[Leish-l] inquiry
Chang, Kwang-Poo
KwangPoo.Chang at rosalindfranklin.edu
Wed Jun 1 19:41:41 BRT 2011
Dear Petr,
Sorry for not referring to your article attached in my previous message. It is indeed a great piece of work !
In support of your observations, several CL patient isolates (Trop Med Center, Adana) and one dog isolate (Osmaniye) are referred to as L. infantum based on DNA analyses of a different genetic site in the Table for L infantum/donovani complex in the following website: http://rosalindfranklin.edu/dnn/portals/24/documents/microbiology/sampleList.pdf
The data are limited to very few isolates, which grew up in culture, out of many more patient samples, which did not grow. This observation is also consistent with your successful cultivation from only 1 patient sample of the 25 isolation attempts. Our samples were collected at an earlier time from different places in the adjacent endemic area. I am also certain we didn't use the same culture medium formulation and conditions, indicating that the CL infantum is indeed difficult to grow for isolation in culture by different groups in this area.
It is entirely possible that the other 24 non-cultivable samples may well be identical to the one you were able to grow and thus examined, but you won't know that for sure until you actually examine them all. The phenotypes of cutivability vs non-cutivability may or may not be reflected in the genetic differences of the parasites. If so, we could miss some genotypes by studying only cultivable samples unless we examine the clinical samples directly. This approach, very doable nowadays with the advance in technology, will also allow us to evaluate parasite genetic heterogeneity directly and eliminate the risk of potential selection by cultivation for genetic variants/strains/species in a given clinical sample.
You mentioned the discovery of a L infantum/donovani hybrid in another e-mail. Is this a true hybrid proven by cell cloning or actually a mixture of the two ?
KP
________________________________
From: Petr Volf [mailto:volf at cesnet.cz]
Sent: Tuesday, May 31, 2011 2:06 AM
To: Chang, Kwang-Poo; Sharman; Hiro Goto; elfadil abass
Cc: leish-l at lineu.icb.usp.br
Subject: Re: [Leish-l] inquiry
Hi 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 <mailto:KwangPoo.Chang at rosalindfranklin.edu>
To: Sharman <mailto:nandlals at hotmail.com> ; Hiro Goto <mailto:hgoto at usp.br> ; elfadil abass <mailto:elfadil_abass at yahoo.com>
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
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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
>
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