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

[Isabel Mauricio]

Dear Katherine

If the trypanosomatids circulate in the blood of infected wildlife, then it should be possible to detect and obtain enough DNA for direct sequencing using sensitive PCR methods, such as nested-PCR, and multicopy targets, such as kDNA and ribosomal ITS/subunits.
Otherwise, it would be best to obtain samples from the organs where the most parasites are expected, such as lymph nodes, bone marrow, skin, etc.

Best regards
Isabel

De: Sayler,Katherine A [mailto:saylerk at ufl.edu]
Enviada: 5 de março de 2015 23:30
Para: leish-l at lineu.icb.usp.br
Assunto: [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


--
This email was sent by icb.usp.br   ­­
-------------- next part --------------
An HTML attachment was scrubbed...
URL: <http://lineu.icb.usp.br/pipermail/leish-l/attachments/20150306/333cb9cc/attachment-0001.htm>