Nikol Reslová

xMAP Technology: Applications in Detection of Pathogens

xMAP technology is applicable for high-throughput, multiplex and simultaneous detection of different analytes within a single complex sample. xMAP multiplex assays are currently available in various nucleic acid and immunoassay formats, enabling simultaneous detection and typing of pathogenic viruses, bacteria, parasites and fungi and also antigen or antibody interception. As an open architecture platform, the xMAP technology is beneficial to end users and therefore it is used in various pharmaceutical, clinical and research laboratories. The main aim of this review is to summarize the latest findings and applications in the field of pathogen detection using microsphere-based multiplex assays.

Molecular characterization of Toxoplasma gondii in pork meat from different production systems in the Czech Republic

Toxoplasmosis is a major public health issue, due to the presence of Toxoplasma gondii, mainly in pork. The aim of this study was to determine the occurrence of T. gondii in pigs and wild boars bred in different production systems in the Czech Republic using ELISA and qPCR methods. Our results show that T. gondii infection is widespread in pigs and wild boars bred and slaughtered in the Czech Republic and that there is a higher exposure to T. gondii in backyard slaughter operations and organic pig farming, indicating a potential risk for meat consumption. Additionally, genotyping of amplified loci for Type II suggests the presence of one clonal genotype circulating in these animals.

Molecular detection of Toxoplasma gondii in feathered game intended for human consumption in the Czech Republic

Toxoplasma gondii is an important ubiquitous protozoan parasite, which can infect almost all warm-blooded vertebrates, including humans. The diagnosis of T. gondii infection is crucial for the prevention, surveillance, and control of its transmission. Here, a triplex real-time PCR assay targeting the B1 gene and 529rep element was used to determine the presence of T. gondii in feathered game (Anas platyrhynchos and Phasianus colchicus) hunted in the Czech Republic. The prevalence of T. gondii was 5.4% in wild ducks (n = 280) and 3.4% in common pheasants (n = 350). Additionally, genotyping of 28 T. gondii-positive samples revealed the presence of archetypal genotypes II and III as well as non-archetypal genotypes combining both type II and III alleles. Our results suggest that consumption of feathered game could pose a risk of T. gondii transmission to humans in the Czech Republic.

Fast and Reliable Differentiation of Eight Trichinella Species Using a High Resolution Melting Assay

High resolution melting analysis (HRMA) is a single-tube method, which can be carried out rapidly as an additional step following real-time quantitative PCR (qPCR). The method enables the differentiation of genetic variation (down to single nucleotide polymorphisms) in amplified DNA fragments without sequencing. HRMA has previously been adopted to determine variability in the amplified genes of a number of organisms. However, only one work to date has focused on pathogenic parasites–nematodes from the genus Trichinella. In this study, we employed a qPCR-HRMA assay specifically targeting two sequential gene fragments–cytochrome c oxidase subunit I (COI) and expansion segment V (ESV), in order to differentiate 37 single L1 muscle larvae samples of eight Trichinella species. We show that qPCR-HRMA based on the mitochondrial COI gene allows differentiation between the sequences of PCR products of the same length. This simple, rapid and reliable method can be used to identify at the species level single larvae of eight Trichinella taxa.