[Leish-l] inquiry
Jennie Blackwell
jmb37 at cam.ac.uk
Fri Jun 3 21:06:01 BRT 2011
And have you used cloned parasite isolates to decide you have a hybrid?
Jenefer M. Blackwell
TICHR, CCHR, UWA
Phone: +61 8 94897910
From my iPhone
On 03/06/2011, at 21:37, Patrick Bastien <patrick.bastien at univ-
montp1.fr> wrote:
> Dear Petr,
>
> Two comments :
> 1) I am curious to know how you define a hybrid between L. infantum
> and L. donovani, knowing that these species are quite difficult to
> differentiate by simple sequence analysis. Is it using the MLST
> method described in your paper attached ? Are the genes you are
> using single copy or duplicated ? Have you used isoenzymes for
> confirmation ?
>
> 2) Talking of such, I am always puzzled when I see efforts for
> correlating molecular and isoenzyme-based phenogramms (Fig. 4, your
> article attached). Worse, your text clearly states that the strains
> analyzed are "unambiguously distinct from the MON-1 zymodeme (...)
> of L. infantum". This is particularly risky, as it is to use
> zymodeme names (MON-...) in a phylogenetic reconstruction using
> other intrinsic (here molecular) criteria. Indeed, a zymodeme may
> include different genotypes and a genotype may theoretically cover
> several zymodemes. A zymodeme is nothing else than an "operational
> taxonomic unit" used for classification (phenetics) (see the famous
> paper by Rioux et al. attached); as such, it has no biological nor
> evolutionary signification.
> Similarly, when you write "Among accessible isolates typed using the
> MLST method, our strain is close to MON-188, the L. infantum strain
> isolated by Gramiccia (Pratlong et al., 2003)(...)", the MON-... is
> abusively assimilated to a given strain (here ISS800), whereas one
> zymodeme normally clusters several strains, that might be found
> clustered differently using your method or another method.
> Another important point is that the MON-... is defined by the
> examination of a series of 15 given iso-enzymes, but this is also
> dependent upon the technique used (and, to a lesser degree, to the
> person/research group who defines it). That is why Rioux et al.
> recommended that each identifying center use its own code (such as
> MON- for Montpellier or LON- for London). Indeed, MON-188 in
> Montpellier migh tbe different from MON-188 in another center if the
> system used is not exactly the same.
> All this may seem trivial, but there is a real danger, as it slowly
> introduces in the minds of our colleagues scientists that different
> classifications are equivalent.
> I know that I am touching on a question which can yield hours of
> debate and thousands of e-mails; so, although I am sure you will
> want to reply to this, please note that I do not want to lauch such
> a debate online !
>
> Best wishes
> Patrick
>
> Professeur Patrick Bastien
> Laboratoire de Parasitologie-Mycologie, Faculté de Médecine
> UMR MIVEGEC (CNRS 5290 - IRD 224 - Université Montpellier 1)
> Centre National de Référence des Leishmania
> CHU de Montpellier
> 39 Avenue Charles Flahault
> 34295 Montpellier Cedex 5, France
>
> Petr Volf <volf at cesnet.cz> a écrit :
>
>> Re: [Leish-l] inquiryHi K.P.,
>> in Cukurova region, Turkey, cutaneous L. infantum (now it seems
>> that we are dealing with L.donovani/L. infantum hybrid) grew very
>> poorely if we isolated them from patients. Only 1 of 25 isolations
>> was succesfull. However, the same strain (confirmed by molecular
>> methods) grew repeatedly and very well if we isolated them from
>> sandflies. It might be useful for Dr. Sharma try to get isolates
>> from sand flies. It is very laborious but very useful, in Cukurova
>> we got about dozen of isolates by this method (all identical).
>> Patients are rK39 negative, see attached paper.
>> Best wishes
>> Petr
>> ----- Original Message -----
>> From: Chang, Kwang-Poo
>> To: Sharman ; Hiro Goto ; elfadil abass
>> Cc: leish-l at lineu.icb.usp.br
>> Sent: Sunday, May 22, 2011 1:39 AM
>> Subject: Re: [Leish-l] inquiry
>>
>>
>> Dr. NL Sharma has been working on an important CL endemic area
>> along the Satluj River valley to the south of Himalaya in India
>> (Please correct me should I be wrong for anything I said here and
>> below). I had the good fortune of visiting the site several years
>> back courtesy of Dr. Sharma.
>>
>> I believe Dr. Sharma's finding is important, since the parasites
>> there are different from the familiar Indian L donovani in Bihar.
>> The parasites are refratory to in vitro cultivation. They do
>> differentiate into promastigotes and may grow a little, but can't
>> really be subcultured to establish passageable lines. This is very
>> much reminescent of L infantum in the Mediterranean area. I recall
>> Dr. Sharma has also found rK39+ dogs, if I remember correctly. If
>> so, Satluj river valley is endemic to the infantile CL.
>>
>> Analyses of several batches of basically clinical CL samples from
>> Dr. Sharma showed evidence of L infantum, but also L tropica as
>> well as a mixture of the two in one sample. This is the same
>> picture we have noted for samples from Hatay, Turkey.
>>
>> These observations make me wonder a lot about our current
>> knowledge on the clinico-epidemiology based on data collected
>> previously by analyses of cultured promastigotes from one or few
>> 'representative samples'. Nowaday, technology makes it very doable
>> to work with biological samples for Leish DNAs directly from sand
>> flies, patients and reservoir animals.
>>
>> KP
>>
>>
>>
>> ---
>> ---
>> ---
>> ---------------------------------------------------------------------
>> From: leish-l-bounces at lineu.icb.usp.br on behalf of Sharman
>> Sent: Fri 5/20/2011 11:31 PM
>> To: Hiro Goto; elfadil abass
>> Cc: leish-l at lineu.icb.usp.br
>> Subject: Re: [Leish-l] inquiry
>>
>>
>> Dear all
>> I agree with Hiro gito, We are working on a focus where the CL is
>> predominantly caused by L. donovani, and the rK 39 STRIPS GIVE
>> POSITIVE
>> RESULTS WITH SERA as well as serous exudate from the lesion. The
>> results are
>> dependent upon species.
>> NL Sharma
>>
>> --------------------------------------------------
>> From: "Hiro Goto" <hgoto at usp.br>
>> Sent: Wednesday, May 18, 2011 12:09 AM
>> To: "elfadil abass" <elfadil_abass at yahoo.com>
>> Cc: <leish-l at lineu.icb.usp.br>
>> Subject: Re: [Leish-l] inquiry
>>
>> > Dear all,
>> > In our oppinion, DAT and rK39 for those samples are not
>> indicated since
>> > these tests are produced for the diagnosis of visceral
>> leishmaniasis.
>> > In case of tegumentary leishmaniasis, it is very appropriate the
>> > observation of J.J. Shaw appointing species specificity of
>> antibody
>> > response in these cases. Titers of antibodies are in general
>> low in these
>> > cases therefore depending on the species, it may result
>> negative. We have
>> > published a review recently in Expert Rev. Anti Infect. Ther. 8
>> (4),
>> > 419?433 (2010), Current diagnosis and treatment of cutaneous and
>> > mucocutaneous leishmaniasis, where we raise this point.
>> > Hiro Goto
>> >
>> > Citando elfadil abass <elfadil_abass at yahoo.com>:
>> >
>> >> Dear all I would recommend using DAT and rK39 strip test to
>> measure
>> >> antibody
>> >> responses and to evaluate the diagnostic efficiency for both
>> tests in
>> >> such group
>> >> of patients.
>> >>
>> >> Elfadil Abass
>> >>
>> ________________________________________________________________________________
>> >> Institute of Medical Microbiology and Hospital Hygiene
>> >> Philipps University Marburg
>> >> BMFZ / Hans-Meerwein Straße 2
>> >> D-35033 Marburg, Germany
>> >>
>> ________________________________________________________________________________
>> >>
>> >>
>> >>
>> >>
>> >> ________________________________
>> >> From: Nuha Nuwayri-Salti <racha at aub.edu.lb>
>> >> To: saad saad <saad1426 at gmail.com>; "leish-l at lineu.icb.usp.br"
>> >> <leish-l at lineu.icb.usp.br>
>> >> Sent: Fri, May 13, 2011 9:39:33 AM
>> >> Subject: Re: [Leish-l] inquiry
>> >>
>> >> Dear Saad first precaution to take is to separate your samples
>> into
>> >> several
>> >> portions each (at least 5 each being no more than a few hundred
>> (200-400)
>> >> microliters). This is a necessary precaution to avoid freezing
>> and
>> >> thawing
>> >> several times the same sample which will be the case should you
>> do
>> >> different
>> >> studies at different times which is unavoidable.
>> >>
>> >> What you can do is correlate the type(ulcerated, abscess,
>> furuncle
>> >> etc.. ) the
>> >> locale, the number and age of lesions with the levels of
>> antibody in the
>> >> sera of
>> >> these patgients and also monitor cell mediated immunity with
>> leishmanin
>> >> skin
>> >> test!
>> >> I have just published (in print) an article about having
>> circulating
>> >> parasites
>> >> in some of these patients with apparently pure cutaneous
>> disease. It is
>> >> the
>> >> first paper that revealed this fact. It would be interesting to
>> confirm
>> >> or
>> >> de-confirm this fact repeating what we did.
>> >> Best wishes
>> >>
>> >> Nuha Nuwayri-Salti MD
>> >>
>> >>
>> >>
>> >> AOA
>> >> Medical Honor Society
>> >>
>> >>
>> >>
>> >>
>> >> -----Original Message-----
>> >> From: leish-l-bounces at lineu.icb.usp.br
>> >> [mailto:leish-l-bounces at lineu.icb.usp.br]
>> >> On Behalf Of saad saad
>> >> Sent: Friday, April 29, 2011 11:23 PM
>> >> To: leish-l at lineu.icb.usp.br
>> >> Subject: [Leish-l] inquiry
>> >>
>> >> Dear all
>> >> Hi. i am Saad from Saudi Arabia. i have 60 sera from positive
>> case of
>> >> CL from south west of the country. I would like to have your
>> >> recommendations to start a good research line in CL using these
>> sera.
>> >> Thanks in advance for your help
>> >> _______________________________________________
>> >> Leish-l mailing list
>> >> Leish-l at lineu.icb.usp.br
>> >> http://lineu.icb.usp.br/cgi-bin/mailman/listinfo/leish-l
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>> >> Leish-l mailing list
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>> >>
>> >
>> >
>> >
>> > Profa. Dra. Hiro Goto
>> > Laboratório de Soroepidemiologia e Imunobiologia
>> > Instituto de Medicina Tropical de São Paulo, USP
>> > Av. Dr. Enéas de Carvalho Aguiar, 470, prédio II, quarto andar
>> > 05403-000 - São Paulo, SP
>> > Tel. +55-11-3061 7023, 3061 7056 ou 3061 7027
>> > Fax. +55-11-3061-8270
>> >
>> > _______________________________________________
>> > Leish-l mailing list
>> > Leish-l at lineu.icb.usp.br
>> > http://lineu.icb.usp.br/cgi-bin/mailman/listinfo/leish-l
>> >
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>>
>>
>> ---
>> ---
>> ---
>> ---------------------------------------------------------------------
>>
>>
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>
>
> <P.Volf_CL-infantum_Ptobbi Turkey.pdf>
> <6140_RIOUX_1990.pdf>
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