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

Fri Mar 6 05:08:45 BRT 2015

Dear Katherine,
You are completely right that it is difficult to get enough DNA for sequencing from blood, but yes PCR amplification can help get sufficient template  for sequencing (if it is only for a specific gene target). I would suggest to use buffy coat rather than whole blood for PCR. Your results will be dramatically improved.
All the best Prof. Sarman Singh, MBBS, MD, FAMS, FIMSA, FRSTMH, FRSC, FATP, FSIIP
Head, Division of Clinical Microbiology & Molecular Medicine,
All India Institute of Medical Sciences, New Delhi (India)
Editor: Journal of Laboratory PhysiciansPhone: +91-11-26594977, 26588484
Fax: +91-11-26588663, 26588641
https://www.researchgate.net/profile/Prof_Sarman_SinghPersonal Website: www.drsarmansingh.com-BMJ Award 2014, Excellence in Innovative Healthcare Technology
-MedAchiever's Merit Award 2014, Indian Medical Association-Chikitsa Ratan Award 2014, Delhi Medical Association-Dr. BK Aikat Oration 2008, Indian Council of Medical Research-Dr. BC Roy Award 2008, Medical Council of India-Vigyan Ratan Samman, 2007, UP Council of S &T-Dr. BP Pandey Oration Award 2006, Indian Society of Parasitology-Biotech Process, Product  & Commercialization Award, 2006, Department of Biotechnology-Merit Award 1998, International Immnunocompromised Host Society USA-Young Scientist Award 1996, International Immnunocompromised Host Society USA 

     On Friday, March 6, 2015 6:51 AM, "Sayler,Katherine A" <saylerk at ufl.edu> wrote:

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 theLeishmania 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 ConservationMolecular Ecology Laboratory | 2322 Mowry Rd. Gainesville, FL 32611Phone: 352-871-3259 

--
 This email was sent by icb.usp.br     
_______________________________________________
Leish-l mailing list
Leish-l at lineu.icb.usp.br
http://lineu.icb.usp.br/cgi-bin/mailman/listinfo/leish-l

--
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/20150306/78dc3b58/attachment.htm>