A Quantative Real-Time PCR of Phytophthora infestans in different Indian potato cultivars

Hussain, Touseef; Anwar, Firoz

doi:10.9790/2380-0441726

Cited by 3 publications

(2 citation statements)

References 43 publications

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“…Previously, several quantitative assays have been reported for P. infestans, based on either TaqMan probe or SYBR Green dye. , However, TaqMan probe-based assays are very expensive, and SYBR Green dye suffers several disadvantages compared to EvaGreen . For instance, EvaGreen dye shows lower background fluorescence and less inhibitory effect in the PCR reaction compared to that of SYBR Green .…”

Section: Resultsmentioning

confidence: 99%

Extraction of Plant DNA by Microneedle Patch for Rapid Detection of Plant Diseases

Paul¹,

Saville²,

Hansel³

et al. 2019

ACS Nano

135

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In-field molecular diagnosis of plant diseases via nucleic acid amplification is currently limited by cumbersome protocols for extracting and isolating pathogenic DNA from plant tissues. To address this challenge, a rapid plant DNA extraction method was developed using a disposable polymeric microneedle (MN) patch. By applying MN patches on plant leaves, amplification-assay-ready DNA can be extracted within a minute from different plant species. MNextracted DNA was used for direct polymerase chain reaction amplification of plant plastid DNA without purification. Furthermore, using this patch device, extraction of plant pathogen DNA (Phytophthora infestans) from both laboratoryinoculated and field-infected leaf samples was performed for detection of late blight disease in tomato. MN extraction achieved 100% detection rate of late blight infections for samples after 3 days of inoculation when compared to the conventional gold standard cetyltrimethylammonium bromide (CTAB)-based DNA extraction method and 100% detection rate for all blind field samples tested. This simple, cell-lysis-free, and purification-free DNA extraction method could be a transformative approach to facilitate rapid sample preparation for molecular diagnosis of various plant diseases directly in the field.

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Section: Resultsmentioning

confidence: 99%

Extraction of Plant DNA by Microneedle Patch for Rapid Detection of Plant Diseases

Paul¹,

Saville²,

Hansel³

et al. 2019

ACS Nano

135

View full text Add to dashboard Cite

show abstract

“…Quantative Real-Time PCR is a new advance in PCR methodology. Several studies have demonstrated that the sensitivity of Real-Time PCRcan also exceed that of conventional PCR by up to 1000 times (Hu et al 1993, Mumford et al 2004, Ratti et al 2004, Hussain et al 2013b, 2014. In addition, Real-time PCR can also be used to quantify target nucleic acid in a range of sample types.…”

Section: Open Accessmentioning

confidence: 99%

Soil Baiting, Rapid PCR Assay and Quantitative Real Time PCR to Diagnose Late Blight of Potato in Quarantine Programs

Hussain

Singh

Anwar

2018

J. Nep. Agric. Res. Counc.

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Phytophthora infestans (mont) de Bary is a pathogen of great concern across the globe, and accurate detection is an important component in responding to the outbreaks of potential disease. Although the molecular diagnostic protocol used in regulatory programs has been evaluated but till date methods implying direct comparison has rarely used. In this study, a known area soil samples from potato fields where light blight appear every year (both A1 and A2 mating type) was assayed by soil bait method, PCR assay detection and quantification of the inoculums. Suspected disease symptoms appeared on bait tubers were further confirmed by rapid PCR, inoculums were quantified through Real Time PCR, which confirms presence of P. infestans. These diagnostic methods can be highly correlated with one another. Potato tuber baiting increased the sensitivity of the assay compared with direct extraction of DNA from tuber and soil samples. Our study determines diagnostic sensitivity and specificity of the assays to determine the performance of each method. Overall, molecular techniques based on different types of PCR amplification and Real-time PCR can lead to high throughput, faster and more accurate detection method which can be used in quarantine programmes in potato industry and diagnostic laboratory.

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Molecular Diagnosis of Killer Pathogen of Potato: Phytophthora infestans and Its Management

Hussain

Singh

2016

Fungal Biology

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A Quantative Real-Time PCR of Phytophthora infestans in different Indian potato cultivars

Cited by 3 publications

References 43 publications

Extraction of Plant DNA by Microneedle Patch for Rapid Detection of Plant Diseases

Extraction of Plant DNA by Microneedle Patch for Rapid Detection of Plant Diseases

Soil Baiting, Rapid PCR Assay and Quantitative Real Time PCR to Diagnose Late Blight of Potato in Quarantine Programs

Molecular Diagnosis of Killer Pathogen of Potato: Phytophthora infestans and Its Management

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