Immunocapture reverse transcription polymerase chain reaction for detection of sugarcane streak mosaic virus

Abstract

Detection of plant viruses can be done by protein or nucleic acid approaches. The immunocapture-reverse transcription-polymerase chain reaction (IC-RT-PCR) method is a combination of the two approaches. Research was carried out to develop and validate IC-RT- PCR based-detection method for SCSMV, which can be applied for the sugarcane seed indexing program to support the national government’s goal for sugar self-sufficiency. Evaluation of the IC-RT-PCR method was conducted using 5 field samples. Conventional PCR and serological methods, i.e. dot immunobinding assay (DIBA) and enzyme-linked immunosorbent assay (ELISA) was also performed in the same time. All field samples gave a positive reaction to SCSMV antibodies in the DIBA and ELISA methods with the intensity of the reaction varying from low to high. SCSMV was still detected on plant extract up to 104 dilution by ELISA and DIBA. Specific DNA fragments were successfully amplified from 2 field samples using the conventional PCR method; whereas the IC-RT-PCR method was successfully amplified all field samples. Optimization test showed that the IC-RT-PCR method was able to detect SCSMV from plant extract up to 10−10 dilutions. IC-RT-PCR method is more sensitive than conventional PCR and might be recommended for the indexing method to produce high-quality virus-free sugarcane seed.