[Leish-l] inquiry
Madhumita Manna
madhumita.manna09 at gmail.com
Tue Jun 7 03:29:50 BRT 2011
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.
>
>
>
> 1. 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.
>
>
>
> 1. 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.
>
>
>
> 1. 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.
>
>
>
> 1. 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 ?
>
>
>
> 1. “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 ?
>
>
>
> 1. What is the rate of success in your attempts to isolate parasites in
> culture from clinical samples in each endemic site over the years ?
>
>
>
> 1. 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 <volf at cesnet.cz>
>
> *Sent:* Tuesday, May 31, 2011 12:36 PM
>
> *To:* Chang, Kwang-Poo <KwangPoo.Chang at rosalindfranklin.edu> ; Sharman<nandlals at hotmail.com>; Hiro
> Goto <hgoto at usp.br> ; elfadil abass <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 <KwangPoo.Chang at rosalindfranklin.edu>
>
> *To:* Sharman <nandlals at hotmail.com> ; Hiro Goto <hgoto at usp.br> ; elfadil
> abass <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<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
> >> _______________________________________________
> >> Leish-l mailing list
> >> Leish-l at lineu.icb.usp.br
> >> http://lineu.icb.usp.br/cgi-bin/mailman/listinfo/leish-l
> >>
> >
> >
> >
> > 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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