Clostridium Assay

Bouza E, Pelaez T, Alonso R, Catalan P, Munoz P, Creixems MR Second-look cytotoxicity An evaluation of culture plus cyto-toxin assay of Clostridium difficile isolates in the laboratory diagnosis of CDAD. J Hosp Infect 2001 48 233-237. 

The detection of flu viruses via a fluorescent sandwich immunoassay was reported by Bucher.(10) However, the method sensitivity was too low for direct detection of the virus. A novel sandwich immunoassay was described by Ogcr((lff7 for the detection of Botulinum Toxin A. Antibodies specific for Clostridium botulinum were covalently attached to the surface of a tapered fiber. After the capture of the antigen, a sandwich was formed with a rhodamine-labeled anti-toxin IgG, and the evanescent wave was measured. The assay was highly specific with detection limits near 5 ppb. 

Fach, P., Perelle, S., Dilasser, F., Grout, J., Dargaignaratz, C., Botella, L., Gourreau, J.-M., Carlin, F., Popoff, M.R. and Broussolle, V., Detection by PCR-enzyme-linked immunosorbent assay of Clostridium botulinum in fish and environmental samples from a coastal area in Northern France, Appl. Em. Microbiol., 68, 5870-5876, 2002. 

A 66-year-old man is admitted to the intensive care unit of a hospital for treatment of community-acquired pneumonia. He receives ceftriaxone and azithromycin upon admission, rapidly improves, and is transferred to a semiprivate ward room. On day 7 of his hospitalization, he develops copious diarrhea with eight bowel movements that day but is other wise clinically stable. Clostridium difficile-associated colitis is suspected and a toxin assay is sent to confirm this diagnosis. What is an acceptable treatment for the patient s diarrhea The patient is transferred to a single-bed room the following day. The housekeeping staff asks if the old room should be cleaned with alcohol or bleach. Which product should be chosen Why ... 

Clostridium botulinum neurotoxin, the most effective toxin known to date, with a mice lethal dose of about 50 pg/mL (330 fmol/mL) was the target antigen in IPCR assays developed by Wu et al. [48] and Chao et al. [88]. In these assays, detection limits of 5 fg (33 amol) and 50 fg (330 amol), respectively, were found. 

Chao HY, Wang YC, Tang SS, Liu HW. A highly sensitive immuno-polymerase chain reaction assay for Clostridium botulinum neurotoxin type A. Toxicon 2004 43(1) 27—34. 

Fig. 4. H2 photoevolution from ascorbate via P. laminosum PSI particles under different conditions of immobilization. The complete assay system contained 100 mM Mes-NaOH buffer, pH 7.0 75 mM sodium ascorbate 15 mM DTT 2 mM TMPD 1% (w/v) BSA PSI particles (30 pg Chi) 50 pi (saturating amount) of Clostridium pasteurianum hydrogenase and 12.5 pM Spirulina maxima Fd as electron mediator, (a) Conditions all components free (b) hydrogenase immobilized in Ca alginate according to Gisby and Hall (1980) (c) PSI particles immobilized in Ca alginate (d) hydrogenase and PSI coimmobilized in Ca alginate.

The activities of intracellular enzymes give an indication of the activity of certain metabolic pathways. They are classically assayed by performing their specific reactions in vitro, using the conversion of natural or artificial substrates and often some additional detection reaction. The sample preparation procedure involves cell harvesting and preparation of cell extracts. A well-studied example is the metabolic shift in Clostridium acetobutylicum from acid to solvent production, the so-called solvent shift. Andersch et al. [48] studied the activities of 10 different enzymes involved in this shift, in batch cultivations where the shift is self-induced by the products formed, and under continuous culture conditions with an externally induced shift. 

Clostridium difficile can be cultured from the stool, and toxins A and B can be assessed by different techniques (116). The most accurate method is still a cytotoxin tissue culture assay. This detects the cytopathic effect of cytotoxin B, which can be neutralized by Clostridium sordellii antitoxin, but it takes 24 8 hours to show a result. Alternative tests that produce faster results have been developed. A latex agglutination test lacks sensitivity and specificity, and does not distinguish toxigenic from non-toxigenic strains. An enzyme immunoassay for toxin A may be an acceptable alternative to the cell cytotoxin assay and the results are rapidly available. A dot immunobinding assay has not yet been extensively studied (164). 

Woods GL, Iwwen PC. Comparison of a dot immunobind-ing assay, latex agglutination, and cytotoxin assay for laboratory diagnosis of Clostridium difficile-associated diarrhea. J Clin Microbiol 1990 28(5) 855-7. 

An 85-year-old woman developed a severe illness (severe diarrhea and vomiting, abdominal tenderness, peritoneal irritation, and systemic toxicity) 8 days after receiving pristinamycin 3g/day for 10 days (34). An assay for Clostridium difficile was positive. She was treated with metronidazole and her symptoms resolved after 72 hours. 

The essential oil from another Achillea species, Achillea sintenisii, was assayed for its antimicrobial activities against 12 bacteria and 2 yeasts [26]. The oil was found to be active against some of the test microorganisms studied. The analysis of the oil revealed that the main components, e.g., camphor. Fig. (1) and eucalyptol, possessed appreciable activity against Candida albicans and Clostridium perfringens. The antimicrobial activities of the essential oils of Achillea setacea W. K. and Achillea teretifolia Willd. were individually evaluated against 14... 

Rapid diagnostic assays for detection of toxin agents are available for Botulinum Toxin Clostridium Perfringens Toxin Staphylococcal Enterotoxin B and Staphylococcal Enterotoxins A/C1,2,3/D... 

Sharma, S.K., Ferreira, J.L., Eblen, B.S., and Whiting, R.C. 2006. Detection of type A, B, E, andF Clostridium botulinum neurotoxins in foods by using an amphfied enzyme-hnked immunosorbent assay with digoxigenin-labeled antibodies. Appl. Environ. Micorbiol. 72 1231-1248. 

As well as the enzyme 3a-hydroxysteroid dehydrogenase, a 7a-enzyme has been isolated from E. coli and a 12a enzyme from a strain of Clostridium (Ml). These group-specific enzymes can be used to measure the amounts and ratios of the three main bile acids (cholic, chenodeoxycholic, and deoxy-cholic acids) in bile specimens. Bioluminescent assays suitable for serum bile acid analysis have been described using 7a-hydroxysteroid dehydrogenase (R6) and 12a-hydroxysteroid dehydn enase (S12), as well as for the 3a enzyme (S13, S44). 

Karube et al. (1977c) proposed a biofuel cell containing Clostridium butyricum cells for BOD assay. The anodic oxidation current of hydrogen and formate anaerobically formed from organic compounds was used as the measuring signal. 

Rapid diagnostic assays fielded in support Botulinum Toxin of Operation Desert Stonn/Shieldk Clostridium Perfringens Toxin... 

Recently IHC has been successfully used to identify Streptococcus pneumoniae in formalin-fixed organs with an overall sensitivity of 100% and a specificity of 71% when compared with cultures.Immunohistochemical assays are used to identify Clostridium sp., S. aureus, and S. pyogenes Haemophilus influenzae Chlamydia species i i Legionella pneumophila and L. dumoffiif Listeria monocytogenesf Salmonellaand rickettsial infections other than Rocky Mountain spotted fever such as boutonneuse fever, epidemic typhus, murine typhus,rickettsialpox, 489 African tick bite fever,i and scrub typhus. 

UK-69,753 (SMI) was isolated from cultures of Amycolatopsis orientalis [265]. Its structure was determined using UV, FABMS, elemental analysis, H and NMR, acid hydrolysis to give the disaccharide, and X-ray crystallography of the disaccharide unit [266], In vitro assay showed 90 posesscd antibacterial activity, being particularly effective against Clostridium difficile and Treponema hyodysenteriae (MIC = 0.39 pg/ml and 0.78 pg/ml, respectively [266], In vivo, 90 (3.6 or 7.1 mg/kg/day) provided effective treatment of mice colonized with T. hyodysenteriae [266]. 

Hsieh, H.V., Stewart, B., Hauer, P., Haaland, P., and Campbell, R. (1998) Measurement of Clostridium perfringens (3 toxin production by surface plasmon resonance immuno assay. Vaccine, 16, 997 1003. 

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