UMSAgen, método para la extracción simultánea de RNA y DNA para diagnóstico molecular / UMSAgen, the method for simultaneous extraction RNA and DNA for molecular diagnostics

INTRODUCCIÓNEl RNA y el DNA son materiales genéticos de la célula cuya extracción es útil para la medicina moderna. Actualmente existen varios métodosde extracción de RNA y DNA por separado y a costos altos. En este trabajo se describe el método UMSAgen como alternativa para laextracción simultanea de RNA total y DNA genómico para diagnósticos moleculares. MATERIAL Y MÉTODOSSe obtuvieron veinte muestras de médula ósea de pacientes con leucemia mieloide crónica. Los leucocitos fueron separados de los glóbulosrojos con tampón de lisis. Aproximadamente 3 x 106 de células fueron lisadas en guanidio y conservadas a -20 ºC hasta el momento dela extracción del RNA y DNA. Se realizaron extracciones con 3 diferentes métodos: a) El método comercial Quiagen para el RNA total, b) elmétodo clásico fenol-cloroformo para el DNA genómico y c) el método UMSAgen para extracción simultanea de RNA y DNA. RESULTADOSLa pureza del RNA total y DNA genómico extraídos por el método UMSAgen es similar a la extraída con el Kit Quiagen para el RNA y latécnica clásica para el DNA. La evaluación realizada por electroforesis en agarosa y la amplifi cación del daño molecular BCR-ABL y JAK-2V617F, por RT-PCR y PCR respectivamente, fue exitosa. CONCLUSIÓNE: método UMSAgen es una alternativa fácil y económica para la extracción de RNA total y DNA genómico en diagnóstico e investigación molecular...

INTRODUCTION The RNA and the DNA are genetic materials of the cell. The RNA/DNA extraction is necessary in modern medicine. Several RNA/DNA isolation methods exist but they are very expensive and therefore not applicable in our country. In this work, we describe the UMSAgen method as an alternative for simultaneous total RNA and genomic DNA extraction for molecular diagnosis. MATERIAL AND METHODS Twenty bone marrow samples of chronic myeloid leukemia patients were obtained. The leukocytes were separated from the erythrocytes with lyses solution; 3x106 cells were lysed in guanidio solution and conserved at -20ºC until the extraction procedure.The extraction of nucleotide acids was realized by 3 different methods: a) commercial method QuiagenTM for RNA extraction, b) classic method (phenol/chloroform) for DNA extraction and c) UMSAgen method for simultaneous RNA/DNA extraction. The two fi rst methods were used as a control test for UMSAgen.RESULTS The quality and quantity of total RNA and genomic DNA obtained by the UMSAgen method were similar to that of the QuiagenTM Kit and the classic method, respectively. The ABL-BCR and JAK-2V617F amplifi cation by RT-PCR and PCR were successful. CONCLUSIONS The UMSAgen is an easy and inexpensive altrernative method for total RNA and genomic DNA extraction in diagnostic and biomolecular research.

Prevalence of Trypanosoma cruzi infection in blood banks of seven departments of Bolivia

Trypanosoma cruzi infection was studied in 1,298 sera samples of blood banks from 7 capital departments of Bolivia, using the immunofluorescence test (IFI) and Enzyme Linked Immunosorbent Assay (Elisa). The percentages of positivity in these 7 departments have an average of 28% and are distributed as follows: Sta. Cruz 51%, Tarija 45%, Cochabamba 28%, Sucre 39%, La Paz 4.9%, Oruro 6% and Potosi 24%. The prevalence is related with the altitude levels of the different departments. However in Potosi (3,945 m) we found a 24% of prevalence, probably due to the proximity of endemic valleys to the city. The authors suggest a strict control in blood donors since there exists a great risk of infection