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Pineapple mealybug wilt associated virus-2 (PMWaV-2) is an important pathogen causing Mealybug wilt of pineapple (MWP). We established a realtime quantitative RT-PCR( RT-qPCR) method with Taq Man probe based on specific primers of conserved nucleotide sequence of PMWaV-2 coat protein gene. The results showed that the method was highly sensitive to positive sample,but had no fluorescence signal to health sample and blank control. The sensitivity of RT-qPCR was about 100 times higher than general PCR. Reproducibility test revealed that the coefficients of variation between intra-and inter-assay were both within 1.85%,indicating it was a quantitative detection method for PMWaV-2 with simple operation,strong specificity,high sensitivity,and reliable reproducibility.
We established a realtime quantitative RT-PCR (RT-qPCR) method with a TaqMan probe based on specific primers of conserved nucleotide (PMWaV-2), an important pathogen causing Mealybug wilt of pineapple The results showed that the method was highly sensitive to positive sample, but had no fluorescence signal to health sample and blank control. The sensitivity of RT-qPCR was about 100 times higher than general PCR. Reproducibility test revealed that the coefficients of variation between intra-and inter-assay were both within 1.85%, indicating it was a quantitative detection method for PMWaV-2 with simple operation, strong specificity, high sensitivity, and reliable reproducibility.