Genetic Diversity and Phylogenetic Analysis of the Iranian Leishmania Parasites Based on HSP70 Gene PCR-RFLP and Sequence Analysis

Despite the broad distribution of leishmaniasis among Iranians and animals across the country, little is known about the genetic characteristics of the causative agents. Applying both HSP70 PCR-RFLP and sequence analyses, this study aimed to evaluate the genetic diversity and phylogenetic relationships among Leishmania spp. isolated from Iranian endemic foci and available reference strains. A total of 36 Leishmania isolates from almost all districts across the country were genetically analyzed for the HSP70 gene using both PCR-RFLP and sequence analysis. The original HSP70 gene sequences were aligned along with homologous Leishmania sequences retrieved from NCBI, and subjected to the phylogenetic analysis. Basic parameters of genetic diversity were also estimated. The HSP70 PCR-RFLP presented 3 different electrophoretic patterns, with no further intraspecific variation, corresponding to 3 Leishmania species available in the country, L. tropica, L. major, and L. infantum. Phylogenetic analyses presented 5 major clades, corresponding to 5 species complexes. Iranian lineages, including L. major, L. tropica, and L. infantum, were distributed among 3 complexes L. major, L. tropica, and L. donovani. However, within the L. major and L. donovani species complexes, the HSP70 phylogeny was not able to distinguish clearly between the L. major and L. turanica isolates, and between the L. infantum, L. donovani, and L. chagasi isolates, respectively. Our results indicated that both HSP70 PCR-RFLP and sequence analyses are medically applicable tools for identification of Leishmania species in Iranian patients. However, the reduced genetic diversity of the target gene makes it inevitable that its phylogeny only resolves the major groups, namely, the species complexes.

Evaluation of Anti-Toxoplasma IgG, IgM, and IgA in Mothers with Spontaneous Abortion in Zanjan, Northwest Iran

Toxoplasma gondii is one of the major agents of infectious abortions and due to its worldwide distribution can threat healthy pregnant women who had no previous exposure to this parasite. The present study was designed to investigate the contribution of T. gondii to spontaneous abortions in Zanjan, Northwest of Iran, using ELISA method. Blood Samples were collected from 264 mothers referred to the provincial hospitals of Zanjan due to spontaneous abortion. The sera were isolated and subjected to evaluate the anti-Toxoplasma IgG, IgM and IgA antibodies. The results showed IgG positive (IgG+) in 99 cases (37.5%). A total of 68 women (25.8%) showed seroconversion with IgM or IgA or both IgM and IgA. They included: IgM+ in 21 (8.0%), IgA+ in 23 (8.7%) and both IgM+ and IgA+ in 24 (9.1%) subjects. In 23 cases, positive titers of IgM and IgG were accompanied. In general, the analysis of anti-Toxoplasma antibody patterns, showed that about 17% of the spontaneous abortions were associated with serological patterns of acute infection. According to these findings, a considerable proportion of spontaneous abortions can be attributed to T. gondii in the study area.

Frequency of HLA-DRB1 alleles in rheumatoid arthritis patients in Zahedan, southeast Iran

Analysis of the role of different alleles of human leukocyte antigen [HLA] in rheumatoid arthritis [RA] patients is necessary in many populations and geographical areas. The aim of the present study was to investigate the frequency of HLA-DRB1 alleles in RA patients, comparing with that in control group in southeast Iran. Case-control study of rheumatoid arthritis patients referred to rheumatology clinic at university hospital. The frequency of HLA-DRB1 alleles was determined in 79 RA patients and 93 healthy subjects in Zahedan, southeast Iran. HLA-DRB1 allele types were identified by polymerase chain reaction with sequence-specific primer [PCR-SSP]. The HLA-DRB1*10 allele showed a significantly higher frequency in patients with RA [OR=2.698, 95% CI=1.087-6.699, P=.045], while the frequency of DRB1*03 allele in these subjects was significantly lower than that in the control group [OR=0.447, 95% CI=0.2285-0.8729, P=.021]. The frequencies of DRB1*01, DRB1*04, DRB1*07, DRB1*09, DRB1*11, DRB1*13, DRB1*14, DRB1*15, DRB1*16 were not significantly different between RA subjects and the control group. The data suggest that the DRB1*10 allele is a risk factor and DRB1*03 is protective for RA in this population