RESUMO
Disseminated microsporidiosis among HIV/AIDS patients is life-threatening. The incidence may be actually higher than what has been reported. This is due to non-specific presentations of the disseminated cases and also the insensitivity of routine diagnostic technique which contribute to delay in the treatment of the disease. In the present study, we report the use of blood specimens to detect circulating microsporidia DNA, which has not been reported for diagnosis of disseminated microsporidiosis. Blood samples from HIV/AIDSpositive patients were collected over a period of one year. These samples were subjected to PCR assay using species-specific primer EBIEF1/EBIER1. Out of 100 patients, seven were confirmed positive for E. bieneusi by PCR. A fragment of 607 bp was successfully amplified. Identification of circulating microsporidia DNA in blood samples may aid in early diagnosis, thereby allows timely administration of anti-parasitic treatment.
RESUMO
Disseminated microsporidiosis among HIV/AIDS patients is life-threatening. The incidence may be actually higher than what has been reported. This is due to non-specific presentations of the disseminated cases and also the insensitivity of routine diagnostic technique which contribute to delay in the treatment of the disease. In the present study, we report the use of blood specimens to detect circulating microsporidia DNA, which has not been reported for diagnosis of disseminated microsporidiosis. Blood samples from HIV/AIDSpositive patients were collected over a period of one year. These samples were subjected to PCR assay using species-specific primer EBIEF1/EBIER1. Out of 100 patients, seven were confirmed positive for E. bieneusi by PCR. A fragment of 607 bp was successfully amplified. Identification of circulating microsporidia DNA in blood samples may aid in early diagnosis, thereby allows timely administration of anti-parasitic treatment.
RESUMO
The protein profile of serum samples from patients with amoebic liver abscess (ALA) was compared to those of normal individuals to determine their expression levels and to identify potential surrogate disease markers. Serum samples were resolved by two dimensional electrophoresis (2-DE) followed by image analysis. The up and down-regulated protein spots were excised from the gels and analysed by MS/MS. The concentration of three clusters of proteins i.e. haptoglobin (HP), α1-antitrypsin (AAT) and transferrin in serum samples of ALA patients and healthy controls were compared using competitive ELISA. In addition, serum concentrations of HP and transferrin in samples of patients with ALA and pyogenic liver abscess (PLA) were also compared. The results of the protein 2-DE expression analysis showed that HP cluster, AAT cluster, one spot each from unknown spots no. 1 and 2 were significantly up-regulated and transferrin cluster was significantly down-regulated in ALA patients' sera (p<0.05). The MS/MS analysis identified the unknown protein spot no.1 as human transcript and haptoglobin and spot no. 2 as albumin. Competitive ELISA which compared concentrations of selected proteins in sera of ALA and healthy controls verified the up-regulated expression (p<0.05) of HP and the down-regulated expression (p<0.01) of transferrin in the former, while there was no significant difference in AAT expression (p> 0.05). However, when ALA and PLA samples were compared, competitive ELISA showed significant increased concentration of HP (p<0.05) while transferrin levels were not different. In conclusion, this study showed that HP is a potential surrogate disease marker for ALA.
