scholarly journals Description of a polyvalent conjugate and a new serogroup of Bacteroides melaninogenicus by fluorescent antibody staining

1976 ◽  
Vol 3 (5) ◽  
pp. 506-512
Author(s):  
D W Lambe ◽  
R C Jerris
Keyword(s):  
Fluorescent Antibody ◽  
Fluorescein Isothiocyanate ◽  
Rapid Identification ◽  
Antibody Staining ◽  
Biochemical Testing ◽  
Fluorescent Antibody Staining

A polyvalent conjugate (fluorescein isothiocyanate-labeled antibody reagent) containing serogroups A, B, and C conjugates was prepared. This polyvalent conjugate gave a positive fluorescent antibody (FA) stain with 49 stains of Bacteroides melaninogenicus representing serogroups A, B, and C. When additional strains (92 strains) of the three subspecies of B. melaninogenicus were examined by the FA stain, with A, B, and C, and polyvalent conjugates, nine strains of B. melaninogenicus subsp. intermedius failed to give a positive stain with any conjugate. Therefore, an FA conjugate was prepared with the antiserum to one of these strains (532-70A); all nine strains stained positively with this conjugate. These nine strains were biochemically characteristic of B. melaninogenicus subsp. intermedius; thus, these strains were designated as a new serogroup, serogroup C-1. A new polyvalent conjugate containing serogroups A, B, C, and C-1 was prepared. This polyvalent conjugate stained positively with 23 representative strains from serogroups A, B, C, and C-1. The new conjugates failed to stain positively with other anaerobes and aerobes tested. The four individual conjugates, as well as the polyvalent conjugate, may be used for a more rapid identification of B. melaninogenicus than is possible by biochemical testing.

Characterization of Bacteroides gingivalis by direct fluorescent antibody staining and cellular fatty acid profiles

1982 ◽  
Vol 28 (4) ◽  
pp. 367-374 ◽  
Author(s):  
Dwight W. Lambe Jr. ◽  
Kaethe P. Ferguson ◽  
William R. Mayberry
Keyword(s):  
Fatty Acid ◽  
Fluorescent Antibody ◽  
Fluorescein Isothiocyanate ◽  
Cellular Fatty Acid ◽  
Fatty Acid Profiles ◽  
Bacteroides Species ◽  
Antibody Staining ◽  
Direct Fluorescent Antibody ◽  
Bacteroides Gingivalis ◽  
Fluorescent Antibody Staining

Bacteroides gingivalis is a newly proposed species which includes strains isolated from the mouth. Thirteen strains of B. gingivalis isolated from three geographic locations in the United States and France were examined with direct fluorescent antibody staining and analysis of total cellular fatty acids and compared with 16 strains of B. asaccharolyticus of nonoral origin by the same methods. Bacteroides gingivalis strains reacted with the B. gingivalis conjugate (fluorescein isothiocyanate labeled antibody reagent) only, while the B. asaccharolyticus strains reacted with the B. asaccharolyticus conjugate only. The B. gingivalis strains showed negative fluorescence with fluorescein isothiocyanate conjugates for other black-pigmented Bacteroides species. The specificity of the B. gingivalis conjugate was demonstrated by its failure to stain 88 strains of aerobic and anaerobic bacteria other than B. gingivalis. The fatty acid profiles of B. gingivalis and B. asaccharolyticus were readily distinguishable. The B. gingivalis profile was also distinguishable from those of other pigmenting Bacteroides species on the basis of concentration ratios among the characteristic components. These results support the species separation of B. gingivalis and B. asaccharolyticus. Further, they indicate the usefulness of cellular fatty acid profiles as an adjunct to the use of specific fluorescent antibody conjugates for identification of Bacteroides species.

Profilin is predominantly associated with monomeric actin in Acanthamoeba

1999 ◽  
Vol 112 (21) ◽  
pp. 3779-3790 ◽  
Author(s):  
D.A. Kaiser ◽  
V.K. Vinson ◽  
D.B. Murphy ◽  
T.D. Pollard
Keyword(s):  
Monoclonal Antibodies ◽  
Plasma Membrane ◽  
Fluorescent Antibody ◽  
Gel Filtration ◽  
Live Cells ◽  
A431 Cells ◽  
Antibody Staining ◽  
Large Particles ◽  
Biochemical Fractionation ◽  
Fluorescent Antibody Staining

We used biochemical fractionation, immunoassays and microscopy of live and fixed Acanthamoeba to determine how much profilin is bound to its known ligands: actin, membrane PIP(2), Arp2/3 complex and polyproline sequences. Virtually all profilin is soluble after gentle homogenization of cells. During gel filtration of extracts on Sephadex G75, approximately 60% of profilin chromatographs with monomeric actin, 40% is free and none voids with Arp2/3 complex or other large particles. Selective monoclonal antibodies confirm that most of the profilin is bound to actin: 65% in extract immunoadsorption assays and 74–89% by fluorescent antibody staining. Other than monomeric actin, no major profilin ligands are detected in crude extracts. Profilin-II labeled with rhodamine on cysteine at position 58 retains its affinity for actin, PIP(2) and poly-L-proline. When syringe-loaded into live cells, it distributes throughout the cytoplasm, is excluded from membrane-bounded organelles, and concentrates in lamellapodia and sites of endocytosis but not directly on the plasma membrane. Some profilin fluorescence appears punctate, but since no particulate profilin is detected biochemically, these spots may be soluble profilin between organelles that exclude profilin. The distribution of profilin in fixed human A431 cells is similar to that in amoebas. Our results show that the major pool of polymerizable actin monomers is complexed with profilin and spread throughout the cytoplasm.

scholarly journals Detection ofCryptosporidium molnariOocysts from Fish by Fluorescent-Antibody Staining Assays forCryptosporidiumspp. Affecting Humans

2011 ◽  
Vol 77 (5) ◽  
pp. 1878-1880 ◽  
Author(s):  
Rona Barugahare ◽  
Michelle M. Dennis ◽  
Joy A. Becker ◽  
Jan Šlapeta
Keyword(s):  
Ribosomal Dna ◽  
Fluorescent Antibody ◽  
Small Subunit ◽  
Rapid Diagnosis ◽  
Antibody Staining ◽  
Direct Fluorescent Antibody ◽  
Murray Cod ◽  
Diagnosis Of Infection ◽  
Formalin Fixed ◽  
Fluorescent Antibody Staining

ABSTRACTThree direct fluorescent-antibody staining assay kits for the detection of zoonoticCryptosporidiumspecies were used to detectCryptosporidium molnarifrom Murray cod, and the cryptosporidia were characterized by using small-subunit (SSU) ribosomal DNA (rDNA). To facilitate rapid diagnosis of infection, this study demonstrated that all three kits detected freshC. molnariand two kits detected formalin-fixed oocysts.

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