COMPARISON OF DETECCTION LIMIT OF NUCLEIC ACID ANALYSIS METHODS FOR THE HOP STUNT VIROID (HSVd) IN CITRUS

Revista de Protecci贸n Vegetal

Soto, Marianela¹  Peralta, Esther Lilia⁴  González, Lien²  Duran-Vila, Nuria³ 

[1] Centro Nacional de Sanidad Agropecuaria (CENSA), La Habana, Cuba;Universidad de La Habana, Ciudad de La Habana, Cuba;Instituto Valenciano de Investigaciones Agrarias (IVIA), Valencia, España;Instituto de Investigaciones en Fruticultura Tropical (IIFT), C. Habana

关键词: Viroid;    HSVd;    sPAGE: NASH;    RT-PCR;    molecular diagnosis;   

DOI  :  

学科分类：农业科学（综合）

来源: Centro Nacional de Sanidad Agropecuaria

【 摘 要 】

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.

【 授权许可】

CC BY-NC