[Leish-l] Confirm hits to Leishmania spp. in whole blood of mammals in the US?

[Bogdan, Christian]

Dear Dr. Sayler,

in proven reservoir hosts (e.g., small mammals) it is no problem to detect Leishmania DNA by PCR in the circulating blood if these animals are infected. However, one has to be very cautious with the selection of primers. For example, PCR methods based on primers targeting the small subunit rRNA gene or the 18S rRNA gene can yield positive results, but sequencing of the PCR products reveal the presence of apathogenic Trypanosomes rather than Leishmania.

With regards,

Christian Bogdan


Christian Bogdan, M.D.
Professor of Medical Microbiology and Infectious Disease Immunology, Head of the Institute
Microbiology Institute - Clinical Microbiology, Immunology and Hygiene
Universitätsklinikum Erlangen
Friedrich-Alexander Universität Erlangen-Nürnberg
Wasserturmstraße 3/5
D-91054 Erlangen, Germany
Tel. (+49)-9131-852-2551 (office), -2281 (secretary)
Fax (+49)-9131-852-2573 or -9131-85-1001
E-mail: christian.bogdan at uk-erlangen.de<mailto:christian.bogdan at uk-erlangen.de>
http://www.mikrobiologie.uk-erlangen.de/


Von: leish-l-bounces at lineu.icb.usp.br [mailto:leish-l-bounces at lineu.icb.usp.br] Im Auftrag von Sayler,Katherine A
Gesendet: Freitag, 6. März 2015 00:30
An: leish-l at lineu.icb.usp.br
Betreff: [Leish-l] Confirm hits to Leishmania spp. in whole blood of mammals in the US?


Dear readers,



Thank you sincerely for your time. I am a tick-borne disease biologist by trade, so this is outside of my realm.



I am trying to confirm some preliminary data that suggests the Leishmania spp. are circulating in limited geographic locales in southern Florida. Is it at all reasonable to expect to detect the DNA of the trypanosomatids in the blood of infected wildlife? My suspicion is that this is completely improbable, as it appears that in order to detect DNA in enough quantity or quality for sequencing, you need cultured organisms. From sandflies it appears that it is possible to PCR amplify and sequence directly from the vector, but never from the reservoir (or potential reservoir) host. Is this true? Any advice or insight would be greatly appreciated and I am tremendously naïve and this is clearly not my area of expertise.



Thank you again for reading my message,

Katherine

Katherine Sayler, M.Ed., Ph.D.
Postdoctoral Associate | Department of Wildlife Ecology and Conservation
Molecular Ecology Laboratory | 2322 Mowry Rd. Gainesville, FL 32611
Phone: 352-871-3259


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