<html><body><div style="color:#000; background-color:#fff; font-family:HelveticaNeue, Helvetica Neue, Helvetica, Arial, Lucida Grande, sans-serif;font-size:12pt"><div><span>The one by Reale is the 13A/13B kDNA protocol. Its extremely sensitive. We've tried it. The problem is that after sometime, one start to see that its becoming positive for all samples. Then we gave it up for the benefit of genomic-based methods. Perhaps we should have used UNG-uracil in the PCR setup as an anti-contamination agent.</span></div><div style="color: rgb(0, 0, 0); font-size: 16px; font-family: HelveticaNeue,Helvetica Neue,Helvetica,Arial,Lucida Grande,sans-serif; background-color: transparent; font-style: normal;"><br><span></span></div><div style="color: rgb(0, 0, 0); font-size: 16px; font-family: HelveticaNeue,Helvetica Neue,Helvetica,Arial,Lucida Grande,sans-serif; background-color: transparent; font-style: normal;"><span>Amer</span></div><div style="color: rgb(0, 0,
0); font-size: 16px; font-family: HelveticaNeue,Helvetica Neue,Helvetica,Arial,Lucida Grande,sans-serif; background-color: transparent; font-style: normal;"><span>Palestine<br></span></div><div style="color: rgb(0, 0, 0); font-size: 16px; font-family: HelveticaNeue,Helvetica Neue,Helvetica,Arial,Lucida Grande,sans-serif; background-color: transparent; font-style: normal;"><span><br></span></div><div> </div><div><br></div><div><br></div> <div style="font-family: HelveticaNeue, Helvetica Neue, Helvetica, Arial, Lucida Grande, sans-serif; font-size: 12pt;"> <div style="font-family: HelveticaNeue, Helvetica Neue, Helvetica, Arial, Lucida Grande, sans-serif; font-size: 12pt;"> <div dir="ltr"> <hr size="1"> <font face="Arial" size="2"> <b><span style="font-weight:bold;">From:</span></b> Stefano Reale <stefano.reale@izssicilia.it><br> <b><span style="font-weight: bold;">To:</span></b> Kwang-Poo Chang <kwangpoo.chang@rosalindfranklin.edu>
<br><b><span style="font-weight: bold;">Cc:</span></b> "Lawyer, Phillip (NIH/NIAID) [E]" <PhillipL@niaid.nih.gov>; "leish-l-bounces@lineu.icb.usp.br" <leish-l-bounces@lineu.icb.usp.br>; Rossella Colomba Lelli <rossella.lelli@izssicilia.it>; Leish-L <Leish-L@lineu.icb.usp.br>; "caporalevincenzo@gmail.com" <caporalevincenzo@gmail.com> <br> <b><span style="font-weight: bold;">Sent:</span></b> Thursday, October 31, 2013 8:19 AM<br> <b><span style="font-weight: bold;">Subject:</span></b> Re: [Leish-l] why is Leishmania donovani restricted to humans in Indian subcontinent?<br> </font> </div> <div class="y_msg_container"><br><div id="yiv2080313465"><div><div>Hallo Im one of the author of this PCR protocol. I optimised it by specific primers and probe for infantum. It work very well and I showed the Lod and Loq in variuos biological material taken from leishmaniotic dogs. In my lab in Palermo I have a number if collected strains
stored in liquid N2 ready to create the DNA standard referement serial dilutions fir Real time test. I could help who should have these DNA.<br clear="none">
Best regards</div>
<div class="yiv2080313465gmail_quote">Il giorno 31/ott/2013 00:07, "Kwang-Poo Chang" <<a rel="nofollow" shape="rect" ymailto="mailto:kwangpoo.chang@rosalindfranklin.edu" target="_blank" href="mailto:kwangpoo.chang@rosalindfranklin.edu">kwangpoo.chang@rosalindfranklin.edu</a>> ha scritto:<br clear="none"><blockquote class="yiv2080313465gmail_quote" style="margin:0 0 0 .8ex;border-left:1px #ccc solid;padding-left:1ex;">
<div dir="ltr">Dear Carlos,<div><br clear="none"></div><div>One of my colleagues in Naples, Italy has used primer set for kinetoplast minicircle conserved region, Considering ~10,000 copies of minicircvles per Leishmania, this is probably the most sensitive PCR, although the primer set can also amply all trypanosomes or trypanosomatid protozoa (including Leptomonas reported to exist in the Indian kala-azar splenic aspirates). Specificity could be an issue in that sense.</div>
<div><br clear="none"></div><div>KP </div></div><div class="yiv2080313465gmail_extra"><br clear="none"><br clear="none"><div class="yiv2080313465gmail_quote">On Thu, Oct 10, 2013 at 7:31 PM, Carlos Lobo <span dir="ltr"><<a rel="nofollow" shape="rect" ymailto="mailto:carloshlobo@gmail.com" target="_blank" href="mailto:carloshlobo@gmail.com">carloshlobo@gmail.com</a>></span> wrote:<br clear="none">
<blockquote class="yiv2080313465gmail_quote" style="margin:0 0 0 .8ex;border-left:1px #ccc solid;padding-left:1ex;"><div dir="ltr"><div>Hello guys, good night.</div><div><span>I'm taking advantage of</span> <span>this</span> opportunity to <span>exchange </span><span>scientific</span> <span>information</span><span>,</span> <span>much</span> <span>of which I</span> <span>have learned</span> <span>to</span> <span>ask for</span> <span>help.</span><br clear="none">
<span>I'll</span> <span>start a</span> <span>project on</span> <span>canine leishmaniasis</span> <span>in northeastern</span> <span>Brazil</span> <span>and I</span> <span>need to diagnose, by</span> <span>PCR,</span> <span>dogs</span> <span>infected and</span> <span>not</span> <span>infected</span> with <span>Leish</span><span>.</span><br clear="none">
<span>I've been doing</span> <span>some analysis</span> <span>of</span> <span>PCR for</span> <span>other experiments</span><span>,</span> <span>but never</span> <span>did for</span> <span>Leish</span><span>,</span> <span>could someone give me</span> <span>some tips</span><span>? Like which primer to use? Should I collect blood or tissue samples to have more reliability?</span><br clear="none">
<span>Thank you</span><span>.</span><br clear="none"><span>Carlos Henrique</span></div></div><div><div><div class="yiv2080313465gmail_extra"><br clear="none"><br clear="none"><div class="yiv2080313465gmail_quote">2013/9/23 Tamrat Abebe Zeleke <span dir="ltr"><<a rel="nofollow" shape="rect" ymailto="mailto:tabebezeleke@gmail.com" target="_blank" href="mailto:tabebezeleke@gmail.com">tabebezeleke@gmail.com</a>></span><br clear="none">
<blockquote class="yiv2080313465gmail_quote" style="margin:0 0 0 .8ex;border-left:1px #ccc solid;padding-left:1ex;"><div dir="ltr"><div class="yiv2080313465gmail_default" style="font-family:georgia, serif;">Dear Carlos,<br clear="none"><br clear="none"></div><div class="yiv2080313465gmail_default" style="font-family:georgia, serif;">
<div>I agree with the comment given by Phillip Lawyer. The
molecular analysis of strains or isolates from India, Kenya, and South Western
Ethiopia also supports this notion. However, the issue of distinct strains in
Sudan and North Ethiopia opts for the fact that East Africa may be the origin
of at least the naughty L. donovani strains circulating in the region. </div>
</div></div><div class="yiv2080313465gmail_extra"><br clear="all"><div><div dir="ltr"><div><font face="tahoma, sans-serif"><span style="color:rgb(111,168,220);">Tamrat Abebe</span><br clear="none"></font></div><font color="#6fa8dc"><font face="tahoma, sans-serif">Addis Ababa University College of Health Sciences, School of Medicine ,<br clear="none">
Department of Microbiology, Immunology & Parasitology </font></font><div><font color="#6fa8dc"><font face="tahoma, sans-serif">Tikur Anbessa Hospital</font></font></div><div><font color="#6fa8dc"><font face="tahoma, sans-serif">Second floor room number 76<br clear="none">
Tel: <a href="" rel="nofollow" shape="rect">+251 911 447227</a>(mobile)<br clear="none">Email:</font><font face="comic sans ms, sans-serif"> </font><a rel="nofollow" shape="rect" ymailto="mailto:tamrat.abebe@aau.edu.et" target="_blank" href="mailto:tamrat.abebe@aau.edu.et"><font face="tahoma, sans-serif">tamrat.abebe@aau.edu.et</font></a></font><div>
<font color="#6fa8dc" face="comic sans ms, sans-serif"><b><br clear="none"></b></font><div><font color="#666666" face="comic sans ms, sans-serif"><b><br clear="none"></b></font> <br clear="none"><br clear="none"> <br clear="none"></div></div></div></div></div>
<br clear="none"><br clear="none"><div class="yiv2080313465gmail_quote">On Mon, Sep 16, 2013 at 4:37 AM, Lawyer, Phillip (NIH/NIAID) [E] <span dir="ltr"><<a rel="nofollow" shape="rect" ymailto="mailto:PhillipL@niaid.nih.gov" target="_blank" href="mailto:PhillipL@niaid.nih.gov">PhillipL@niaid.nih.gov</a>></span> wrote:<br clear="none">
<blockquote class="yiv2080313465gmail_quote" style="margin:0px 0px 0px 0.8ex;padding-left:1ex;border-left-color:rgb(204,204,204);border-left-width:1px;border-left-style:solid;">Dear Carlos,<br clear="none">
<br clear="none">
<br clear="none">
<br clear="none">
For what it's worth, I believe it most likely happened the other way around: Leishmania donovani was probably introduced to East Africa from India during the late 1800s when laborers were brought from India to work in the Kenya building the railroad from Mombasa to Uganda and on other infrastructure projects. Leishmania donovani in East Africa is manifest as kala azar and is anthroponotic, the same as in India. The main vector in Kenya is Phlebotmus martini, which tends to breed in termite mounds, often associated with human dwellings. Other Symphlebotomus species, Ph. vansomerenae and Ph. celiae have also been implicated in L. donovani transmission.<br clear="none">
<br clear="none">
<br clear="none">
<br clear="none">
Regards,<br clear="none">
<br clear="none">
<br clear="none">
<br clear="none">
Phil Lawyer<br clear="none">
<br clear="none">
________________________________<br clear="none">
From: Carlos Brisola Marcondes [<a rel="nofollow" shape="rect" ymailto="mailto:cbrisolamarcondes@gmail.com" target="_blank" href="mailto:cbrisolamarcondes@gmail.com">cbrisolamarcondes@gmail.com</a>]<br clear="none">
Sent: Friday, August 30, 2013 8:41 AM<br clear="none">
To: Leish-L; <a rel="nofollow" shape="rect" ymailto="mailto:leish-l-bounces@lineu.icb.usp.br" target="_blank" href="mailto:leish-l-bounces@lineu.icb.usp.br">leish-l-bounces@lineu.icb.usp.br</a><br clear="none">
Subject: [Leish-l] why is Leishmania donovani restricted to humans in Indian subcontinent?<br clear="none">
<div><div><br clear="none">
Dear all,<br clear="none">
Leishmania donovani seems to have been introduced from East Africa to Indian subcontinent, where it has infected mostly humans, differently from East African foci.<br clear="none">
Why does this occur? Is this caused by feeding preferences of Phlebotomus argentipes, which bites mostly ruminants and humans and is associated to houses? Or are dogs rarer in that region than in Brazil, where these animals are important reservoirs of Leishmania infantum and frequently bitten by Lutzomyia longipalpis?<br clear="none">
<br clear="none">
Sincerely yours<br clear="none">
prof. dr. Carlos Brisola Marcondes<br clear="none">
Dept. Microbiol. Imunol. Parasitol./CCB<br clear="none">
Federal University of Santa Catarina<br clear="none">
Florianópolis (SC)<br clear="none">
CV: <a rel="nofollow" shape="rect" target="_blank" href="http://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4783901J2">http://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4783901J2</a><br clear="none">
blog: <a rel="nofollow" shape="rect" target="_blank" href="http://entomomedica.blogspot.com.br/">http://entomomedica.blogspot.com.br/</a><br clear="none">
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