scholarly journals Applying Real-Time Quantitative PCR to Fusarium Crown Rot of Wheat

Plant Disease ◽  
2007 ◽  
Vol 91 (8) ◽  
pp. 1021-1028 ◽  
Author(s):  
A. C. Hogg ◽  
R. H. Johnston ◽  
A. T. Dyer
Keyword(s):  
Real Time ◽  
Quantitative Pcr ◽  
Field Experiments ◽  
Qpcr Assay ◽  
Crown Rot ◽  
Yield Reduction ◽  
Fusarium Crown Rot ◽  
Persistent Problem ◽  
Real Time Quantitative Pcr ◽  
Severity Scores

Fusarium crown rot (FCR) of wheat is a persistent problem that causes significant losses worldwide. In Montana, FCR is caused primarily by Fusarium culmorum and F. pseudograminearum. Recently, a real-time quantitative PCR (QPCR) assay was developed for FCR using primers and probes specific for a segment of the trichodiene synthase (tri5) gene. The purpose of this study was to determine the utility of QPCR for accessing FCR severity on wheat in field experiments. In 2004 and 2005, plots of spring and durum wheat were inoculated with varying levels of F. pseudograminearum oat inoculum and grown under rain-fed conditions. Two weeks prior to harvest, plants were collected from the plots and assessed for FCR severity and analyzed by QPCR for Fusarium DNA quantities. Disease severity scores (DSS) and Fusarium DNA quantities were positively correlated with each other for all three cultivars in 2004 but for only the durum cultivar in 2005 (P < 0.05). In 2004, grain yields for both spring wheat cultivars were negatively correlated with Fusarium DNA quantities (P > 0.05). When DSS and Fusarium DNA quantities negatively correlated with yield, both measurements were comparable in predicting yield reduction (R = –0.64 and –0.77, respectively). Results indicate that this QPCR assay is effective in measuring FCR severity in wheat.

scholarly journals Accurate, Fast and Cost-Effective Diagnostic Test for Monosomy 1p36 Using Real-Time Quantitative PCR

2014 ◽  
Vol 2014 ◽  
pp. 1-8
Author(s):  
Pricila da Silva Cunha ◽  
Heloisa B. Pena ◽  
Carla Sustek D’Angelo ◽  
Celia P. Koiffmann ◽  
Jill A. Rosenfeld ◽  
...  
Keyword(s):  
Real Time ◽  
Diagnostic Test ◽  
Quantitative Pcr ◽  
Target Genes ◽  
Cost Effective ◽  
Qpcr Assay ◽  
Deletion Syndrome ◽  
Comparative Genomic ◽  
Real Time Quantitative Pcr ◽  
Subtelomeric Deletion

Monosomy 1p36 is considered the most common subtelomeric deletion syndrome in humans and it accounts for 0.5–0.7% of all the cases of idiopathic intellectual disability. The molecular diagnosis is often made by microarray-based comparative genomic hybridization (aCGH), which has the drawback of being a high-cost technique. However, patients with classic monosomy 1p36 share some typical clinical characteristics that, together with its common prevalence, justify the development of a less expensive, targeted diagnostic method. In this study, we developed a simple, rapid, and inexpensive real-time quantitative PCR (qPCR) assay for targeted diagnosis of monosomy 1p36, easily accessible for low-budget laboratories in developing countries. For this, we have chosen two target genes which are deleted in the majority of patients with monosomy 1p36:PRKCZandSKI. In total, 39 patients previously diagnosed with monosomy 1p36 by aCGH, fluorescentin situhybridization (FISH), and/or multiplex ligation-dependent probe amplification (MLPA) all tested positive on our qPCR assay. By simultaneously using these two genes we have been able to detect 1p36 deletions with 100% sensitivity and 100% specificity. We conclude that qPCR ofPRKCZandSKIis a fast and accurate diagnostic test for monosomy 1p36, costing less than 10 US dollars in reagent costs.

scholarly journals Development of Conventional and Real-Time Quantitative PCR Assays for Diagnosis and Monitoring of Scabies

2015 ◽  
Vol 53 (7) ◽  
pp. 2095-2102 ◽  
Author(s):  
Samson S. Y. Wong ◽  
Rosana W. S. Poon ◽  
Sandy Chau ◽  
Sally C. Y. Wong ◽  
Kelvin K. W. To ◽  
...  
Keyword(s):  
Real Time ◽  
Quantitative Pcr ◽  
Copy Number ◽  
Mitochondrial Gene ◽  
Qpcr Assay ◽  
Dna Copy Number ◽  
Predictive Values ◽  
Content Type ◽  
Lesional Skin ◽  
Real Time Quantitative Pcr

Scabies remains the most prevalent, endemic, and neglected ectoparasitic infestation globally and can cause institutional outbreaks. The sensitivity of routine microscopy for demonstration ofSarcoptes scabieimites or eggs in skin scrapings is only about 50%. Except for three studies using conventional or two-tube nested PCR on a small number of cases, no systematic study has been performed to improve the laboratory diagnosis of this important infection. We developed a conventional and a real-time quantitative PCR (qPCR) assay based on the mitochondrial cytochromecoxidase subunit 1 (cox1) gene ofS. scabiei. Thecox1gene is relatively well conserved, with its sequence having no high levels of similarity to the sequences of other human skin mites, pathogenic zoonotic mites, or common house dust mite species. This mitochondrial gene is also present in large quantities in arthropod cells, potentially improving the sensitivity of a PCR-based assay. In our study, both assays were specific and were more sensitive than microscopy in diagnosing scabies, with positive and negative predictive values of 100%. TheS. scabieiDNA copy number in the microscopy-positive specimens was significantly higher than that in the microscopy-negative specimens (medianS. scabieiDNA copy number, 3.604 versus 2.457 log10copies per reaction;P= 0.0213). In the patient with crusted scabies, the qPCR assay performed on lesional skin swabs instead of scrapings revealed that the parasite DNA load took about 2 weeks to become negative after treatment. The utility of using lesional skin swabs as an alternative sample for diagnosis of scabies by PCR should be further evaluated.

scholarly journals Quantification and Stability of Human Adenoviruses and Polyomavirus JCPyV in Wastewater Matrices

2006 ◽  
Vol 72 (12) ◽  
pp. 7894-7896 ◽  
Author(s):  
Silvia Bofill-Mas ◽  
Nestor Albinana-Gimenez ◽  
Pilar Clemente-Casares ◽  
Ayalkibet Hundesa ◽  
Jesus Rodriguez-Manzano ◽  
...  
Keyword(s):  
Real Time ◽  
Quantitative Pcr ◽  
High Stability ◽  
Wastewater Sludge ◽  
Human Polyomavirus ◽  
Sewage Effluent ◽  
Viral Contamination ◽  
Human Adenoviruses ◽  
Urban Sewage ◽  
Real Time Quantitative Pcr

ABSTRACT Human adenoviruses (HAdV) and human polyomavirus JCPyV have been previously proposed as indicators of fecal viral contamination in the environment. Different wastewater matrices have been analyzed by applying real-time quantitative PCR procedures for the presence, quantity, and stability of a wide diversity of excreted HAdV and JCPyV. High quantities of HAdV and JCPyV were detected in sewage, effluent wastewater, sludge, and biosolid samples. Both viruses showed high stability in urban sewage. These results confirm the suitability of both viruses as indicators of human fecal viral pollution.

Analysis of Caecal Microbiota in Rats Fed with Genetically Modified Rice by Real-Time Quantitative PCR

2011 ◽  
Vol 76 (1) ◽  
pp. M88-M93 ◽  
Author(s):  
Wentao Xu ◽  
Liting Li ◽  
Jiao Lu ◽  
YunBo Luo ◽  
Ying Shang ◽  
...  
Keyword(s):  
Real Time ◽  
Quantitative Pcr ◽  
Genetically Modified ◽  
Genetically Modified Rice ◽  
Real Time Quantitative Pcr ◽  
Caecal Microbiota
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