Sensitivitas Metode Serologi dan Polymerase Chain Reaction untuk Mendeteksi Bean Common Mosaic Potyvirus pada Kacang Panjang

• Main Authors: Sherli Anggraini, Sri Hendrastuti Hidayat
• Format: Article
• Language: Indonesian
• Published: Perhimpunan Fitopatologi Indonesia 2014-08-01
• Series: Jurnal Fitopatologi Indonesia
• Subjects: enzym-linked immunosorbent assay, dot immunobinding assay, reverse transcription-polymerase chain reaction
• Online Access: http://journal.ipb.ac.id/index.php/jfiti/article/view/8518

<p>Mosaic disease in yard long bean is caused by <em>Bean common mosaic potyvirus </em>(BCMV) and has been reported to affect yield. Common method to detect infection of BCMV involves serological assay and <em>polymerase chain reaction </em>(PCR). The aims of this research is to assess the sensitivity of three methods, i.e. <em>Indirect Enzym-Linked Immunosorbent Assay </em>(I-ELISA), <em>Dot Immunobinding Assay </em>(DIBA), and <em>reverse transcription </em>(RT)-PCR as detection method for BCMV infection in yard long bean. Sensitivity level of the methods was evaluated by diluting plant extract and antisera for I-ELISA and DIBA, and cDNA as template in RT-PCR. Virus isolate from Cirebon was maintained in yard long bean in screenhouse and used for the assessment. Absorbance value of ELISA showed that dilution end point for I-ELISA was reached at 10^-3 and 10^-2 of plant extract and antisera dilution, respectively. Positive infection was still detected using DIBA when the plant extract was diluted up to 10^-5 based on development of color intensity on nitrocellulose membrane. Specific viral DNA fragment was still amplified when cDNA was diluted up to 10^-4, indicated higher sensitivity level of RT-PCR method.</p>