Quantitative in planta PCR assay for specific detection of Xanthomonas oryzae pv. oryzicola using putative membrane protein based primer set

2012 ◽  
Vol 40 ◽  
pp. 22-27 ◽  
Author(s):  
M.J. Kang ◽  
M.H. Kim ◽  
D.J. Hwang ◽  
M.S. Cho ◽  
Y. Seol ◽  
...  
Keyword(s):  
Membrane Protein ◽  
Xanthomonas Oryzae ◽  
In Planta ◽  
Specific Detection ◽  
Pcr Assay ◽  
Putative Membrane Protein

Development of a hemi-nested PCR assay for the specific detection ofMelissococcus pluton

1998 ◽  
Vol 37 (3) ◽  
pp. 165-174 ◽  
Author(s):  
Steven P Djordjevic ◽  
Kerrie Noone ◽  
Lisa Smith ◽  
Michael A Z Hornitzky
Keyword(s):  
Nested Pcr ◽  
Specific Detection ◽  
Pcr Assay

Identification and quantification of Fusarium oxysporum in planta and soil by means of an improved specific and quantitative PCR assay

2010 ◽  
Vol 46 (3) ◽  
pp. 372-382 ◽  
Author(s):  
Daniel Jiménez-Fernández ◽  
Miguel Montes-Borrego ◽  
Juan A. Navas-Cortés ◽  
Rafael M. Jiménez-Díaz ◽  
Blanca B. Landa
Keyword(s):  
Fusarium Oxysporum ◽  
Quantitative Pcr ◽  
In Planta ◽  
Pcr Assay ◽  
Quantitative Pcr Assay ◽  
Identification And Quantification

scholarly journals Pathogenicity and a TaqMan Real-Time PCR for Specific Detection of Pantoea allii, a Bacterial Pathogen of Onions

Plant Disease ◽  
2019 ◽  
Vol 103 (12) ◽  
pp. 3031-3040 ◽  
Author(s):  
Shabnam Rahimi-Khameneh ◽  
Sanni Hsieh ◽  
Renlin Xu ◽  
Tyler J. Avis ◽  
Sean Li ◽  
...  
Keyword(s):  
Real Time ◽  
Real Time Pcr ◽  
Phylogenetic Analyses ◽  
Similarity Index ◽  
Taqman Probe ◽  
Economic Losses ◽  
In Planta ◽  
Pcr Assay ◽  
Blind Test ◽  
Reliable Detection

Bacterial diseases of onion are reported to cause significant economic losses. Pantoea allii Brady, one of the pathogens causing the center rot on onions, has not yet been reported in Canada. We report the pathogenicity of P. allii on commercially available Canadian green onions (scallions). All P. allii-inoculated plants, irrespective of the inoculum concentration, exhibited typical leaf chlorotic discoloration on green onion leaves, which can reduce their marketability. Reisolation of P. allii from infected scallion tissues and reidentification by sequencing and phylogenetic analyses of the leuS gene suggest that the pathogen can survive in infected tissues 21 days after inoculation. This is the first report of P. allii as a potential pathogen of green onions. This study also reports the development and validation of a TaqMan real-time PCR assay targeting the leuS gene for reliable detection of P. allii in pure cultures and in planta. A 642-bp leuS gene fragment was targeted because it showed high nucleotide diversity and positively correlated with genome-based average nucleotide identity with respect to percent similarity index and identity of Pantoea species. The assay specificity was validated using 61 bacterial and fungal strains. Under optimal conditions, the selected primers and FAM-labeled TaqMan probe were specific for the detection of nine reference P. allii strains by real-time PCR. The 52 strains of other Pantoea spp. (n = 25), non-Pantoea spp. (n = 20), and fungi/oomycetes (n = 7) tested negative (no detectable fluorescence). Onion tissues spiked with P. allii, naturally infested onion bulbs, greenhouse infected green onion leaf samples, as well as an interlaboratory blind test were used to validate the assay specificity. The sensitivities of a 1-pg DNA concentration and 30 CFU are comparable to previously reported real-time PCR assays of other bacterial pathogens. The TaqMan real-time PCR assay developed in this study will facilitate reliable detection of P. allii and could be a useful tool for screening onion imports or exports for the presence of this pathogen.

scholarly journals A TaqMan-based real-time PCR assay for specific detection of novel duck reovirus in China

2020 ◽  
Vol 16 (1) ◽  
Author(s):  
Shuai Zhang ◽  
Weihua Li ◽  
Xiaodong Liu ◽  
Xudong Li ◽  
Bin Gao ◽  
...  
Keyword(s):  
Real Time ◽  
Real Time Pcr ◽  
Specific Detection ◽  
Pcr Assay

Specific detection of carcinoembryonic antigen-expressing tumor cells in bone marrow aspirates by polymerase chain reaction.

1994 ◽  
Vol 12 (4) ◽  
pp. 725-729 ◽  
Author(s):  
M Gerhard ◽  
H Juhl ◽  
H Kalthoff ◽  
H W Schreiber ◽  
C Wagener ◽  
...  
Keyword(s):  
Breast Cancer ◽  
Bone Marrow ◽  
Polymerase Chain Reaction ◽  
Tumor Cells ◽  
Normal Bone Marrow ◽  
Specific Detection ◽  
Chain Reaction ◽  
Pcr Assay ◽  
Normal Bone ◽  
Polymerase Chain

PURPOSE To establish a sensitive assay for the specific detection of carcinoembryonic antigen (CEA)-expressing tumor cells in the bone marrow of patients with colorectal cancer and other CEA-positive carcinomas. PATIENTS AND METHODS A CEA-specific nested reverse transcriptase (RT) polymerase chain reaction (PCR) assay was developed and optimized using limiting dilutions of a CEA-positive cancer cell line mixed with normal bone marrow cell specimens. The optimized test was then used to examine bone marrow samples obtained from 15 patients with abdominal carcinomas (colorectal, n = 10; pancreatic, n = 3; gastric, n = 2) and six patients with breast cancer. Specificity was assessed by examination of 56 negative controls (malignant hematologic disease, n = 28; nonmalignant disease conditions, n = 5; healthy bone marrow donors, n = 8; normal peripheral-blood samples, n = 15). For 11 patients with abdominal carcinomas, immunostaining evaluations were performed using an anti-CEA and an anticytokeratin antibody, and the results compared with the nested PCR assay. RESULTS In the sensitivity calibration system, single CEA-expressing tumor cells were detected in 2 to 5 x 10(7) normal bone marrow cells. All 56 control samples failed to amplify. This demonstrates that mRNAs coding for highly homologous CEA-related antigens expressed by various lineages of blood cells do not interfere. Bone marrow samples from 10 of 15 patients with abdominal cancers and four of six breast cancer patients scored positive, indicating micrometastatic bone disease. Four of 11 samples from the gastrointestinal cancer patients were found to be positive by the PCR method, but were negative with the immunocytology method. CONCLUSION Since approximately 30% of the colorectal carcinoma patients that score negative in immunocytology staining of bone marrow samples have been reported to relapse, earlier diagnosis of the presence of malignant cells is needed. Our result that samples scoring positive in the described CEA-specific PCR test remained negative by two immunostaining methods suggests a higher sensitivity. We conclude that PCR amplification of CEA mRNA may lead to an earlier diagnosis of micrometastatic bone disease in patients with CEA-expressing carcinomas.

Development of a novel dual priming oligonucleotide system‐based PCR assay for specific detection of Salmonella from food samples

2020 ◽  
Vol 40 (3) ◽  
Author(s):  
Dan‐Dan Li ◽  
Chun‐Bo Hao ◽  
Zhong‐Mei Liu ◽  
Sui‐Jia Wang ◽  
Yu Wang ◽  
...  
Keyword(s):  
Food Samples ◽  
Specific Detection ◽  
Pcr Assay ◽  
Dual Priming Oligonucleotide

scholarly journals Gene Involved in Transcriptional Activation of the hrp Regulatory Gene hrpG in Xanthomonas oryzae pv. oryzae

2006 ◽  
Vol 188 (11) ◽  
pp. 4158-4162 ◽  
Author(s):  
Seiji Tsuge ◽  
Takeshi Nakayama ◽  
Shinsaku Terashima ◽  
Hirokazu Ochiai ◽  
Ayako Furutani ◽  
...  
Keyword(s):  
Gene Expression ◽  
Plant Pathogen ◽  
Transcriptional Activation ◽  
Regulatory Gene ◽  
Xanthomonas Oryzae ◽  
In Planta

ABSTRACT A novel regulatory gene, trh, which is involved in hrp gene expression, is identified in the plant pathogen Xanthomonas oryzae pv. oryzae. In the trh mutant, expression of HrpG, which is a key regulator for hrp gene expression, is reduced both under the in vitro hrp-inducing condition and in planta.

Characterization of a Gene Coamplified with Ki-ras in Y1 Murine Adrenal Carcinoma Cells That Codes for a Putative Membrane Protein

Genomics ◽  
1994 ◽  
Vol 20 (2) ◽  
pp. 227-230 ◽  
Author(s):  
Alan F. Scott ◽  
Andrew Elizaga ◽  
James Morrell ◽  
Andrew Bergen ◽  
Margaret B. Penno
Keyword(s):  
Membrane Protein ◽  
Carcinoma Cells ◽  
Adrenal Carcinoma ◽  
Putative Membrane Protein
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