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12039759
Detection and enumeration of Cryptosporidium parvum in both treated and untreated waters are important to facilitate prevention of future cryptosporidiosis incidents. Immunomagnetic separation (IMS)-fluorescent antibody (FA) detection and IMS-PCR detection efficiencies were evaluated in two natural waters seeded with nominal seed doses of 5, 10, and 15 oocysts. IMS-FA detected oocysts at concentrations at or below the three nominal oocyst seed doses, illustrating that IMS-FA is sensitive enough to detect low oocyst numbers. However, the species of the oocysts could not be determined with this technique. IMS-PCR, targeting the 18S rRNA gene in this study, yielded positive amplification for 17 of the 18 seeded water samples, and the amplicons were subjected to restriction fragment length polymorphism digestion and DNA sequencing for species identification. Interestingly, the two unseeded, natural water samples were also PCR positive; one amplicon was the same base pair size as the C. parvum amplicon, and the other amplicon was larger. These two amplified products were determined to be derived from DNA of Cryptosporidium muris and a dinoflagellate. These IMS-PCR results illustrate that (i) IMS-PCR is able to detect low oocyst numbers in natural waters, (ii) PCR amplification alone is not confirmatory for detection of target DNA when environmental samples are used, (ii) PCR primers, especially those designed against the rRNA gene region, need to be evaluated for specificity with organisms closely related to the target organism, and (iv) environmental amplicons should be subjected to appropriate species-specific confirmatory techniques.
Immunomagnetic Separation (IMS)-Fluorescent Antibody Detection and IMS-PCR Detection of Seeded Cryptosporidium parvum Oocysts in Natural Waters and Their Limitations
Gregory D. Sturbaum,1,2 Patricia T. Klonicki,2 Marilyn M. Marshall,1 B. Helen Jost,1 Brec L. Clay,2 and Charles R. Sterling1,*
2002 Jun
21581965
In the title compound, C26H25NO6, the anthraquinone ring system forms a dihedral angle of 15.5?(1)° with the benzene ring of the dimethylaniline group. Intramolecular O?H?O hydrogen bonding is observed between the carbonyl and two hydroxyl groups. The molecules are linked into a ribbon-like structure along the [100] direction by O?H?N and C?H?O hydrogen bonds. The crystal used was twinned via a 180° rotation about [100]. The ratio of the two twin components is 0.947?(1):0.053?(1).
6-[3-(2,4-Dimethylanilino)-2-hydroxypropoxy]-1,8-dihydroxy-3-methyl-9,10-dihydroanthracene-9,10-dione
Xing-Po Wanga,* and Wenfang Xub
2009 Feb 1;
23476327
The title compound, [Ru(C7F5O2)2(C10H14)(C21H24N2)]·2CH2Cl2, is formed as an orange crystalline powder by the reaction of RuCl2(p-cymene)(IMes) and AgOCOC6F5 in anhydrous tetrahydrofuran (IMes = 1,3-dimesityl-2,3-dihydro-1H-imidazol-2-ylidene). The asymetric unit consists of two independent [Ru(C6F5COO)2(η6-p-cymene)(IMes)] complexes and four dichloromethane solvent molecules. In each complex molecule, the Ru atom presents a pseudo-octahedral environment with the p-cymene ligand occupying three facial coordination sites, while the remaining coordination positions are occupied by the O atoms of the pentafluorobenzoate ligands and by the imidazolylidene ligand.
(η6-p-Cymene)(1,3-dimesityl-2,3-dihydro-1H-imidazol-2-ylidene)bis(pentafluorobenzoato-κO)ruthenium(II) dichloromethane disolvate
Konstantin Dorst,a Wolfgang Frey,b Dongren Wang,a and Michael Buchmeisera,*
2013 Jan 1;
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