ESTANDARIZACIÓN DE UNA PRUEBA DE PCR-RFLP PARA LA IDENTIFICACIÓN DE Rhodococcus rhodnii EN INSECTOS TRIATOMINOS

• Main Authors: J. Rodríguez, P. Pavía, C. Calderón
• Format: Article
• Language: English
• Published: Pontificia Universidad Javeriana 2007-12-01
• Series: Universitas Scientiarum
• Subjects: Chagas disease; Rhodococcus rhodnii; Rhodnius prolixus; symbiont bacteria; triatomine insects
• Online Access: https://docs.google.com/folder/d/0B_IdlsctkRrxVV8yVEVfRG1BQmM/edit?usp=sharing&pli=1&docId=0B_IdlsctkRrxd1BQRmg0SC1hNGs
• ISSN: 0122-7483; 2027-1352

Triatomines are insects of great importance for public health because they transmit the parasite Trypano-soma cruzi, etiological agent of Chagas disease. These insects harbour intestinal symbiotic flora from theRhodococcus genus, which provide the nutrients lacking in the insect diet, nutrients which are essentialfor normal insect development and growth. Study of these bacteria is of special interest due to thepossibility of designing paratransgenesis control strategies as well as for implications that interactionsbetween these bacteria and parasites can develop inside triatomine insects. Taking into account thedifficulty in identifying bacteria from the Rhodococcus genus by traditional methods, in this work apolymerase chain reaction (PCR) followed by a restriction fragment length polymorphism analysis wasstandardized to identify Rhodococcus rhodnii isolated from Rhodnius prolixus. Gene coding for bacterial16 S rRNA was amplified and obtained products were sequenced and digested with several restrictionenzymes. Results show a total correlation between number and size of the restriction fragments obtainedin vitro and in silico as well. Therefore, applicability of this PCR-RFLP assay for R. rhodnii identificationhas been demonstrated.