Development and assessment of loop-mediated isothermal amplification [LAMP] assay for the diagnosis of human visceral leishmaniasis in Iran

Parasitological methods for the diagnosis of Visceral leishmaniasis [VL] require invasive procedures, so serological and molecular approaches have been developed but are not generally applicable in the field. We evaluated a loop mediated isothermal amplification [LAMP] assay using blood from VL patients and compared it to nested PCR. Forty-seven subjects with clinical features [fever, hepatosplenomegaly and anemia] were confirmed positive for VL by the direct agglutination test [DAT] at titers >3200. Forty DAT negative individuals from non-endemic areas with no clinical signs or symptoms of VL served as controls. A LAMP assay was performed using a set of six primers targeting leishmania infantum kinetoplast DNA [kDNA] minicircle gene under isothermal [64 degree C] conditions. For nested PCR we used primers targeting the kDNA minicircle gene. The LAMP assay provided a detection limit of 1 parasite in 1 ml of j peripheral blood and detected L. infantum DNA in 44 of 47 DAT-confirmed VL cases, with diagnostic sensitivity of 93.6% [95% CI]. No L. infantum DNA was amplified in controls, indicating a specificity of 100%. The nested PCR yielded sensitivity of 96% [95% CI] and a specificity of 100% [95% CI]. The LAMP assay gave results similar to those of nested PCR but in a shorter time. The LAMP method is simple; requires no sophisticated equipment; has a short reaction time; and results, indicated by turbidity of the reaction mixture, are observable with the naked eye

[Study on the association of Toxocariasis with allergic rhinitis in individuals referred to Rasoul Akram Hospital, Tehran]

Toxocariasis is a common worldwide zoonotic parasite infection caused by the larvae of Toxocara catti and Toxocara canis. Allergic rhinitis is the most common chronic diseases in the upper respiratory tract. The main symptoms are sneezing, watery rhinorrhea, itching in the nose, eyes and palate. The purpose of this study was to investigate the association between toxocara seropositivity and allergic rhinitis compared with the control population. This cross-sectional study was carried out from September 2009 to February 2011 on 93 patients with allergic rhinitis and 87 control subjects. Confirmation of the diagnosis of allergic rhinitis was defined by history and positive epicutaneous prick test. Control subjects were healthy based on history and no signs of allergic rhinitis and other allergic diseases were seen. Blood and fecal samples were taken from both groups. Sera were separated, labeled and stored at -20°C until used. Stool samples were examined by a wet mount and formalin-ethyl acetate concentration technique. The diagnosis of toxocariasis was established by IgG anti Toxocara and IgE total by ELISA method. In case group [allergic rhinitis] from 93 patients, 50 patients [53.8%] were males and 43 [46.2%] were female. In the control group of 87 individuals studied, 56 [64.4%] were males and 31 [35.6%] were female. In cases and controls, 5 [5.4%] and 3 [3.4%] of sera were positive for IgG Toxocara, respectively. There was no statistical difference in Toxocara seropositivity in both groups [p =0.39]. It seems to be in contrast to worms and allergies several factors, including phase worm infections [acute and chronic], parasite load, parasite species and resistance genes are involved and this require further studies in different ages and populations

[Evaluation of serological diagnosis indirect immunofluorescence and ELISA in comparison to direct examination and culture in vulvovaginal candidiasis]

Vulvovaginal candidiasis occurs due to the overgrowth of candida in genital system mucosa of females. Symptoms and signs of vulvovaginal candidiasis are unspecific, therefore diagnosis is not certain. The aim of this research was comparison of the result of indirect immunofluorescence and ELISA with culture and direct microscopy examination in patients with vulvovaginal candidiasis. This was a comparative-descriptive study performed on 87 patients and 50 normal cases as controls. All specimens were examined using direct microscopy, culturing and complimentary test to differentiate the candida species from each other. Serological tests such as indirect immunofluorescence and ELISA were performed on sera of the patients. To compare the quantitative and qualitative data, t-student test, chi-square and exact fisher test were used, if necessary. Out of 87 specimens, 50 cases were diagnosed as vulvovaginal candidiasis. The most frequent isolated pathogens were C.albicans, C.glabrata, C.kefyr, C.inconspicua and C.famata, respectively. Also, in control group, the most frequent pathogens were C.albicans, C.glabrata and C.kefyr, respectively. In this study all of normal cases were negative in indirect immunofluorescence test and in patients group 42 person [84%] were positive and 8 [16%] negative. Control participants were negative in ELISA and in patient group 40 person [80%] were positive and 10 person [20%] negative. It seems in cases that direct microscopic and culturing methods is impossible, ELISA and indirect immunofluorescence can be used as an alternative method

[Investigation of serum C-reactive protein and mannose binding lectin levels in patients with dermatophytosis]

Dermatophytosis is common cutaneous fungal disease with worldwide distribution. Interleukin8 [IL-8] realized from keratinocytes in the presence of dermatophytic antigens causes induction of acute responses in dermatophyte infection and subsequently production of acute phase proteins occurs in hepatocytes. C-reactive protein [CRP] and Mannose binding lectin [MBL] are acute phase proteins. Since few researches in the case of acute phase proteins in dermatophytic infections has been accomplished, this study has been designed for determining serum CRP and MBL levels in patients affected to dermatophytosis. This was a cross sectional study and samples were carried out on 96 healthy individuals and 105 patients affected to dermatophytosis with non probable and in access procedure. For isolation and identification of dermatophyte direct microscopic examination, culturing and complementary examinations were done and for determination of serum CRP and MBL levels in healthy individuals and in patients ELISA test were used. For investigation of relevance between variables, Chi-square, Fisher exact, Mann-Whitney and Roc curve analysis were used and p<0.05 was considered as meaningful level. The median serum CRP level in healthy individuals and in patients group was 3.31 +/- 3.32 micro g/ml and 16.60 +/- 35.96 micro g/ml [p<0.001] respectively and the median serum MBL level was 1.53 +/- 1.87 micro g/ml and 1.97 +/- 2.03 micro g/ml [p=0.039] respectively. CRP [p<0.001] and MBL [p=0.042] were determined meaningful parameters for dermatophytosis. MBL deficiency [MBL concentrations<1 micro g/ml] was higher in control subjects [56.2%] than in patients [41.0%]. Findings of this study indicate increased concentrations of CRP and MBL in patients affected to dermatophytosis and their role in this infection. Probably observation of high frequency of MBL deficiency in healthy individuals in compare with patients group indicates that it is not predisposing factor in affecting to dermatophytosis