Soil-transmitted helminth infections remain a major public health burden in low- and middle-income countries. The traditional diagnosis by microscopic examination of fecal samples is insensitive and time-consuming. In this study, a pentaplex real-time polymerase chain reaction (PCR) was evaluated for the simultaneous detection of Ancylostoma, Necator americanus, Ascaris lumbricoides, and Strongyloides stercoralis. The results were compared with those obtained by conventional parasitological diagnostic methods. Real-time PCR was positive in 48 of 77 samples (62.3%) and microscopic examination was positive in six samples (7.8%) only (P < 0.05). In conclusion, the real-time PCR assay described in this study provides a specific and sensitive diagnostic tool for the detection of these four helminth species in epidemiological studies and monitoring of treatment programs.

A loop-mediated isothermal amplification (LAMP) assay was developed to detect Enterocytozoon bieneusi DNA for the first time from human faecal specimens. Four primers specific for Enterocytozoon bieneusi were designed corresponding to small subunit rRNA gene sequences and tested on 100 human faecal specimens. Thirty-nine of the faecal specimens (39 %) were confirmed positive for Enterocytozoon bieneusi by LAMP compared with 33 % by PCR and 32 % by light microscopy. LAMP yielded 94 % sensitivity and 88 % specificity compared with microscopy (sensitivity 48 %, specificity 76 %). No significant differences in positive detection of Enterocytozoon bieneusi were found among the three methods (P.0.05). However, LAMP has shown a substantial agreement with PCR (k50.78) and fair agreement was demonstrated between microscopy and PCR (k50.25). In conclusion, the LAMP assay proved to be useful as a simplified, rapid, sensitive and specific alternative molecular screening tool in the diagnosis of Enterocytozoon bieneusi in faecal specimens.

Abstract. This study was performed to identify circulating Plasmodium falciparum proteins in patient serum, which may be useful as diagnostic markers. Depletion of highly abundant proteins from each pooled serum sample obtained from P. falciparum-infected patients and healthy individuals was performed using the Proteoseek Antibody-Based Albumin/IgG Removal Kit (Thermo Scientific, Rockford, IL). In analysis 1, the depleted serum was analyzed directly by NanoLC-MS/MS. In analysis 2, the depleted serum was separated by two-dimensional electrophoresis followed by western blot analysis. Subsequently, the selected band was analyzed by NanoLC-MS/MS. The result of analysis 1 revealed the presence of two mature erythrocyte surface antigen (MESA) proteins and chloroquine resistance transporter protein (PfCRT). In addition, analysis 2 revealed an antigenic 75-kDa band when the membrane was probed with purified IgG from the pooled serum obtained from P. falciparum-infected patients. MS/MS analysis of this protein band revealed fragments of P. falciparum MESA proteins. Thus, in this study, two different analyses revealed the presence of Plasmodium MESA protein in pooled serum from malaria patients; thus, this protein should be further investigated to determine its usefulness as a diagnostic marker.