Descriptive molecular epidemiology study of Giardia duodenalis in children of Parana State, Brazil

Background and aims: We investigated the children of Parana State, Brazil the prevalence of intestinal parasitosis and the associated factors involved in the transmission of intestinal parasites, and we genotyped the Giardia duodenalis isolates obtained. Methods: Fecal samples were analyzed by established microscopic methods. G. duodenalis positive samples were subjected to genotypic characterization by PCR amplification of sequences of the glutamate dehydrogenase gene (gdh) and by enzymatic digestion with the restriction enzyme NlaIV for classification of genotypes. Results: Of the 877 samples tested, 41% were positive for some intestinal parasitosis, the most common being the presence of protozoa (87.8%). Lack of basic sanitation and poor health education were associated for the intestinal parasite cases found, and the only associated factor for giardiasis was low family income. The G. duodenalis assemblages of gdh amplified samples were 68.6% B and 31.4% AII. Conclusion: These data demonstrate the importance of epidemiological studies for the development of effective strategies with the aim of decreasing the incidence of intestinal parasites in children. Moreover, these results contribute to our knowledge of G. duodenalis assemblages circulating in the world and also offer support for future work on the molecular and clinical aspects of giardiasis

Phospholipase gene expression during Paracoccidioides brasiliensis morphological transition and infection

Phospholipase is an important virulence factor for pathogenic fungi. In this study, we demonstrate the following: (i) the Paracoccidioides brasiliensis pld gene is preferentially expressed in mycelium cells, (ii) the plb1 gene is mostly up-regulated by infection after 6 h of co-infection of MH-S cells or during BALB/c mice lung infection, (iii) during lung infection, plb1, plc and pld gene expression are significantly increased 6-48 h post-infection compared to 56 days after infection, strongly suggesting that phospholipases play a role in the early events of infection, but not during the chronic stages of pulmonary infection by P. brasiliensis

An optimized one-tube, semi-nested PCR assay for Paracoccidioides brasiliensis detection

Introduction Herein, we report a one-tube, semi-nested-polymerase chain reaction (OTsn-PCR) assay for the detection of Paracoccidioides brasiliensis. Methods We developed the OTsn-PCR assay for the detection of P. brasiliensis in clinical specimens and compared it with other PCR methods. Results The OTsn-PCR assay was positive for all clinical samples, and the detection limit was better or equivalent to the other nested or semi-nested PCR methods for P. brasiliensis detection. Conclusions The OTsn-PCR assay described in this paper has a detection limit similar to other reactions for the molecular detection of P. brasiliensis, but this approach is faster and less prone to contamination than other conventional nested or semi-nested PCR assays