miwa K, Fukuyama M, Sakai R, Shimizu S, Ida N, Endo M, Igarashi H. Sensitive enzyme-linked immunosorbent assays for the detection of bacterial superantigens and antibodies against them in human plasma. Microbiol Immunol. 2000;44(6):519-23.

Abstract:
Enzyme-linked immunosorbent assays for the quantitation of bacterial superantigens, staphylococcal enterotoxins A, B and C, toxic-shock syndrome toxin-1 and streptococcal pyrogenic exotoxin A, were developed. The assays had sensitivity to quantitate these toxins to 1.4, 5.9, 16.3, 2.5 and 4.3 pg/ml, respectively, in a buffer including 50% human plasma. It takes only 150 min to complete the assays after plate preparation. Specificity of the assays agreed with those of reverse latex agglutination assay. We also developed enzyme-linked immunosorbent assays to detect antibodies against these five superantigens. The assays are expected to be significant tools for the study of superantigens in several diseases.

K. Matsubara, T. Fukaya, K. Miwa, N. Shibayama, H. Nigami, H. Harigaya, H. Nozaki, T. Hirata, K. Baba, T. Suzuki, A. Ishiguro (2006) Development of serum IgM antibodies against superantigens of Staphylococcus aureus and Streptococcus pyogenes in Kawasaki disease. Clinical & Experimental Immunology 143 (3), 427434.
doi:10.1111/j.1365-2249.2006.03015.x
pdf link

Abstract:
To serologically determine the association of microbial superantigens and the pathogenesis of Kawasaki disease (KD), we conducted a case-control study. Serum IgG and IgM antibodies against staphylococcal enterotoxin A (SEA), SEB, SEC, toxic shock syndrome toxin-1 (TSST-1), and streptococcal pyrogenic exotoxin A (SPEA) were measured by an enzyme-linked immunosorbent assay in 293 serum samples from 65 KD patients on clinical days 128 and 120 control samples. The administration of immunoglobulin products, which contain high concentrations of IgG antibodies against all the superantigens, directly elevated antitoxin IgG antibodies in KD patients. In contrast, antitoxin IgM antibodies were not detected in immunoglobulin products. Actually, we found a significant elevation of IgM antibodies against SEA in KD patients in the first (median titre: 0020, P < 001 versus control), second (0024, P < 0001), third (0030, P < 0001) and fourth (0038, P < 0001) weeks, compared to the controls (0015). Significant differences of IgM antibodies were also true for SEB, TSST-1, and SPEA throughout the first to fourth weeks, and for SEC throughout the second to fourth weeks. The prevalence of KD patients having high IgM titres (> mean + 2SD of control values) to the 5 superantigens was increased with the clinical weeks, and reached 2943% of KD subjects at the fourth week. This is the first study that describes kinetics of IgM antibodies against superantigens and clarifies the serological significance throughout the clinical course of KD. Our results suggest that multiple superantigens involve in the pathogenesis of KD.

Shoshana Bascomb and Mammad Manafi Use of Enzyme Tests in Characterization and Identification of Aerobic and Facultatively Anaerobic Gram-Positive Cocci. Clinical Microbiology Reviews, April 1998, p. 318-340, Vol. 11, No. 2
pdf link

Abstract:
The contribution of enzyme tests to the accurate and rapid routine identification of gram-positive cocci is introduced. The current taxonomy of the genera of aerobic and facultatively anaerobic cocci based on genotypic and phenotypic characterization is reviewed. The clinical and economic importance of members of these taxa is briefly summarized. Tables summarizing test schemes and kits available for the identification of staphylococci, enterococci, and streptococci on the basis of general requirements, number of tests, number of taxa, test classes, and completion times are discussed. Enzyme tests included in each scheme are compared on the basis of their synthetic moiety. The current understanding of the activity of enzymes important for classification and identification of the major groups, methods of testing, and relevance to the ease and speed of identification are reviewed. Publications describing the use of different identification kits are listed, and overall identification successes and problems are discussed. The relationships between the results of conventional biochemical and rapid enzyme tests are described and considered. The use of synthetic substrates for the detection of glycosidases and peptidases is reviewed, and the advantages of fluorogenic synthetic moieties are discussed. The relevance of enzyme tests to accurate and meaningful rapid routine identification is discussed.

J M Fournier, A Bouvet, D Mathieu, F Nato, A Boutonnier, R Gerbal, P Brunengo, C Saulnier, N Sagot and B Slizewicz. New latex reagent using monoclonal antibodies to capsular polysaccharide for reliable identification of both oxacillin-susceptible and oxacillin-resistant Staphylococcus aureus. J Clin Microbiol. 1993 May; 31(5): 1342-1344
pdf link

Abstract:
A new latex agglutination test (Pastorex Staph-Plus, Sanofi Diagnostics Pasteur), consisting of a mixture of latex particles coated with fibrinogen and immunoglobulin G for the detection of clumping factor and protein A and latex particles sensitized with monoclonal antibodies directed to Staphylococcus aureus serotype 5 and 8 capsular polysaccharides, was compared with three commercially available rapid agglutination methods for the identification of 220 isolates of S. aureus (61 oxacillin resistant) and 128 isolates of coagulase-negative staphylococci. The sensitivity for identification of S. aureus was high with the Pastorex Staph-Plus test (98.6%) compared with those of the other tests, which ranged from 91.8 to 84.5%. Test sensitivities for the identification of oxacillin-resistant S. aureus were as follows: Pastorex Staph-Plus, 95.1%; Pastorex Staph, 73.8%; Staphyslide, 72.1%; and StaphAurex, 49.2%.

Jason King
Jason King's picture
osiris-gdw wrote:Hello,

osiris-gdw wrote:

Hello,

I have to detect life bacteria in human plasma with antibodies!

Does anyone know a real good, and high concentrated antibody against Staphylococcus aureus or E. coli
and do you know of detection problems in human plasma/serum?

Thanks in advance!

Regards,

Gerhard

I guess you will be able to find a specific antibody but in case you don't and want to try something slightly different:

A colleague of mine is using fluorescently tagged phages to detect bacteria. He is using staph. aureus and has a phage that is specific for it.
He is interested in detecting the live staph and looking for colocalization of these and GFP expressing macrophages in-vivo as the macs take phagocytose the bugs. The drawback is that the phages kill the staph as they are lytic. I thought it was a nice alternative to bug-specific fluorescent antibodies.