[Leish-l] primers for canine leishmaniasis

Sun Oct 20 16:42:41 BRST 2013

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/
>>>
>>>
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>>
>>
>>
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>
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