Comparison of IC-RT-PCR, Dot-ELISA and Indirect-ELISA for the Detection of Sorghum Mosaic Virus in Field-Grown Sugarcane Plants

Chen, Hai; Ali, Niyaz; Lv, Wenzhu; Shen, Yanan; Zhen, Qing; Lin, Yinfu; Chen, Baoshan; Wen, Ronghui

doi:10.1007/s12355-019-00738-5

Cited by 4 publications

(4 citation statements)

References 24 publications

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“…The ELISA experiment was explored by using a slightly modified procedure [ 26 ]. Plant sap from infected sugarcane, healthy sugarcane, and a definite negative sample (as a negative control) was extracted by fully grinding 1 g of blades in liquid nitrogen followed by suspension in 500 μL of PBS buffer (pH = 7.4).…”

Section: Methodsmentioning

confidence: 99%

A Rapid, Fluorescence Switch-On Biosensor for Early Diagnosis of Sorghum Mosaic Virus

et al. 2022

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For the first time, a nanobiosensor was established for Sorghum mosaic virus (SrMV) detection. The biosensor consists of cadmium telluride quantum dots (CdTe QDs) conjugated to the specific antibody (Ab) against SrMV coat protein (CP) and carbon quantum dots (C QDs) labeled with SrMV coat protein. The formation of the fluorophore-quencher immunocomplex CdTe QDs-Ab+C QDs-CP led to a distinct decrease in the fluorescence intensity of CdTe QDs. Conversely, the emission intensity of CdTe QDs recovered upon the introduction of unlabeled CP. The developed biosensor showed a limit of detection of 44 nM in a linear range of 0.10–0.54 μM and exhibited the strongest fluorescence intensity (about 47,000 a.u.) at 552 nm. This strategy was applied to detect purified CP in plant sap successfully with a recovery rate between 93–103%. Moreover, the feasibility of the proposed method was further verified by the detection of field samples, and the results were consistent with an enzyme-linked immunosorbent assay (ELISA). Contrarily to ELISA, the proposed biosensor did not require excessive washing and incubation steps, thus the detection could be rapidly accomplished in a few minutes. The high sensitivity and short assay time of this designed biosensor demonstrated its potential application in situ and rapid detection. In addition, the fluorescence quenching of CdTe QDs was attributed to dynamic quenching according to the Stern-Volmer equation.

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Section: Methodsmentioning

confidence: 99%

A Rapid, Fluorescence Switch-On Biosensor for Early Diagnosis of Sorghum Mosaic Virus

et al. 2022

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“…Hema et al [ 138 ] reported that IC-RT-PCR was 5,000 times more sensitive than DBIA and 10 times more sensitive than DB-PCR to detect SCSMV. Chen et al [ 139 ] demonstrated that the sensitivity of detecting SrMV by IC-RT-PCR 100-times higher than that of indirect ELISA and 1000-times higher than that of Dot-ELISA and therefore recommended IC-RT-PCR was more suitable for large volumes of samples. Subba et al [ 140 ] developed a D-IC-RT-PCR method that combined both serological and molecular methods to simultaneously detect and distinguish SCMV and SCSMV and was more sensitive than DAC-ELISA.…”

Section: Diagnosis/identificationmentioning

confidence: 99%

Sugarcane Mosaic Disease: Characteristics, Identification and Control

Wang

et al. 2021

Microorganisms

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Mosaic is one of the most important sugarcane diseases, caused by single or compound infection of Sugarcane mosaic virus (SCMV), Sorghum mosaic virus (SrMV), and/or Sugarcane streak mosaic virus (SCSMV). The compound infection of mosaic has become increasingly serious in the last few years. The disease directly affects the photosynthesis and growth of sugarcane, leading to a significant decrease in cane yield and sucrose content, and thus serious economic losses. This review covers four aspects of sugarcane mosaic disease management: first, the current situation of sugarcane mosaic disease and its epidemic characteristics; second, the pathogenicity and genetic diversity of the three viruses; third, the identification methods of mosaic and its pathogen species; and fourth, the prevention and control measures for sugarcane mosaic disease and potential future research focus. The review is expected to provide scientific literature and guidance for the effective prevention and control of mosaic through resistance breeding in sugarcane.

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“…Traditional detection methods used for SrMV include real-time fluorescent quantitative RT-PCR, conventional RT-PCR, reverse transcription loop-mediated amplification (RT-LAMP), and indirect ELISA detection [ 12 , 13 ]. However, these methods have limitations too, and the detection cycle might take up to an hour since they require sophisticated laboratory equipment and costly temperature-controlled amplification devices.…”

Section: Introductionmentioning

confidence: 99%

Optimization and Validation of Reverse Transcription Recombinase-Aided Amplification (RT-RAA) for Sorghum Mosaic Virus Detection in Sugarcane

Wang,

Liang,

et al. 2023

Pathogens

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Sorghum mosaic virus (SrMV) causes sugarcane mosaic disease and has significant adverse economic impacts on the cultivation of sugarcane. This study aimed to develop a rapid isotherm nucleic acid amplification method for detecting SrMV. Specific primers were designed to target the conserved region of the P3 gene of SrMV. The reverse transcription recombinase-aided amplification (RT-RAA) method was developed by screening primers and optimizing reaction conditions. Comparative analyses with RT-PCR demonstrated that the RT-RAA method exhibited superior specificity, sensitivity, and reliability for SrMV detection. Notably, using a standard plasmid diluted 10-fold continuously as a template, the sensitivity of RT-RAA was 100-fold higher than that of RT-PCR. Moreover, the RT-RAA reaction displayed flexibility in a temperature range of 24–49 °C, eliminating the need for expensive and complex temperature control equipment. Thus, this method could be utilized at ambient or even human body temperature. Within a short duration of 10 min at 39 °C, the target sequence of SrMV could be effectively amplified. Specificity analysis revealed no cross-reactivity between SrMV and other common sugarcane viruses detected via the RT-RAA. With its high sensitivity, rapid reaction time, and minimal equipment requirements, this method presents a promising diagnostic tool for the reliable and expedited detection of SrMV. Furthermore, it indicates broad applicability for successfully detecting other sugarcane viruses.

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Comparison of IC-RT-PCR, Dot-ELISA and Indirect-ELISA for the Detection of Sorghum Mosaic Virus in Field-Grown Sugarcane Plants

Cited by 4 publications

References 24 publications

A Rapid, Fluorescence Switch-On Biosensor for Early Diagnosis of Sorghum Mosaic Virus

A Rapid, Fluorescence Switch-On Biosensor for Early Diagnosis of Sorghum Mosaic Virus

Sugarcane Mosaic Disease: Characteristics, Identification and Control

Optimization and Validation of Reverse Transcription Recombinase-Aided Amplification (RT-RAA) for Sorghum Mosaic Virus Detection in Sugarcane

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