COMPARACIÓN DEL LÍMITE DE DETECCIÓN DE MÉTODOS DE ANÁLISIS DE ÁCIDOS NUCLEICOS PARA EL VIROIDE DEL ENANISMO DEL LÚPULO (HSVd) EN CÍTRICOS
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The detection limit of sequence polyacrilamide gel electrophoresis (sPAGE), nucleic acid hybridization (NASH) and reverse transcription-polymerase chain reaction (RT-PCR) was determined for the diagnosis of the hop stunt viroid (HSVd) in citrus hosts, using citron and cucumber plants as biological amplifiers. The detection limit of sPAGE was higher (5x10-3) when cucumber plants were analyzed than that using citron plants (5x10-2); however, it was 50 times less sensitive than that of NASH (10-4) by comparison. The RT-PCR detection limit for cucumber host was the highest (10-6) of all methods analyzed. This value was higher when the RT-PCR products were detected by NASH and no with agarose gel electrophoresis instead. RT-PCR was the most sensitive method and its sensitivity was increased when it was combined with NASH. However, the presence of contaminations associated with the high sensitivity of RT-PCR is a routine diagnosis problem. On the other hand, NASH is a method with a proper sensitivity under the conditions analyzed since the detection limit was higher than that of sPAGE, although slower than that of RT-PCR.