[Leish-l] why is Leishmania donovani restricted to humans inIndian subcontinent?

Σωτηριάδου Καίτη ksoteriadou at pasteur.gr
Fri Nov 1 08:34:49 BRST 2013


 

Hi! In our hands also 13A/13B kDNA had the same problems. We came to the conclusion that the balance sensitivity/specificity is not good. Specifity/contamination is an issue in that sense.

If you want to discriminate species belonging to the L. donovani complex  k26-PCR, CPB and hsp90 give the same results that are not always in agreement with MLEE. ITS (among others) is an excellent tool for discriminating species causing VL or CL in Middle East. Please note that the latter tools can be applied on clinical samples which is an important issue. I could sent you the relevant papers in a pdf format or the citation, there are several. Please let me know.

Ketty Soteriadou (Greece)

 

 
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.

Amer
Palestine



 


________________________________

From: Stefano Reale <stefano.reale at izssicilia.it>
To: Kwang-Poo Chang <kwangpoo.chang at rosalindfranklin.edu> 
Cc: "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>; Rossella Colomba Lelli <rossella.lelli at izssicilia.it>; Leish-L <Leish-L at lineu.icb.usp.br>; "caporalevincenzo at gmail.com" <caporalevincenzo at gmail.com> 
Sent: Thursday, October 31, 2013 8:19 AM
Subject: Re: [Leish-l] why is Leishmania donovani restricted to humans in Indian subcontinent?


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.
Best regards
Il giorno 31/ott/2013 00:07, "Kwang-Poo Chang" <kwangpoo.chang at rosalindfranklin.edu> ha scritto:


	Dear Carlos, 

	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.

	KP 


	On Thu, Oct 10, 2013 at 7:31 PM, Carlos Lobo <carloshlobo at gmail.com> wrote:
	

		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 <mailto: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
				Florianopolis (SC)
				CV: http://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4783901J2
				blog: http://entomomedica.blogspot.com.br/
				
				
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