[Leish-l] inquiry
J.-C. Dujardin
JCDujardin at itg.be
Wed Jun 8 03:38:25 BRT 2011
Dear K-P, dear all, just to inform you that together with Sanger, we
sequenced and analysed the full genome of more than 50 Ldonovani strains
(results shortly available) and that we dispose of the needed bio-informatic
platform for these diversity analyses. We are extremely interested in
increasing the representativity of our L.donovani sample (currently Bihar and
Terai).
Best regards
JC
Prof. Dr Jean-Claude Dujardin
Head of Molecular Parasitology
Instituut voor Tropische Geneeskunde
Nationalestraat, 155
B-2000 Antwerpen
Belgium
Tel. 32.3.2476358 or 355
Fax. 32.3.2476359
From: leish-l-bounces at lineu.icb.usp.br
[mailto:leish-l-bounces at lineu.icb.usp.br] On Behalf Of Chang, Kwang-Poo
Sent: dinsdag 7 juni 2011 14:03
To: Madhumita Manna; leish-l at lineu.icb.usp.br
Subject: Re: [Leish-l] inquiry
I'll be happy to make my collection of Leishmania availalbe to anyone who is
interested in setting up a 'permanent' depository of such samples.
KP
________________________________
From: Madhumita Manna [mailto:madhumita.manna09 at gmail.com]
Sent: Tue 6/7/2011 1:29 AM
To: Chang, Kwang-Poo; leish-l at lineu.icb.usp.br
Subject: Re: [Leish-l] inquiry
Dear KP,
Thank you very much for your valuable suggestions and information. I should
talk to Neeloo. Regarding the parasites, I can give you the Iist. It is in
my thesis. Unfortunately, IICB has lost most of them. Many senior Scientists
have retired who were in charge of the job.
Kind regards,
Madhumita
On Mon, Jun 6, 2011 at 11:49 PM, Chang, Kwang-Poo
<KwangPoo.Chang at rosalindfranklin.edu> wrote:
1. Neeloo singh has been doing genomic sequencing of some Indian
isolates. You may want to coordinate with her.
2. Do you have a list of Dr. Bhaduri's collection in IICB ? If so, is it
available for checking against what I have ?
3. Deep sequencing of Leishmania genomes is very doable and not terribly
expensive. The bottle-neck is the bioinformatic analyses. There is no user
friendly program for Leish genomics in so far as I know. Neeloo may have some
useful information there.
KP
________________________________
From: Madhumita Manna [mailto:madhumita.manna09 at gmail.com]
Sent: Monday, June 06, 2011 4:45 AM
To: Chang, Kwang-Poo
Subject: Re: [Leish-l] inquiry
Dear KP,
Thank you very much for your valuable suggestions. I am trying to hit as many
genetic sites are possible and we have a plan to establish a depository of
isolates of KA, PKDL for future works. Prof. Bhaduri tried hard to set up
this kind of bank and at that time we have collected and deposited a good
number of isolates in that Parasite Bank at IICB.
For entire genome sequencing, a big group needs to work together and for that
collaboration with other group is solicited.
Kind regards,
Madhumita
On Sun, Jun 5, 2011 at 11:05 PM, Chang, Kwang-Poo
<KwangPoo.Chang at rosalindfranklin.edu> wrote:
Dear Madhumitsa,
You are doing something very important. I re-iterate my suggestions as
follows:
1. Examine Leishmania grown in cultures as well as in the original samples
2. Evaluate as many genetic sites as possible, preferably the entire genomes
3. Set up a depository center for Leishmana samples
KP
________________________________
From: Madhumita Manna [mailto:madhumita.manna09 at gmail.com]
Sent: Sun 6/5/2011 7:14 AM
To: Chang, Kwang-Poo
Subject: Re: [Leish-l] inquiry
Dear KP
I have collected the isolate in the year 2010 from a patient admitted in
Calcutta National Medical College and done the RAPD, RFLP and sequencing in
recent past. The publication is underway. The work is in collaboration with
Dr. Syamal Roy, IICB and we are interested in Satluj river valley isolates as
well as we are trying to isolate more clinical isolates from other parts of
India. In this particular case, the transformed promastigotes were cultured
at Bethune College, Kolkata where we did not maintain any L.tropica strain or
UR6 that time. In 2010, we isolated five isolates collected from Kolkata of
which four patients from Bihar,one from West Bengal. So, there was no chance
of mixing in this particular case which might result occasionally due to
human errors as you told and also Prof Schnur once cautioned about the
identity of some Indian isolates long back in 1981 when he had screened 84
isolates and he apparently got one L.tropica, if I am not wrong .
I have dealt with UR6 during my Ph.D works in 1993 onwards and have seen it
close to L.tropica [about 72% homology in Jaccard's similarity index
calculated by isozyme and RAPD works]. Recently, by ITS1 RFLP, UR6 is again
found out to be matching with L.tropica.
kind regards,
Madhumita
On Sat, Jun 4, 2011 at 11:59 PM, Chang, Kwang-Poo
<KwangPoo.Chang at rosalindfranklin.edu> wrote:
It is undoubtedly very important to evaluate the genetics of all currently
existing cultured isolates in India. To be prudent , clones of these isolates
need to be included for examination as well. Over the years, we have gotten
some >300 samples through the kindness of our colleagues in different forms,
ranging from purified DNAs to live/killed parasites to clinical samples.
Sample mix-up does occur in my own lab, regardless of how meticulous we have
tried in handling them. In reality, with no disrespect to my colleagues,
samples received occasionally were not what we expected apparently due to
unintentional errors (most likely of our own in mislabeling). We subsequently
check every sample received by nagt PCR for RFLP or sequencing.
To examine all cultured isolates circulating in India is an undertaking of
great importance that can be best accomplished by a centralized facility,
like ATCC which can guarantee the identity and quality of the cultures (also
for distribution). It will be great to set up a center of such collection in
India ? I am working with Dr. Robert Modestina of ATCC to deposit some of my
collection and will be happy to provide them to any center of Leishmania
collection.
Many years ago, Sarman Singh of AIIMS analyzed a small collection of India
Leishmania cultures he gathered from different Indian colleagues and brought
to my lab, e. g. Ag83, RMRI, DD8, UR6 etc, showing their heterogeneity by
kDNA/nDNA RFLP analyses. That particular Ag83 is identical to 3 primary
cultures of L donovani Bihar isolates, whereas the UR6 is I/II allelic
hetergeneous L tropica in nagt sequence (see Table [1] of L.
odnovani/infantum complex No. 90-93 & Table [2] L tropica No. 49 in the
website:
http://rosalindfranklin.edu/dnn/portals/24/documents/microbiology/sampleList.
pdf). DNAs were all isolated from uncloned, cultured promastigotes. It is
difficult to trace the origin of the old cultures, which must have been
passed around in different labs over the years. Consequently, I can't say for
sure that the UR6 we examined is identical to yours, for example. This goes
with the information for each isolate provided as well.
Leishmania DNAs in the original sources of clinical, fly and reservoir
samples are more difficult to study, but doable. The potential problems of
using cultured promastigotes can be eliminated by that approach.
KP
________________________________
From: Madhumita Manna [mailto:madhumita.manna09 at gmail.com]
Sent: Friday, June 03, 2011 11:47 PM
To: Chang, Kwang-Poo
Cc: leish-l at lineu.icb.usp.br
Subject: Re: [Leish-l] inquiry
Dear Prof. Chang and all,
Since my Ph.D time at IICB under Prof. Bhaduri, I am trying to characterize
clinical isolates of KA, PKDL collected mainly from Bihar and very few from
West Bengal, by isozyme, RAPD methods and recently I have come across one
isolate having ITS1 sequence matching with K27. By hsp70 also, the isolate
was identified as L.tropica. The isolate was collected from a patient from
Bihar and he was suffering from KA and sensitive to SAG. This is matching
with the observations made by Sacks in 1995 taking isolates from Bihar. I
have contacted Dr. Sharma for requesting clinical isolates from his lab to
characterize them by isozymes. In order to divulge the population structure
of the causal agents for Indian KA, isolates from different regions of India
should be collected and rigorous typing involving many parameters together,
should be carried forward.
Kind regards,
Madhumita
on Sat, Jun 4, 2011 at 8:09 AM, Madhumita Manna <madhumita.manna09 at gmail.com>
wrote:
---------- Forwarded message ----------
From: Chang, Kwang-Poo <KwangPoo.Chang at rosalindfranklin.edu>
Date: Fri, Jun 3, 2011 at 11:27 PM
Subject: Re: [Leish-l] inquiry
To: "Dr.Sunil Arora" <skarora_in at yahoo.com>,
mmukhtar at tropmedicine.org, Sharman <nandlals at hotmail.com>, Petr Volf
<volf at cesnet.cz>, Hiro Goto <hgoto at usp.br>, elfadil abass
<elfadil_abass at yahoo.com>, vishwamohan_katoch at yahoo.co.in, lalit Kant ICMR
<lalitkant at icmr.org.in>
Cc: leish-l at lineu.icb.usp.br
Dear Sunil,
Glad to hear from you ! It is good for you (and anyone else) to chip
in with different views/valuable information on the subject under discussion.
I wish to say the following:
1. It is important for you and Dr. Sharma to work together,
which is very much in line with the spirit of WorldLeish.
2. Leishmania from some CL patients along the Sutluj river
valley belong to the L donovani/infantum species complex. As you pointed out,
they are refractory to in vitro cultivation and thus very different from
those in Bihar. Splenic aspirates from Shyam Sundar's VL patients produced
exuberant growth as promasigotes in 3N, as I recalled from my visit to his
clinic in the 1980's (although none of these promastigotes produces kala-azar
in hamsters). This is well-known (correct me if I should be wrong ?). The
genetic sites of these and other Bihar isolates we examined are different
from L infantum strains studied, including those from China (Xinjiang,
Beijing, Shandong, Sichuan, Gansu), Pakistan, Iran, Turkey and elsewhere (see
Waki et al. 2007 Eukaryot. Cell 6, 198). Interestingly, the Bihar isolates is
identical to one Sri Lanka CL isolate (DNA kindly provided by Y Matsumoto, U
Tokyo) in nagt sequence, consistent with published report. Most
interestingly, Shyam Sundar, JC Dujardin and their colleagues recently
reported the isolation of Leptomonas from ~7% (according to Shyam) of his
Bihar kala-alar samples. This, coupled with the difficulty of in vitro
cultures from clinical samples at least in some cases, underscores the need
to directly examine Leishmania in the original sample sources.
3. I have followed the convention of naming the causative agents
of Indian and Sudanese kala-azar/PKDL as L donovani. The gene sequences
differ between the two (we examined) as well as from all infantum isolates
(including some previously typed by others as such). However, this L
donovani/infantum species complex is the most sequence-conserved group,
consisting of only 5 nagt genetic variants with limited base substitutions
(see Fig. 6[D-4] in Waki et al.). The sequence divergence can be larger among
the infantum variants than between infantum vs donovani. Analyses of their
whole genomic sequencing may shed light on their true evolutionary
relationships in genetic terms.
4. What is the best genetic site for typing Leishmania isolates
? This is the question I have been asked repeatedly. My answer to that is try
to analyze sequence divergence in as many sites as possible, preferably the
entire genomes. Analyses of most sites can be informative down to the species
complex level. Interpretation beyond that (to the subspecies complex level,
for example) is often difficult (and thus must remain hypothetical) without a
complete database of all existing Leishmania variants. The database is
incomplete because we are unable to grow parasites from many places and there
are too many endemic sites which have not been explored.
5. With regard to the ease of isolating L infantum for
continuous cultivation from clinical, sand fly and reservoir samples,
different investigators differ in their experience. Dr. Bastien said he has
no problem. Carlos Costa also told me that he has no difficulty of growing L
chagasi from his patients in Teresina. He does have a sizable collection of
hundreds (or thousands ?) of isolates, although I was not given the actual
rate of successful isolation.
KP
________________________________
From: Dr.Sunil Arora [mailto:skarora_in at yahoo.com]
Sent: Thursday, June 02, 2011 11:55 PM
To: Chang, Kwang-Poo; mmukhtar at tropmedicine.org; Sharman; Petr Volf;
Hiro Goto; elfadil abass; vishwamohan_katoch at yahoo.co.in; lalit Kant ICMR
Cc: leish-l at lineu.icb.usp.br
Subject: Re: [Leish-l] inquiry
Dear KP and all
I have been reading the exchange of emails regarding characterisation
of leishmania isolates from Himachal in India by Dr Sharma and KP. Actually I
have also been associated with that though very briefly and had a chance to
try-n-culture some of isolates sent by Dr Sharma. The cultures became
positive but we failed to establish good passagable cultures from any of
aspirates/biopsy specimen sent by Dr Sharma, although my lab has been able to
establish some good number of primaray cultures from spleen/BM aspirates of
Ld strains in Chandigarh, although almost all the source patients belonged to
Bihar originally for these. So there is definetely some thing very different
about the leishmania strains that are circulating in that geographical area
of India. we also had a chance to use a specific PCR (that was developed in
my lab and can distinguish between an Ld and any cutaneous strain). we found
about 40-50% of isolates got Ld specific amplification while others didnot
show Ld genome. So I do believe, there is a mix of Ld and some cutaneous
strain in that area which KP is also refering too. It needs to be
investigated further... I know Dr Sharma is on to such effort soon...
regards
sunil
Dr Sunil K.Arora
Professor
In Charge HIV diagnostic and Disease Monitoring Laboratory
Department of Immunopathology
PGIMER, Chandigarh-160 012
Ph.: 0091-172-2755192(Off)
FAX: 0091-172-2744401, 2745078
Ph:+91-172-2732863 (Res); 9872866087 (cell)
email: skarora_in at yahoo.com; skarorain at gmail.com
From: "Chang, Kwang-Poo" <KwangPoo.Chang at rosalindfranklin.edu>
To: mmukhtar at tropmedicine.org; Sharman <nandlals at hotmail.com>; Petr
Volf <volf at cesnet.cz>; Hiro Goto <hgoto at usp.br>; elfadil abass
<elfadil_abass at yahoo.com>; vishwamohan_katoch at yahoo.co.in; lalit Kant ICMR
<lalitkant at icmr.org.in>
Cc: leish-l at lineu.icb.usp.br
Sent: Friday, June 3, 2011 12:03 AM
Subject: Re: [Leish-l] inquiry
Dear Maowia,
Thanks again for the info.
Are there any record available for the number of successful
isolations versus the total number attempted for a given endemic site ?
KP
From: Maowia Mukhtar [mailto: mmukhtar at tropmedicine.org ]
Sent: Wednesday, June 01, 2011 10:41 PM
To: Chang, Kwang-Poo; 'Sharman'; 'Petr Volf'; 'Hiro Goto'; 'elfadil
abass'; vishwamohan_katoch at yahoo.co.in; 'lalit Kant ICMR'
Cc: leish-l at lineu.icb.usp.br
Subject: RE: [Leish-l] inquiry
Dear KP,
Thanks for your comments and valuable notes. We have a high success
rate inc culturing Leishmania from clinical aspirates. We have even succeeded
in culturing Leishmania directly from PKDL lesions without using hamsters
although the parasite is known to be scarce in these lesions. We have
noticed that the rate of success of cultures varies between endemic regions
in Sudan as well as between the clinical forms. Sudanese Leishmania donovani
is known to be different from the Indian donovani even in their response to
antileishmania drugs and in their reaction using different diagnostic
serology tests.
Best regards
Maowia
Professor Maowia M. Mukhtar
Institute of Endemic Diseases
University of Khartoum
P.O. Box 11463, khartoum, Sudan
Mobile; +249912234268
Fax: +249183779712
From: Chang, Kwang-Poo [mailto:KwangPoo.Chang at rosalindfranklin.edu]
Sent: 02 June 2011 01:42
To: mmukhtar; Sharman; Petr Volf; Hiro Goto; elfadil abass;
vishwamohan_katoch at yahoo.co.in; lalit Kant ICMR
Cc: leish-l at lineu.icb.usp.br; Chang, Kwang-Poo
Subject: RE: [Leish-l] inquiry
Dear Maowia,
Gald to hear from you with your work-in-progress. May I ask you some
questions and share with you some info from my very limited experience with
Sudanese L donovani ? Please correct my statements, if inccorrect.
1. "difficult to maintain" means that the promastigotes emerge
after incubation of lesion aspirates/punctures in appropriate media and
replicate during successive passages, but are low in cell density and easily
lost during repetitive passages for continuous cultivation ?
2. "difficult to grow" means that the promastigotes never emerge
or emerge after incubation of lesion aspirates/punctures in appropriate
media, but subsequently fail to replicate or replicate to a limited extent on
subculture ?
3. What is the rate of success in your attempts to isolate
parasites in culture from clinical samples in each endemic site over the
years ?
4. The Sudanese strain used by many in the USA was probably
brought over decades ago by Donald Hyneman and established in Syrian Golden
hamsters by Leslie Stauber and his students. In the 70's, I had the good
fortune of working with Dennis Dwyer on his S1 or S2 strains of this origin.
As widely reported in the literature, this is an excellent model of visceral
leishmaniasis, producing typical clinical outcome consistently, including
invariable death and splenic LD loads of up to 1010 per heavily infected
spleen. I recall I had difficult time to differentiate these splenic
amastigotes into promastigotes. I did see some motile promastigotes. While a
small number remains viable, many died and appeared as ghosts during the
subsequent incubation. This has led me to suspect selection of cultivable
clones. In some, but not all of these cultures, promastigotes eventually
became 'adapted' to the culture conditions and grew, producing consistently
up to ~40 X 106 cells/ml in 4-5 days in successive passages (starting from
~106/ml = ~ 4-5 generation or replication cycles). The stationary phase
promastigotes of these cultures no longer produce good infection of hamsters,
irrespective of the passage numbers (Isolation of 'metacyclics" to produce
good infection has since been reported in the literature, if I remember
correctly). These promastigotes are more similar to the Mediterranean L
infantum than to the Indian L donovani of the Bihar region in the sequence of
their single-copy protein-coding genes, as probably already shown by many
based on sequencing other genetic sites.
KP
From: Maowia Mukhtar [mailto: mmukhtar at tropmedicine.org ]
Sent: Tuesday, May 31, 2011 2:30 PM
To: 'Sharman'; 'Petr Volf'; Chang, Kwang-Poo; 'Hiro Goto'; 'elfadil
abass'; vishwamohan_katoch at yahoo.co.in; 'lalit Kant ICMR'
Cc: leish-l at lineu.icb.usp.br
Subject: RE: [Leish-l] inquiry
Dear Colleagues,
We have very interesting experience with Sudanese L.donovani. This
group of parasite although are genetically homogenous they still cause
diverse clinical forms in Sudan ranging from VL, PKDL, ML to CL. We have
isolated L. donovani MON82 from classic cutaneous ulcers and typed them
using both isoenzyme and molecular typing techniques. Please check our
manuscript (Elamin et al. 2008. Trans R Soc Trop Med Hyg. 2008
Jan;102(1):54-7.). We have analyzed several virulence genes comparing the
isolates from the diverse clinical forms and they turned to be are highly
homologous. Immunologically they induce different immune responses (TH1/TH2).
We know the vector of L.donovani VL isolates which is P. orientalis, but we
still don't know the vector of cutaneous L. donovani parasite. We believe the
vector plays an important role in the pathogenesis and the outcome of L.
donovani infection together with the host immune response and may be some of
the parasite virulence factors. We will soon published our data on parasite
typing and host immune responses of the diverse clinical forms.
Best regards
Maowia
Professor Maowia M. Mukhtar
Institute of Endemic Diseases
University of Khartoum
P.O. Box 11463, khartoum, Sudan
Mobile; +249912234268
Fax: +249183779712
From: leish-l-bounces at lineu.icb.usp.br
[mailto:leish-l-bounces at lineu.icb.usp.br] On Behalf Of Sharman
Sent: 31 May 2011 18:13
To: Petr Volf; Chang, Kwang-Poo; Hiro Goto; elfadil abass;
vishwamohan_katoch at yahoo.co.in; lalit Kant ICMR
Cc: leish-l at lineu.icb.usp.br
Subject: Re: [Leish-l] inquiry
Hi Petr,
Thanks for enclosing the pdf. In fact the strain which we are dealing
with is not very difficult to grow but difficult to maintain, It is rK 39
positive and as I mentioned in the conversation below that even the serous
exudate from the ulcer is also positive for rK 39 dipstick. In between
because of my relocation to a different institute I was little away from this
work but I intend to resume this work in couple of months. Although in past
we tried to isolate the parasite from sand flies but did not succeed. We will
surely attempt again with the techniques described by you and others.
Contamination is the problem.
This particular focus seems to be a complex one.
NL Sharma
From: Petr Volf <mailto:volf at cesnet.cz>
Sent: Tuesday, May 31, 2011 12:36 PM
To: Chang, Kwang-Poo <mailto:KwangPoo.Chang at rosalindfranklin.edu> ;
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
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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