[Leish-l] primers for canine leishmaniasis
Profa. Adj. Cáris Maroni Nunes
caris at fmva.unesp.br
Thu Oct 31 16:24:40 BRST 2013
Dear fellows,
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.
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...
Thanks you all!
Cáris
----- Original Message -----
From: Maxy De los Santos
To: Patrick Bastien ; Carlos Lobo
Cc: Leish-L ; leish-l-bounces at lineu.icb.usp.br
Sent: Thursday, October 31, 2013 1:32 AM
Subject: Re: [Leish-l] primers for canine leishmaniasis
Dear Carlos
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.
We have used these primers with human and canine samples from cutaneous leish with excellent results.
Regards
Dr. Maxy De los Santos D
Department of Parasitology
U.S.Naval Medical Research Unit 6 (NAMRU-6)
Lima-Perú
> Date: Sun, 20 Oct 2013 20:42:41 +0200
> From: patrick.bastien at univ-montp1.fr
> To: carloshlobo at gmail.com
> CC: Leish-L at lineu.icb.usp.br; leish-l-bounces at lineu.icb.usp.br
> Subject: Re: [Leish-l] primers for canine leishmaniasis
>
> Dear Carlos,
> Have a look at three excellent papers by Lachaud et al. (JCM 2001, JCM
> 2002 and Parasitol. .2002). Although they seem old, there is a lot of
> valuable information in it, and the things have not changed much. The
> primers by Mary et al. and Chargui et al. seem to have improved this
> PCR. (Obviously, this is a personal opinion, and certainly many
> researchers can write other things)
> If you cannot have access to the one in Parasitology, I can send it to you.
> In brief, if you use a repeated nuclear DNA target, you will have a
> robust sensitivity that will enable you to detect most 'sick' animals.
> If you use the kinetoplast minicircle, you will get a
> 'hyper-sensitivity" that will allow you detecting all infected animals
> (including asymptomatic ones) but that will necessitate drastic
> working conditions (as you will be exposed to an extreme sensitivity
> to carry-over contaminations).
> Although it has been used with success, I do not advise you to use a
> nested PCR for the same reasons.
> In any case, you have to 'optimize' your PCR using first parasite
> promastigotes and then using seeded 'mimicked' samples.
> Good luck !
> and best wishes
> P. Bastien
>
> Pr Patrick Bastien
> Director, Departement de Parasitologie-Mycologie
> Centre National de Reference des Leishmanioses
> Vice-Director, "MiVEGEC" unit research UMR5290 CNRS/224 IRD/UM1/UM2
> Faculte de Medecine, Université Montpellier 1
> 39 Av. Charles Flahault
> 34295 Montpellier cedex 5, France
>
> ----- Message de carloshlobo at gmail.com ---------
> Date : Thu, 10 Oct 2013 21:31:30 -0300
> De : Carlos Lobo <carloshlobo at gmail.com>
> Répondre à : Carlos Lobo <carloshlobo at gmail.com>
> Objet : Re: [Leish-l] why is Leishmania donovani restricted to
> humans in Indian subcontinent?
> À : Tamrat Abebe Zeleke <tabebezeleke at gmail.com>
> Cc : Leish-L <Leish-L at lineu.icb.usp.br>, "Lawyer, Phillip
> (NIH/NIAID) [E]" <PhillipL at niaid.nih.gov>,
> "leish-l-bounces at lineu.icb.usp.br" <leish-l-bounces at lineu.icb.usp.br>
>
>
> > Hello guys, good night.
> > I'm taking advantage of this opportunity to exchange scientific information,
> > much of which I have learned to ask for help.
> > I'll start a project on canine leishmaniasis in northeastern Brazil
> > and I need
> > to diagnose, by PCR, dogs infected and not infected with Leish.
> > I've been doing some analysis of PCR for other experiments, but never did
> > for Leish, could someone give me some tips? Like which primer to use?
> > Should I collect blood or tissue samples to have more reliability?
> > Thank you.
> > Carlos Henrique
> >
> >
> > 2013/9/23 Tamrat Abebe Zeleke <tabebezeleke at gmail.com>
> >
> >> Dear Carlos,
> >>
> >> 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.
> >>
> >> Tamrat Abebe
> >> Addis Ababa University College of Health Sciences, School of Medicine ,
> >> Department of Microbiology, Immunology & Parasitology
> >> Tikur Anbessa Hospital
> >> Second floor room number 76
> >> Tel: +251 911 447227(mobile)
> >> Email: tamrat.abebe at aau.edu.et
> >> *
> >> *
> >> *
> >> *
> >>
> >>
> >>
> >>
> >> On Mon, Sep 16, 2013 at 4:37 AM, Lawyer, Phillip (NIH/NIAID) [E] <
> >> PhillipL at niaid.nih.gov> wrote:
> >>
> >>> Dear Carlos,
> >>>
> >>>
> >>>
> >>> 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.
> >>>
> >>>
> >>>
> >>> Regards,
> >>>
> >>>
> >>>
> >>> Phil Lawyer
> >>>
> >>> ________________________________
> >>> From: Carlos Brisola Marcondes [cbrisolamarcondes at gmail.com]
> >>> Sent: Friday, August 30, 2013 8:41 AM
> >>> To: Leish-L; leish-l-bounces at lineu.icb.usp.br
> >>> Subject: [Leish-l] why is Leishmania donovani restricted to humans in
> >>> Indian subcontinent?
> >>>
> >>> Dear all,
> >>> Leishmania donovani seems to have been introduced from East Africa to
> >>> Indian subcontinent, where it has infected mostly humans, differently from
> >>> East African foci.
> >>> 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?
> >>>
> >>> Sincerely yours
> >>> prof. dr. Carlos Brisola Marcondes
> >>> Dept. Microbiol. Imunol. Parasitol./CCB
> >>> Federal University of Santa Catarina
> >>> Florianópolis (SC)
> >>> CV: http://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4783901J2
> >>> blog: http://entomomedica.blogspot.com.br/
> >>>
> >>>
> >>> --
> >>> 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
> >>>
> >>
> >>
> >>
> >> --
> >> * 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
> >>
> >>
> >
> > --
> > This email was sent by icb.usp.br
> >
>
>
> ----- Fin du message de carloshlobo at gmail.com -----
>
>
>
> _______________________________________________
> 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
--
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/20131031/9164c453/attachment.htm>
More information about the Leish-l
mailing list