Whatman paper [FTA Cards] for storing and transferring Leishmania DNA for PCR examination

Diagnosis of cutaneous leishmaniasis [CL] is often made based on clinical manifestation. Correct diagnosis and identification of the parasite are crucial for choosing the effective treatment and for epidemiological studies. On the other hand, determination of Leishmania species is necessary for designing appropriate control programs. Diagnosis by PCR is becoming a [gold standard]. For PCR preparation, storage and shipments of specimens are necessary. In this study, Whatman filter paper [FTA Card] was used to store and transfer samples for Leishmania identification using PCR. Among the patients who had CL lesion and referred to Parasitology Laboratory of Emam Reza Hospital, Mashhad, Iran, 44 consented cases with positive results in their direct smear were selected. An informed consent form and a questionnaire were completed and three different types of samples [direct smear, NNN culture, and spot on FTA card] were collected. DNA extraction and PCR were carried out on three different samples from each patient. PCR results using Whatman paper samples revealed a significant difference [P<0.0001] compared to the culture method but no significant difference was seen between PCR results using samples stored on Whatman paper and direct smears. The use of FTA cards is simple, rapid, and cost-effective, and can be readily employed for large-scale population screening, especially for regions where the specimens are to be transported from distant places to the laboratory

Tc1 cells percentage in patients with cutaneous leishmaniasis before and after treatment with glucantime

Various responses and different prognosis to specific treatment in different patients from one hand, and importance of IFN-gamma producer cells on the other hand impressed us to study Tc1. The study was conducted in Ghaem Medical Center and Bu-Ali Research Institute, Mashhad University of Medical Sciences, Iran from 2001 to 2002. Lymphocytes of 36 patients were counted and cultured. Percentage of different responsible immunity cells in 29 patients, were determined by Flow Cytometry System before and after medication with glucantime [IM]. Patients who showed improvement after the treatment were put into group 1 and those who did not recover were labeled group 2. In this self-control clinical trial, sampling method was consecutive non-probability and the results were analyzed by t-test consequently. The percentage of Tc1 cells showed a significant increase despite of being stimulated with Phorpol-Mristate-Acetate [PMA] among the whole studied patients and group 1 [P= 0.069 and P= 0.040, respectively]. While no significant change was observed among patients in group 2. This verifies the influence of Tc1 cells for the treatment of patients with CL and perhaps the role of glucantime in improving the cell immunity response through increasing such cells

[NK cells in cutaneous leishmaniasis and their response to medical treatment]

WHO remarked Leishmaniasis as a world wide hygienic problem with a highly considerable prevalence in Iran. Mashhad is known as one of the most important foci of the disease in our country. Different responses to medical treatment and various prognosis of patients with cutaneous leishmaniasis as well as the possible role of NK cells in prognosis prompted us to evaluate the percentage of these cells through determining CD16+56 marker by flow cytometry in different groups of patients. In this prospective study conducted in Ghaem Hospital and Bu Ali Research Institute, Mashhad University of Medical Sciences from August 2001 to October 2002, 36 patients with confirmed cutaneous Leishmaniasis were involved .Blood samples collected from them before and after treatment, 29 cases out of these patients successfully carried out the survey and 7 patients excluded for different reasons. Lymphocytes with CD16+56 markers were counted using different monoclonal antibodies such as CD16+56 +obtained from IQ products Co., the Netherlands. In this project, the percentage of NK cells in the total patients and the control group were significantly different [p=0.01] and also the similar results were obtained with patients who respond to treatment after first period of medication [p=0.02] and those who did not [p= 0.04]. Meanwhile, in our study the percentage of NK cells of the whole patients was not significantly different, before and after the treatment. In patients who were sensitive to the treatment and in those who were not the percentage of NK cells were not differing significantly as well. These results suggest that decreased number of NK cells in patient at risk of cutaneous leishmaniasis may increase the infection risk. And also suggest that Glucantime does not affect on the percentage of NK cells in the treatment of the patients