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<DIV><FONT size=2 face=Arial>Dear fellows,</FONT></DIV>
<DIV><FONT size=2 face=Arial>I also am very pleased to be part of this list and
have the opportunity to learn. I´ve been working with PCR for canine
leishmaniasis since 2000 and I´ve "suffered" of most of the problems you´ve
said and I do recomend be careful with the kinetoplast DNA
fragment contamination.</FONT></DIV>
<DIV><FONT size=2 face=Arial>I´d like to ask if you can give me any hints
on Leishmania differentiation by any PCR technique. I´ve tried using SSU rDNA,
ITS-1, kDNA, G6PD, SL mini-exon (some with RFLP, degenerated primers,
nested-PCR, multiplex) and when I have biological samples with quite few
Leishmania it seems that nothing works...</FONT></DIV>
<DIV><FONT size=2 face=Arial>Thanks you all!</FONT></DIV>
<DIV><FONT size=2 face=Arial>Cáris</FONT></DIV>
<DIV><FONT size=2 face=Arial></FONT> </DIV>
<DIV> </DIV>
<DIV>----- Original Message ----- </DIV>
<BLOCKQUOTE
style="BORDER-LEFT: #000000 2px solid; PADDING-LEFT: 5px; PADDING-RIGHT: 0px; MARGIN-LEFT: 5px; MARGIN-RIGHT: 0px">
<DIV
style="FONT: 10pt arial; BACKGROUND: #e4e4e4; font-color: black"><B>From:</B>
<A title=dmaxy@hotmail.com href="mailto:dmaxy@hotmail.com">Maxy De los
Santos</A> </DIV>
<DIV style="FONT: 10pt arial"><B>To:</B> <A
title=patrick.bastien@univ-montp1.fr
href="mailto:patrick.bastien@univ-montp1.fr">Patrick Bastien</A> ; <A
title=carloshlobo@gmail.com href="mailto:carloshlobo@gmail.com">Carlos
Lobo</A> </DIV>
<DIV style="FONT: 10pt arial"><B>Cc:</B> <A title=leish-l@lineu.icb.usp.br
href="mailto:leish-l@lineu.icb.usp.br">Leish-L</A> ; <A
title=leish-l-bounces@lineu.icb.usp.br
href="mailto:leish-l-bounces@lineu.icb.usp.br">leish-l-bounces@lineu.icb.usp.br</A>
</DIV>
<DIV style="FONT: 10pt arial"><B>Sent:</B> Thursday, October 31, 2013 1:32
AM</DIV>
<DIV style="FONT: 10pt arial"><B>Subject:</B> Re: [Leish-l] primers for canine
leishmaniasis</DIV>
<DIV><FONT size=2 face=Arial></FONT><BR></DIV>
<DIV dir=ltr>Dear Carlos<BR> <BR>If you want to detect canine
leish caused for Viannia subgenus, the primers designed by
Lopez M, Inga R, Cangalaya M, Echevarria J, Llanos-Cuentas A, Orrego C,
Arevalo J. Diagnosis of Leishmania using the polymerase chain reaction: a
simplified procedure for field work. Am J Trop Med Hyg. 1993 Sep;49(3):348-56.
PubMed PMID: 8396860. are better than others.<BR>We have used these
primers with human and canine samples from cutaneous leish with excellent
results.<BR> <BR>Regards<BR> <BR>Dr. Maxy De los Santos
D<BR>Department of Parasitology<BR>U.S.Naval Medical Research Unit 6
(NAMRU-6)<BR>Lima-Perú<BR><BR> <BR>
<DIV>> Date: Sun, 20 Oct 2013 20:42:41 +0200<BR>> From:
patrick.bastien@univ-montp1.fr<BR>> To: carloshlobo@gmail.com<BR>> CC:
Leish-L@lineu.icb.usp.br; leish-l-bounces@lineu.icb.usp.br<BR>> Subject:
Re: [Leish-l] primers for canine leishmaniasis<BR>> <BR>> Dear
Carlos,<BR>> Have a look at three excellent papers by Lachaud et al. (JCM
2001, JCM <BR>> 2002 and Parasitol. .2002). Although they seem old, there
is a lot of <BR>> valuable information in it, and the things have not
changed much. The <BR>> primers by Mary et al. and Chargui et al. seem to
have improved this <BR>> PCR. (Obviously, this is a personal opinion, and
certainly many <BR>> researchers can write other things)<BR>> If you
cannot have access to the one in Parasitology, I can send it to you.<BR>>
In brief, if you use a repeated nuclear DNA target, you will have a <BR>>
robust sensitivity that will enable you to detect most 'sick' animals.
<BR>> If you use the kinetoplast minicircle, you will get a <BR>>
'hyper-sensitivity" that will allow you detecting all infected animals
<BR>> (including asymptomatic ones) but that will necessitate drastic
<BR>> working conditions (as you will be exposed to an extreme sensitivity
<BR>> to carry-over contaminations).<BR>> Although it has been used with
success, I do not advise you to use a <BR>> nested PCR for the same
reasons.<BR>> In any case, you have to 'optimize' your PCR using first
parasite <BR>> promastigotes and then using seeded 'mimicked'
samples.<BR>> Good luck !<BR>> and best wishes<BR>> P.
Bastien<BR>> <BR>> Pr Patrick Bastien<BR>> Director, Departement de
Parasitologie-Mycologie<BR>> Centre National de Reference des
Leishmanioses<BR>> Vice-Director, "MiVEGEC" unit research UMR5290 CNRS/224
IRD/UM1/UM2<BR>> Faculte de Medecine, Université Montpellier 1<BR>> 39
Av. Charles Flahault<BR>> 34295 Montpellier cedex 5, France<BR>>
<BR>> ----- Message de carloshlobo@gmail.com ---------<BR>> Date :
Thu, 10 Oct 2013 21:31:30 -0300<BR>> De : Carlos Lobo
<carloshlobo@gmail.com><BR>> Répondre à : Carlos Lobo
<carloshlobo@gmail.com><BR>> Objet : Re: [Leish-l] why is
Leishmania donovani restricted to <BR>> humans in Indian
subcontinent?<BR>> À : Tamrat Abebe Zeleke
<tabebezeleke@gmail.com><BR>> Cc : Leish-L
<Leish-L@lineu.icb.usp.br>, "Lawyer, Phillip <BR>> (NIH/NIAID) [E]"
<PhillipL@niaid.nih.gov>, <BR>> "leish-l-bounces@lineu.icb.usp.br"
<leish-l-bounces@lineu.icb.usp.br><BR>> <BR>> <BR>> > Hello
guys, good night.<BR>> > I'm taking advantage of this opportunity to
exchange scientific information,<BR>> > much of which I have learned to
ask for help.<BR>> > I'll start a project on canine leishmaniasis in
northeastern Brazil <BR>> > and I need<BR>> > to diagnose, by PCR,
dogs infected and not infected with Leish.<BR>> > I've been doing some
analysis of PCR for other experiments, but never did<BR>> > for Leish,
could someone give me some tips? Like which primer to use?<BR>> > Should
I collect blood or tissue samples to have more reliability?<BR>> > Thank
you.<BR>> > Carlos Henrique<BR>> ><BR>> ><BR>> >
2013/9/23 Tamrat Abebe Zeleke <tabebezeleke@gmail.com><BR>>
><BR>> >> Dear Carlos,<BR>> >><BR>> >> I agree
with the comment given by Phillip Lawyer. The molecular analysis<BR>>
>> of strains or isolates from India, Kenya, and South Western Ethiopia
also<BR>> >> supports this notion. However, the issue of distinct
strains in Sudan and<BR>> >> North Ethiopia opts for the fact that
East Africa may be the origin of at<BR>> >> least the naughty L.
donovani strains circulating in the region.<BR>> >><BR>> >>
Tamrat Abebe<BR>> >> Addis Ababa University College of Health
Sciences, School of Medicine ,<BR>> >> Department of Microbiology,
Immunology & Parasitology<BR>> >> Tikur Anbessa Hospital<BR>>
>> Second floor room number 76<BR>> >> Tel: +251 911
447227(mobile)<BR>> >> Email: tamrat.abebe@aau.edu.et<BR>>
>> *<BR>> >> *<BR>> >> *<BR>> >> *<BR>>
>><BR>> >><BR>> >><BR>> >><BR>> >>
On Mon, Sep 16, 2013 at 4:37 AM, Lawyer, Phillip (NIH/NIAID) [E] <<BR>>
>> PhillipL@niaid.nih.gov> wrote:<BR>> >><BR>>
>>> Dear Carlos,<BR>> >>><BR>> >>><BR>>
>>><BR>> >>> For what it's worth, I believe it most
likely happened the other way<BR>> >>> around: Leishmania donovani
was probably introduced to East Africa from<BR>> >>> India during
the late 1800s when laborers were brought from India to work<BR>>
>>> in the Kenya building the railroad from Mombasa to Uganda and on
other<BR>> >>> infrastructure projects. Leishmania donovani in
East Africa is manifest as<BR>> >>> kala azar and is
anthroponotic, the same as in India. The main vector in<BR>> >>>
Kenya is Phlebotmus martini, which tends to breed in termite mounds,
often<BR>> >>> associated with human dwellings. Other
Symphlebotomus species, Ph.<BR>> >>> vansomerenae and Ph. celiae
have also been implicated in L. donovani<BR>> >>>
transmission.<BR>> >>><BR>> >>><BR>>
>>><BR>> >>> Regards,<BR>> >>><BR>>
>>><BR>> >>><BR>> >>> Phil Lawyer<BR>>
>>><BR>> >>> ________________________________<BR>>
>>> From: Carlos Brisola Marcondes
[cbrisolamarcondes@gmail.com]<BR>> >>> Sent: Friday, August 30,
2013 8:41 AM<BR>> >>> To: Leish-L;
leish-l-bounces@lineu.icb.usp.br<BR>> >>> Subject: [Leish-l] why
is Leishmania donovani restricted to humans in<BR>> >>> Indian
subcontinent?<BR>> >>><BR>> >>> Dear all,<BR>>
>>> Leishmania donovani seems to have been introduced from East
Africa to<BR>> >>> Indian subcontinent, where it has infected
mostly humans, differently from<BR>> >>> East African
foci.<BR>> >>> Why does this occur? Is this caused by feeding
preferences of<BR>> >>> Phlebotomus argentipes, which bites mostly
ruminants and humans and is<BR>> >>> associated to houses? Or are
dogs rarer in that region than in Brazil,<BR>> >>> where these
animals are important reservoirs of Leishmania infantum and<BR>>
>>> frequently bitten by Lutzomyia longipalpis?<BR>>
>>><BR>> >>> Sincerely yours<BR>> >>> prof.
dr. Carlos Brisola Marcondes<BR>> >>> Dept. Microbiol. Imunol.
Parasitol./CCB<BR>> >>> Federal University of Santa
Catarina<BR>> >>> Florianópolis (SC)<BR>> >>> CV:
http://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4783901J2<BR>>
>>> blog: http://entomomedica.blogspot.com.br/<BR>>
>>><BR>> >>><BR>> >>> --<BR>> >>>
This email was sent by icb.usp.br ­­<BR>> >>>
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>>><BR>> >>> --<BR>> >>><BR>> >>>
This email was sent by icb.usp.br<BR>> >>><BR>>
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