2 mM each dNTP, 2 5 U Taq, 2 5 μL of BSA (0 1 g/10 mL) and 1 μM f

2 mM each dNTP, 2.5 U Taq, 2.5 μL of BSA (0.1 g/10 mL) and 1 μM for each forward and reverse primer in a total of 50 μL reaction volume was used. A total of 35 cycles, each consisting of 94°C for 45 s, 59°C for 45 s, and 72°C for 1 min, were performed; XL184 supplier an initial hot start at 94°C for 3 mins and a final extension step at 72°C for 7 mins were also included. For the secondary PCR step, the PCR mixture was identical except that a concentration of 1.5 mM MgCl2 was used. A total of 40 cycles, each consisting of 94°C for 30 s, 58°C for 90 s, and 72°C for 2 mins, were performed; an initial hot start at 94°C for 3 mins and a final extension step at 72°C for 7 mins were also included.

PCR products were analyzed on 1% agarose gel and visualized by ethidium bromide

staining. The PCR products were purified using the terminator V3.1 cycle sequencing kit (Applied Biosystems, Foster, CA, USA). Sequences were assembled using the SeqMan program (DNASTAR, USA). The characteristics of study participants are presented as mean and percentage. As appropriate, Student’s t-test was used to compare the means of continuous variables, whereas categorical variables were compared using Fisher’s exact test or Pearson’s Buparlisib nmr X2 test. A logistic model was used to assess any association between potential risk factors and Cryptosporidium spp. infection; P < 0.02 according to univariate analysis was considered significant and is presented with the OR. Wald's test was used to assess the significance of variable associations. Correlations between exposure and outcome that considered possible confounding variables were evaluated by multivariate analysis by means of a logistic regression model. Only variables with P < 0.05 on Wald's test were included in the multivariate model; a backward deletion process was used. Analyses were carried out using computer software SPSS ver.12 (SPSS, USA). For both univariate and multivariate

analyses, associations were considered significant at P < 0.05. We studied 183 immunocompromised patients. Branched chain aminotransferase Their medical conditions were HIV infection in 47 (25.7%), ALL 43 (23.5%), AML 13 (7.1%), CLL 18 (9.8%), various solid cancers 22 (12%), NHL 11 (6%), post-bone marrow transplant 13 (7.1%) and post-renal transplant 16 (8.7%). One hundred and fifty one patients (82.5%) were male and 32 (17.5%) female. The majority of patients (72.7%) were over 30 years old, non-diarrheic (87%), had CD4 + T-cells counts > 100 cells/mm3 (93.4%) and were urban dwellers (76%). We considered patients had Cryptosporidium infection if their fecal samples contained typical oocysts of 4–6 μm when examined after using a modified acid-fast staining technique. We identified oocysts of Cryptosporidium in the feces of 11 of the 183 patients (6%). Demographic, environmental and clinical characteristics of the studied patients are shown in Table 1. We identified two genotypes, C.parvum and C.hominis, by 18s rRNA gene amplification, sequencing and analysis. We identified C.

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