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

[Kwang-Poo Chang]

Dear J-C,

Thank you so much for the pdf reprint of hsp70.

 It is great to see that the use of "clinical samples" was mentioned in the
Materials and Methods, although I didn't find further reference to them in
the Results and Discussion in that article. I may have missed ? As you
know, I feel very strongly that we ought to examine amastigotes directly in
the clinical samples instead of or in addition to cultured promastigotes. I
gather from your publications that the latter may or may not be the actual
causative agent of the disease seen in the patient of concern. We have
preliminary data, which strongly support your finding.

As I have also mentioned that ultimately we will have to do genomic
sequencing of individual amastigotes from the clinical samples. Your group
has been in active pursue of genomics, transcriptomics and proteomics in a
number of different context. You are undoubtedly knowledgeable and fully
aware of the importance of such approaches for meta-analysis.

PCR amplification of individual genes for RFLP and sequencing analysis
certainly has its merit, but this is an interim approach, facing the
difficulty of specificity versus sensitvity in selecting multicopy vs
single-copy marker genes and host-parasite sequence overlaps (when using
clinical samples). We have discussed this and other issues in our article
published in 2007 on the 1.4 kb* nagt*  that was PCR-amplified for sequnece
and RFLP analyses from hundreds of promastigote cultures obtained by
ourselves or generously providfed by colleagues mainly in the Eurasian
 continents. The outcome was verified by similar analysis of 4 addtional
single-copy genes. Still, the sample is too small in size and biased by
using cultured promastigotes, as stated in the 1st paragraph of the
Discussion section. Toward the end, we concluded that genomic sequencing
will be necessary.

During the interim period, the best approach is to PCR-amplify multiple
DNAs in different genetic sites of amastigotes direclty from clinical
samples for sequence and RFLP analysis, including the hsp70 presented in
your article.

KP


On Sat, Nov 2, 2013 at 2:28 AM, Jean-Claude Dujardin <JCDujardin at itg.be>wrote:

>  Dear KP, dear all, can I also recommend hsp70 PCR (followed by RFLP or
> sequencing)? The same primers function worldwide for all Leishmania species
> and allow discrimination of Ldonovani from Linfantum. Sensitivity in
> clinical samples is very good. See attached one of the paper of our team;
> if you want more details ask Gert Van der Auwera (in cc)
>
> Best regards
>
> JC
>
>
>
> Follow me at https://twitter.com/DujardinBiomed
>
>
>
> Prof Dr Jean-Claude Dujardin
>
> Institute of Tropical Medicine
>
> Dept of Biomedical Sciences, Head
>
> Molecular Parasitology Unit, Head
>
> Nationalestraat, 155
>
> B-2000 Antwerpen
>
> Belgium
>
>
>
> 32.3.2476355
>
>
>
> *From:* leish-l-bounces at lineu.icb.usp.br [mailto:
> leish-l-bounces at lineu.icb.usp.br] *On Behalf Of *Kwang-Poo Chang
> *Sent:* vrijdag 1 november 2013 17:59
> *To:* Σωτηριάδου Καίτη
> *Cc:* Lawyer, Phillip (NIH/NIAID) [E]; leish-l-bounces at lineu.icb.usp.br;
> Rossella Colomba Lelli; Leish-L; caporalevincenzo at gmail.com
>
> *Subject:* Re: [Leish-l] why is Leishmania donovani restricted to humans
> inIndian subcontinent?
>
>
>
> Dear Ketty,
>
>
>
> Thanks a lot for the information !
>
>
>
> Could you kindly send the pdf reprints for k26, CBP and hsp90 PCR for
> diagnostic discrimination of L donovani complex for clinical samples ? I
> assume that they work for dog, sanfly and human samples ?
>
>
>
> KP
>
>
>
> On Fri, Nov 1, 2013 at 5:34 AM, Σωτηριάδου Καίτη <ksoteriadou at pasteur.gr>
> wrote:
>
>
>
> 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
>
>
>
>
>
>
>
>
>
>
> 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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