Metode DAS-ELISA dan RT-PCR untuk Deteksi Potyvirus yang Menginfeksi Bawang Merah: DAS-ELISA and RT-PCR Methods for Detection of Potyviruses Infecting Shallot | DAS-ELISA and RT-PCR Methods for Detection of Potyviruses Infecting Shallot: DAS-ELISA and RT-PCR Methods for Detection of Potyviruses Infecting Shallot

anglais. DAS-ELISA and RT-PCR Methods for Detection of Potyviruses Infecting Shallot Viral infection of shallots in Indonesia may reduce yields, mainly because it is bulb-borne. Virus detection to monitor the incidence of disease in the field or examine healthy seeds requires an accurate and reliable method. In this study, the detection of Potyvirus members, i.e., Onion yellow dwarf virus (OYDV) and Shallot yellow stripe virus (SYSV), was conducted using enzyme-linked immunosorbent assay (DAS-ELISA) and reverse transcription polymerase chain reaction (RT-PCR) from shallot leaf samples. Leaf samples were collected from several shallot growing areas in Brebes and Cirebon. Infections of OYDV and SYSV were detected using the DAS-ELISA method, then field samples that gave positive reaction were selected for further detection using the RT-PCR method, followed by nucleotide sequencing. Based on the DAS-ELISA method, OYDV and SYSV infections were detected from field samples as single and mixed infections. Potyvirus infection was confirmed by the RT-PCR method from field samples that gave a positive reaction in ELISA. Nucleotide sequence analysis showed that OYDV infection was detected in both samples that reacted positively with OYDV and SYSV antiserum. These results indicated the cross-reaction between OYDV and SYSV antisera.

indonésien. Infeksi virus pada tanaman bawang merah di Indonesia merupakan salah satu faktor yang dapat menyebabkan penurunan produktivitas tanaman, terutama karena sifatnya yang terbawa umbi.  Deteksi virus untuk memantau insidensi penyakit di lapangan atau memeriksa benih sehat membutuhkan metode yang akurat dan dapat diandalkan.  Pada penelitian ini dilakukan deteksi anggota Potyvirus, yaitu Onion yellow dwarf virus (OYDV) dan Shallot yellow stripe virus (SYSV) menggunakan metode double antibody sandwhich enzyme-linked immunosorbent assay (DAS-ELISA) dan reverse transcription polymerase chain reaction (RT-PCR) dari sampel daun bawang merah.  Sampel daun dikumpulkan dari beberapa daerah penanaman bawang merah di Brebes dan Cirebon. Infeksi OYDV dan SYSV dideteksi menggunakan metode DAS-ELISA, selanjutnya sampel lapangan yang memberikan reaksi positif dipilih untuk deteksi lebih lanjut menggunakan metode RT-PCR dan perunutan asam nukleat dengan sikuensing. Berdasarkan metode DAS-ELISA, infeksi OYDV dan SYSV berhasil dideteksi dari sampel lapangan sebagai infeksi tunggal dan infeksi campuran.  Infeksi Potyvirus terkonfirmasi dengan metode RT-PCR dari sampel lapangan yang memberikan reaksi positif dalam ELISA.  Analisis sikuen nukleotida menunjukkan infeksi OYDV baik pada sampel yang bereaksi positif dengan antiserum OYDV maupun SYSV. Hasil tersebut mengindikasikan terjadinya reaksi silang antara antiserum OYDV dan SYSV.