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http://dspace.udla.edu.ec/handle/33000/10875Registro completo de metadatos
| Campo DC | Valor | Lengua/Idioma |
|---|---|---|
| dc.contributor.advisor | Calvopiña Hinojosa, Segundo Manuel | - |
| dc.creator | Proaño Montenegro, María Belén | - |
| dc.date.accessioned | 2019-04-10T21:05:31Z | - |
| dc.date.available | 2019-04-10T21:05:31Z | - |
| dc.date.issued | 2019 | - |
| dc.identifier.citation | Proaño Montenegro, M. B. (2019) Desarrollo, estandarización y aplicación de la técnica molecular PCR en tiempo real para la detección y cuantificación de ADN del trematodo Amphimerus spp en metacercarias obtenidas de peces (Tesis de pregrado). Universidad de las Américas, Quito. | es_ES |
| dc.identifier.other | UDLA-EC-TIB-2019-17 | - |
| dc.identifier.uri | http://dspace.udla.edu.ec/handle/33000/10875 | - |
| dc.description | The World Health Organization (WHO) considers trematode infections as neglected tropical diseases. In Ecuador there are 3 trematodiasis that are prevalent in tropical zones, paragonimiasis, fasciolasis and amphimeriasis Amphimeriasis was recently and for the first time described in Ecuador, considered an emerging disease, infecting people, cats and dogs in the provinces of Esmeraldas and Manabí. Amphimerus spp. is in the bile ducts of the infected causing disease. The life cycle of Amphimerus requires 2 intermediate hosts; the first ones are invertebrates (river snails) while the second ones are several species of freshwater fish. The infection is caused by the ingestion of raw or undercooked fish infected with metacercarie. The identification of metacercarie in fish is done by dissection, artificial digestion and microscopic observation; However, sensitivity, specificity and efficiency are affected by this laborious technique. For this reason, the development of molecular techniques such as real-time PCR (qPCR) is required for the rapid and specific identification of Amphimerus DNA in different fish species in endemic areas. In this research, a Real Time PCR was standardized using in house designed primers directed to the ITS2 region of Amphimerus, it was standardized by gradients of temperature of hybridization and denaturation; and variations in the concentration of primers. No statistically significant differences were found in the threshold cycle (Ct) resulting from each variation; however, the parameters selected were those that contained a lower Ct due to the higher percentage of sensitivity. The calibration curves obtained maintained an efficiency greater than 90 percent and a correlation coefficient (R2) greater than 0.98, indicating that the level of reliability of the technique performed is high. The technique identifies 1 metacercarie and a DNA concentration of 0.05 ng-μL. The specificity was determined by comparing DNA from two trematodes Paragonimus sp. and Fasciola sp., finding that the primers were specific for Amphimerus. In experiments with infected fish muscle, was determined that the weight or the amount of muscle affects the DNA detection of Amphimerus. In conclusion, the developed technique of Real Time PCR proved to be highly sensitive, specific and reliable in the detection of the trematode Amphimerus spp. in metacercarie state found in fish. This technique could be applied to differentiate and incriminate the species of fish carrying the parasite, in order to educate people at risk in the prevention of infection and take effective control-elimination measures in the community. | en |
| dc.description.abstract | La Organización Mundial de la Salud “OMS” considera a las trematodiasis como enfermedades tropicales olvidadas... | es_ES |
| dc.format.extent | 118 p. | es_ES |
| dc.language.iso | spa | es_ES |
| dc.publisher | Quito: Universidad de las Américas, 2019 | es_ES |
| dc.rights | openAccess | es_ES |
| dc.rights.uri | http://creativecommons.org/licenses/by-nc-sa/3.0/ec/ | es_ES |
| dc.subject | ENFERMEDADES INFECCIOSAS | es_ES |
| dc.subject | PRUEBAS DE ADN | es_ES |
| dc.subject | BIOTECNOLOGÍA | es_ES |
| dc.subject | PECES | es_ES |
| dc.title | Desarrollo, estandarización y aplicación de la técnica molecular PCR en tiempo real para la detección y cuantificación de ADN del trematodo Amphimerus spp en metacercarias obtenidas de peces | es_ES |
| dc.type | bachelorThesis | es_ES |
| Aparece en las colecciones: | Ingeniería en Biotecnología | |
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| Fichero | Descripción | Tamaño | Formato | |
|---|---|---|---|---|
| UDLA-EC-TIB-2019-17.pdf | 3,29 MB | Adobe PDF | Visualizar/Abrir |
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