RESUMO
Blastocystis spp. is a ubiquitous protozoon in the intestinal tract of human and many animals. Microscopic examination is the main method of clinical diagnosis for Blastocystis spp., which is prone to false negative. A simple and rapid diagnosis of Blastocystis spp. infection is an important step to prevent and control blastocystosis. Here, a recombinase polymerase amplification-lateral flow dipstick (RPA-LFD) assay was developed for rapid visual detection of Blastocystis spp. DNA amplification could be performed within 18 min at 37°C. The minimum DNA detection limit was 1 pg/µL, and there was no cross-reactivity with 12 other non-target pathogens, which was consistent with the sensitivity of conventional PCR (cPCR). Furthermore, 56 fecal samples from the Third Affiliated Hospital of Xinxiang Medical University were tested using RPA and cPCR methods respectively, and the results were completely consistent. The results show that RPA-LFD method has high accuracy and visual results, which provides a new choice for the differential diagnosis and rapid field detection of Blastocystis spp.
Assuntos
Infecções por Blastocystis , Blastocystis , DNA de Protozoário , Fezes , Técnicas de Diagnóstico Molecular , Técnicas de Amplificação de Ácido Nucleico , Sensibilidade e Especificidade , Blastocystis/genética , Blastocystis/isolamento & purificação , Humanos , Infecções por Blastocystis/diagnóstico , Infecções por Blastocystis/parasitologia , Técnicas de Amplificação de Ácido Nucleico/métodos , Fezes/parasitologia , Técnicas de Diagnóstico Molecular/métodos , DNA de Protozoário/genética , Recombinases/metabolismo , Recombinases/genéticaRESUMO
Blastocystis sp. is a common parasite in the intestinal tract of humans and animals. The clinical diagnosis of Blastocystis sp. mainly depends on the microscopic observation of parasite, which can lead to false-negative results. An accurate and convenient diagnostic approach for Blastocystis sp. infection is crucial for effectively preventing and controlling blastocystosis. Herein, we developed a recombinase polymerase amplification (RPA) method for detecting Blastocystis sp. The results showed that the DNA amplification by RPA established in this study could be performed within 5 min at 37°C, with maximum band intensity observed at 30 min. The minimum detection limit of RPA was 100 fg µL−1, consistent with conventional polymerase chain reaction (cPCR). Furthermore, the RPA method exhibited no cross-reactivity with 7 other non-target pathogens in the intestinal tract. Next, the newly established RPA method was used to analyse 40 fecal samples collected clinically, and the detection results were consistent with cPCR. These results corroborate that the newly developed RPA method has good sensitivity and specificity and offers the advantage of short detection times, which can be harnessed for differential diagnosis and rapid detection of Blastocystis sp.
Assuntos
Infecções por Blastocystis , Blastocystis , Humanos , Animais , Recombinases/genética , Blastocystis/genética , Reação em Cadeia da Polimerase/métodos , Técnicas de Amplificação de Ácido Nucleico/métodos , Sensibilidade e Especificidade , Infecções por Blastocystis/diagnóstico , Reação em Cadeia da Polimerase em Tempo RealRESUMO
In order to reveal the solidification behavior of Cr in the cement clinker mineral phase, 29Si magic-angle spinning nuclear magnetic resonance, X-ray diffraction, and scanning electron microscopy with energy-dispersive X-ray spectroscopy techniques were used to analyze the morphology and composition of the cement clinker mineral phase doped with Cr. The results showed that the addition of Cr did not change the chemical environment of 29Si in the clinker mineral phase, and it was still an isolated silicon-oxygen tetrahedron. Cr affected the orientation of the silicon-oxygen tetrahedron and the coordination number of calcium, leading to the formation of defects in the crystal structure of the clinker mineral phase, by replacing Ca2+ into the mineral phase lattice to form a new mineral phase Ca3Cr2(SiO4)3. Cr acted as a stabilizer for the formation of ß-C2S in the clinker calcination. As the amount of Cr increased, the relative content of C3S decreased and the relative content of C2S increased. Further, Cr easily dissolved in C2S, while it was not found in C3S. This study is conducive to further research on the mechanism of heavy metal solidification in cement clinker. Furthermore, it is important to evaluate the environmental risk of heavy metals in the process of sludge disposal through cement kiln and promote the utilization of sludge resources and the sustainable development of the cement industry.
RESUMO
A single sequencing batch reactor (SBR) with an operating mode of anaerobic/oxic/anoxic (A/O/A) was developed to determine a simpler process to recover nitrous oxide (N2O) from synthetic wastewater containing ammonia and glucose. This SBR system was initiated in A/O mode to implement nitritation (ammonia to nitrite) and then switched to A/O/A mode. Using measurements of the dissolved N2O concentration and release rate, the total production and conversion rate of N2O were calculated to reveal the potential of producing and recovering N2O in the extended anoxic phase. Results showed that the A/O/A SBR could convert the majority of the nitrite available in the system into N2O by heterotrophic denitritation over longer anoxic periods, and a conversion rate of 77% could be achieved. As a consequence, the A/O/A SBR presents potential ability to produce and recover N2O from wastewater containing ammonia and organic carbon.