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Title:	Differentiation of Species of By Means of a Test of Pcr-Rflp Based On the Coding Genes for the Sub Unit 16s Ribosomal. Diferenciación de especies de Rhodococcusmediante una prueba de PCR-RFLP basada en los genes codificantes para la subunidad 16S ribosomal
metadata.dc.creator:	Pavia, Paula Calderón, Camila Puerta PhD, Concepción
Keywords:	Rhodococcus; R. equi; R. rhodnii, PCR-RFLP.;Rhodococcus; R. equi; R. rhodnii; PCR-RFLP
Publisher:	Universidad Colegio Mayor de Cundinamarca
metadata.dc.relation:	http://hemeroteca.unad.edu.co/index.php/nova/article/view/332/1207 /*ref*/Murria P, Rosenthal K, Kobayashi G. Nocardia, Rhodococcus and related Actinomycetes. En: Medical Microbiology, Tercera edición, Editorial Mosby, USA 1998. /*ref*/Kedlaya I, Ing MB, Wong SS. Rhodococcus equi infections in immunocompetent hosts: case report and review. Clin Infect Dis 2001; 32: 39-46. /*ref*/Weinstock DM, Brown AE. Rhodococcus equi: an emerging pathogen. Clin Infect Dis 2002; 34:1379-1385. /*ref*/Prescott JF. Rhodococcus equi: an animal and human pathogen. Clin Microbiol Rev 1991; 4: 20 34. /*ref*/Hill P, Campbell JA, Petrie LA. Rhodnius prolixus and its symbiotic actinomycete: a microbiological, physiological and behavioural study. Proc R Soc Lond B, Biol Sci Sp 1976; 194: 501-525. /*ref*/Yassin AF. Rhodococcus triatomae sp. nov., isolated from a bloodsucking bug. Int J Syst Evol Microbiol 2005; 55: 1575-1579. /*ref*/Eichler S, Schaub, GA. Development of symbionts in triatomine bugs and the effects of infections with trypanosomatids. Exp Parasitol 2002; 100: 17-27. /*ref*/Beard C, Cordon-Rosales C, Durvasula R. Bacterial symbionts of the triatominae and their potential use in control of Chagas disease transmission. Annu Rev Entomol 2002; 47: 123-141. /*ref*/Beard C, Mason P, Aksoy S, Tesh R, Richards F. Transformation of an insect symbiont and expression of a foreign gene in the Chagas´ disease vector Rhodnius prolixus. Am J Trop Med Hyg 1992; 46: 195 200. /*ref*/Dotson EM, Plikaytis B, Shinnick TM, Durvasula RV, Beard CB. Transformation of Rhodococcus rhodnii, a symbiont of the Chagas disease vector Rhodnius prolixus, with integrative elements of the L1 mycobacteriophage. Infect Genet Evol 2003; 3: 103-109. /*ref*/Shoemaker SA, Fisher JH, Jones WD Jr. Scoggin CH. Restriction fragment analysis of chromosomal DNA defines different strains of Mycobacterium tuberculosis. Am Rev Respir Dis 1986; 134: 210-213. /*ref*/Sambrook JF, Russell DW. Molecular Cloning, A Laboratory Manual. Tercera edición, Cold Spring Harbor Laboratory Press, New York 2001. /*ref*/Hypsa V, Dale C. In vitro culture and phylogenetic analysis of .Candidatus Arsenophonus triatominarum. an intracellular bacterium from the triatomine bug, Triatoma infestans. Int J Syst Bacteriol 1997; 47: 1140-1144. /*ref*/Bell KS, Philp JC, Aw DW. The genus Rhodococcus. J Appl Microbiol 1998; 85: 195-210. /*ref*/Yamada H, Kobayashi M. Nitrile hydratase and its application to industrial production of acrylamide. Biosci Biotechnol Biochem 1996; 60: 1391-1400.
metadata.dc.format.*:	application/pdf
metadata.dc.type:	info:eu-repo/semantics/article info:eu-repo/semantics/publishedVersion article article article
Description:	Considering the difficulty to differentiate the species of Rhodococcus by biochemical tests, a Polymerase Chain Reaction-Restriction Fragment Length Polymorphism (PCR-RFLP) has been developed to distinguish the species of the genus Rhodococcus. R. equi, R. rhodnii and other bacterial samples were cultured on Blood agar and BHI at 26 and 37°C. The obtained DNA was amplified with the primers described by Hypsa and Dale. Products of amplification were submitted to digestion with multiple restriction enzymes and the restriction patterns were confirmed by using in silico analysis. The expected amplification fragment of 1300 pb was obtained in all tested samples. R. equi was distinguished from R. rhodnii by using endonucleases such as PstI and HindIII, and from other bacteria with endonucleases PstI, HindIII, SstI, BamHI and EcoRI. The obtained restriction patterns were confirmed by using in silico analysis. This PCR-RFLP establishes an alternative for the differentiation among species of the genus Rhodococcus such as R. equi and R. rhodnii. Dada la dificultad en diferenciar las especies de Rhodococcus por pruebas bioquímicas, se desarrolló una prueba de Reacción en Cadena de la Polimerasa (PCR), seguida de un ensayo de Polimorfismo de Longitud de Fragmentos de Restricción (PCR-RFLP) para la diferenciación de las mismas. R. equi, R. rhodnii y otras bacterias fueron cultivadas en agar sangre y BHI a 37 y 26 °C. El ADN bacteriano fue extraído y amplificado con los iniciadores descritos por Hypsa y Dale. Los productos de amplificación fueron sometidos a digestión con diversas enzimas de restricción y los patrones de restricción obtenidos fueron confirmados mediante análisis in silico. Se obtuvo el fragmento de amplificación esperado de 1300 pb en todas las bacterias analizadas. Se pudo diferenciar R. equi de R. rhodnii con las endonucleasas PstI y HindIII y con respecto a otras bacterias con PstI, HindIII, SstI, BamHI y EcoRI. Los patrones de restricción obtenidos fueron confirmados mediante análisis in silico. La prueba PCR-RFLP constituye una alternativa para la diferenciación entre especies de Rhodococcus tales como R. equi y R. rhodnii.
metadata.dc.source:	NOVA Biomedical Sciences Journal; Vol. 3, Núm. 4 (2005); 14-20 Nova; Vol. 3, Núm. 4 (2005); 14-20 NOVA Ciências Biomédicas Publicação; Vol. 3, Núm. 4 (2005); 14-20 2462-9448 1794-2470
URI:	https://repository.unad.edu.co/handle/10596/29981
Other Identifiers:	http://hemeroteca.unad.edu.co/index.php/nova/article/view/332 10.22490/24629448.332
Appears in Collections:	Revista Nova

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