Resumo

SOTO, Marianela; GONZALEZ, Lien; PERALTA, Esther Lilia  e  DURAN-VILA, Nuria. COMPARISON OF DETECCTION LIMIT OF NUCLEIC ACID ANALYSIS METHODS FOR THE HOP STUNT VIROID (HSVd) IN CITRUS. Rev. Protección Veg. [online]. 2007, vol.22, n.2, pp. 97-103. ISSN 2224-4697.

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.

Palavras-chave : Viroid; HSVd; sPAGE; NASH; RT-PCR; molecular diagnosis.

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