Open AccessQuantitative polymerase chain reaction (Q-PCR) for detection of sugarcane streak mosaic virus
Author(s) -
Dwi Subekti,
Sri Hendrastuti Hidayat,
Tri Asmira Damayanti,
Purwono Purwono
Publication year - 2020
Publication title -
iop conference series. earth and environmental science
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.179
H-Index - 26
eISSN - 1755-1307
pISSN - 1755-1315
DOI - 10.1088/1755-1315/418/1/012062
Subject(s) - real time polymerase chain reaction , polymerase chain reaction , biology , complementary dna , virology , microbiology and biotechnology , gene , genetics
Conventional PCR is a reliable method for detecting SCSMV. The availability of quantitative PCR (qPCR) can increase the power of PCR as detection method due to its high sensitivity and ability to quantify the template DNA in the reaction mixture. The study was aimed to validate the qPCR method for detection of SCSMV, and to develop detection methods for virus indexing program in seed canes production. qPCR method developed in this study was relative-qPCR with the regression equation of Y = -7,255 * log X + 11.752 for the SCSMV standard curve, so the value of X = 10 (-0.138 * Ct + 1,620). Ct value is inversely proportional to the concentration of cDNA. The greater the Ct value, the lower the concentration of cDNA in the sample. It was proved that qPCR method is more sensitive compared to conventional PCR, because it was able to detect SCSMV in samples not detected by conventional PCR methods. The relation between the Ct value and the incidence and severity of the disease on the field was assessed using field samples for qPCR. It was shown that Ct value and virus concentration did not have relation to the incidence and severity of the disease in the fields.