Presentation on leishmaniasis

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Upma Gandhi

Gesundheit & Medizin

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Influenza A: The recently raised awareness of the threat of a new influenza pandemic has stimulated interest in the detection of influenza A viruses in human as well as animal secretions. Virus isolation alone is unsatisfactory for this purpose because of its inherent limited sensitivity and the lack of host cells that are universally permissive to all influenza A viruses. Previously described PCR methods are more sensitive but are targeted predominantly at virus strains currently circulating in humans, since the sequences of the primer sets display considerable numbers of mismatches to the sequences of animal influenza A viruses. Therefore, a new set of primers, based on highly conserved regions of the matrix gene, was designed for single-tube reverse transcription-PCR for the detection of influenza A viruses from multiple species. This PCR proved to be fully reactive with a panel of 25 genetically diverse virus isolates that were obtained from birds, humans, pigs, horses, and seals and that included all known subtypes of influenza A virus. It was not reactive with the 11 other RNA viruses tested. Comparative tests with throat swab samples from humans and fecal and cloacal swab samples from birds confirmed that the new PCR is faster and up to 100-fold more sensitive than classical virus isolation procedures. Various Methods of Detection: Specimens. Cloacal swab specimens were collected from ducks (widgeon [Mareca penelope], gadwall [Mareca strepera], and mallard [Anas plathyrhynchos]) at a marshaling lake in Lekkerkerk, The Netherlands, and droppings as well as cloacal swab specimens were collected from geese (greylag goose [Anser anser], white- fronted goose [Anser albifrons albifrons], barnacle goose [Branta leucopsis], and brent goose [Branta bernicla]) in Groningen and Eemdijk, The Netherlands, between 1997 and 1999. Cloacal swab specimens and droppings were collected from shorebirds at Öland, Sweden, in the spring of 1999. Cotton swabs were used for sampling and were subsequently stored in transport medium. Throat swab specimens collected from humans were also stored in transport medium. The samples were stored at 4°C for a few days, at 20°C for less than a week, or at 70°C for extended periods of time. Transport medium consisted of Hanks balanced salt solution supplemented with 10% glycerol, 200 U of penicillin per ml, 200 µg of streptomycin per ml, 100 U of polymyxin B sulfate per ml, 250 µg of gentamicin per ml, and 50 U of nystatin per ml (all from ICN, Zoetermeer, The Netherlands). RNA isolation. RNA was isolated with a high pure RNA isolation kit (Roche Molecular Biochemicals) according to the instructions from the manufacturer, with minor modifications. A 0.2-ml sample was homogenized by vortexing and was subsequently lysed with 0.4 ml of lysis-binding buffer to which poly(A) (Roche Molecular Biochemicals) was added as a carrier to 1 µg/ml. After binding to the column, DNase I digestion, and washing, the RNA was eluted in 50 µl o.

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