2006, Number 3
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Rev Mex Patol Clin Med Lab 2006; 53 (3)
Molecular virology in Mexico: Improvement of SARS-Coronavirus molecular diagnosis
Charles-Niño C, Garza-Rodríguez ML, Ramos-Jiménez J, Rivas-Estilla AM
Language: Spanish
References: 15
Page: 146-150
PDF size: 68.61 Kb.
ABSTRACT
The etiological agent of SARS, an emerging infectious disease, is the coronavirus named SARS-CoV. Rapid diagnostic test are very important for case recognition and management of infected people. Because the Polymerase Chain Reaction (PCR) has a high sensitivity and specificity it has been used as a viral diagnosis tool. Several reports demonstrate SARS-CoV detection by PCR from patients during the firs week of hospitalization. The major aim of this work was to standardize and set up in our region the molecular diagnosis of this virus. We performed a cDNA synthesis from SARS CoV-RNA (kindly donated by Bernhard Notch Tropical Institute Hamburg, Germany) and simultaneously we amplified by PCR a fragment of 195 bp in a single tube. In order to increase sensibility and specificity we performed a Nested PCR to amplify a second DNA fragment. Both PCR products were detected by electrophoresis and ethidium bromide staining. Variation in amplification parameters such as, primers concentration, amplification cycles and annealing temperature, we amplified two reproducible fragments of 195 and 110 bp. Based in our results, now we can apply molecular assays, as PCR, in order to perform a highly sensitive and specific molecular diagnosis of SARS-CoV in suspected patients in our region.
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