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Rev. Soc. Bras. Med. Trop ; 53: e20190535, 2020. tab
Article in English | LILACS, ColecionaSUS, SES-SP | ID: biblio-1136801


Abstract Since the early 20th century, the detection of intestinal parasites has improved with the development of several techniques for parasitic structures recovery and identification, which differ in sensitivity, specificity, practicality, cost, and infrastructure demand. This study aims to review, in chronological order, the stool examination techniques and discuss their advantages, limitations, and perspectives, and to provide professionals and specialists in this field with data that lays a foundation for critical analysis on the use of such procedures. The concentration procedures that constitute the main techniques applied in routine research and in parasitological kits are a) spontaneous sedimentation; b) centrifugation-sedimentation with formalin-ethyl acetate; and c) flotation with zinc sulfate solution. While selecting a technique, one should consider the purpose of its application and the technical-operational, biological, and physicochemical factors inherent in the procedures used in stool processing, which may restrict its use. These intrinsic limitations may have undergone procedural changes driven by scientific and technological development and by development of alternative methods, which now contribute to the improvement of diagnostic accuracy.

Humans , Animals , History, 20th Century , History, 21st Century , Parasitology/history , Specimen Handling/history , Feces/parasitology , Intestinal Diseases, Parasitic/diagnosis , Parasitology/methods , Specimen Handling/methods , Sensitivity and Specificity
Rev. bras. parasitol. vet ; 27(1): 60-65, Jan.-Mar. 2018. tab
Article in English | LILACS | ID: biblio-899315


Abstract This study used several diagnostic methods to examine the occurrence of and molecularly characterize Cryptosporidium spp. in captive canaries (Serinus canaria) in southern and southeastern Brazil. A total of 498 fecal samples were purified by centrifugal-flotation using Sheather's solution. Cryptosporidium spp. diagnosis was performed using three diagnostic methods: malachite green negative staining, nested PCR targeting the 18S rRNA gene, followed by sequencing the amplified fragments, and duplex real-time PCR targeting the 18S rRNA specific to detect Cryptosporidium galli and Cryptosporidium avian genotype III. The overall positivity for Cryptosporidium spp. (total samples positive in at least one protocol) from the microscopic analysis, nested PCR and duplex real-time PCR protocol results was 13.3% (66/498). The positivity rates were 2.0% (10/498) and 4.6% (23/498) for Cryptosporidium spp. by microscopy and nested PCR, respectively. Sequencing of 20 samples amplified by nested PCR identified C. galli (3.0%; 15/498), Cryptosporidium avian genotype I (0.8%; 4/498) and Cryptosporidium avium (0.2%; 1/498). Duplex real-time PCR revealed a positivity of 7.8% (39/498) for C. galli and 2.4% (12/498) for avian genotype III. Malachite green negative staining differed significantly from nested PCR in detecting Cryptosporidium spp. Duplex real-time PCR was more sensitive than nested PCR/sequencing for detecting gastric Cryptosporidium in canaries.

Resumo Este trabalho teve como objetivos determinar a ocorrência e realizar a caracterização molecular de Cryptosporidium spp. em 498 amostras fecais de canários (Serinus canaria) criados em cativeiro, utilizando três métodos de diagnóstico: análise microscópica pela coloração negativa com verde malaquita, nested PCR seguida de sequenciamento dos fragmentos amplificados e PCR duplex em tempo real específica para detecção de Cryptosporidium galli e Cryptosporidium genótipo III de aves. A positividade total para Cryptosporidium spp. (total de amostras positivas em pelo menos um método de diagnóstico) obtida pela análise microscópica, nested PCR e PCR duplex em tempo real foi de 13,3% (66/498). As taxas de positividade para Cryptosporidium spp. foram 2,0% (10/498) e 4,6% (23/498) por microscopia e nested PCR, respectivamente. O sequenciamento de 20 amostras amplificadas pela nested PCR identificou C. galli (3,0%; 15/498), Cryptosporidium genótipo I de aves (0,8%; 4/498) e Cryptosporidium avium (0,2%; 1/498). A PCR duplex em tempo real revelou positividade de 7,8% (39/498) para C. galli e 2,4% (12/498) para Cryptosporidium genótipo III de aves. A análise microscópica diferiu significativamente da nested PCR para detecção de Cryptosporidium spp. A PCR duplex em tempo real apresentou maior sensibilidade que a nested PCR/sequenciamento para detectar as espécies/genótipos gástricos de Cryptosporidium.

Animals , Canaries/parasitology , Cryptosporidiosis/diagnosis , Cryptosporidiosis/parasitology , Cryptosporidium/isolation & purification , Brazil , DNA/analysis , Cryptosporidium/genetics , Molecular Diagnostic Techniques , Animals, Domestic
Rev. patol. trop ; 46(4): 321-330, dez. 2017. mapa, tab, graf
Article in English | LILACS | ID: biblio-913724


Intestinal parasites are among the major causative agents of diseases that affect animals and humans, especially children. In view of this, the current study evaluated the occurrence of these parasitic agents in 737 children in an urban region with excellent sanitation condition of the city of Pedreira, São Paulo, Brazil. Fecal samples from the children were processed with the use of a technique of high diagnostic efficiency (TF-Test®). The diagnosis of these samples resulted in the detection of 557 parasitic structures among eleven genera of parasites, and of 46.4% (342/737) infected children. Blastocystis spp. was found in 69.6% (238/342) of the positive samples and the monoparasitism was accompanied by symptoms in 44 children. Furthermore, 67.8% (232/342) of the infected children had close contact with pets, suggesting a possible zoonotic transmission. Lastly, this study allowed to perform health education to the children, aiming the reduction of new intestinal parasitic infections.

Humans , Child , Parasites , Child , Public Health , Diagnosis , Infections , Intestines/parasitology