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Quantitative Detection (quantitative + detection)
Selected AbstractsQuantitative Detection of Bioassays with a Low-Cost Image-Sensor Array for Integrated Microsystems,ANGEWANDTE CHEMIE, Issue 41 2009Daynene Tragbare Assaysysteme erfordern kostengünstige Detektoren zur Kopplung mit einer mikrofluidikbasierten Probenbehandlung. Ein gewöhnlicher Digitalkamerasensor wurde nun mit verschiedenen Lab-on-a-Chip-Architekturen verschaltet und zur Abbildung von nanolitergroßen Reagenströpfchen sowie zur quantitativen Photometrie von kolorimetrischen und Biolumineszenz-Assays eingesetzt (siehe Kontaktbilder der mikrofluidischen Elektrodenanordnung (rechts) und der kolorimetrischen (oben) und biolumineszenten Proben (unten)). [source] Quantitative detection of fluid distribution using time-lapse seismicGEOPHYSICAL PROSPECTING, Issue 2 2007Futoshi Tsuneyama ABSTRACT Although previous seismic monitoring studies have revealed several relationships between seismic responses and changes in reservoir rock properties, the quantitative evaluation of time-lapse seismic data remains a challenge. In most cases of time-lapse seismic analysis, fluid and/or pressure changes are detected qualitatively by changes in amplitude strength, traveltime and/or Poisson's ratio. We present the steps for time-lapse seismic analysis, considering the pressure effect and the saturation scale of fluids. We then demonstrate a deterministic workflow for computing the fluid saturation in a reservoir in order to evaluate time-lapse seismic data. In this approach, we derive the physical properties of the water-saturated sandstone reservoir, based on the following inputs: VP, VS, , and the shale volume from seismic analysis, the average properties of sand grains, and formation-water properties. Next, by comparing the in-situ fluid-saturated properties with the 100% formation-water-saturated reservoir properties, we determine the bulk modulus and density of the in-situ fluid. Solving three simultaneous equations (relating the saturations of water, oil and gas in terms of the bulk modulus, density and the total saturation), we compute the saturation of each fluid. We use a real time-lapse seismic data set from an oilfield in the North Sea for a case study. [source] Quantitative detection of hepatitis B virus DNA in serum by a new rapid real-time fluorescence PCR assayJOURNAL OF VIRAL HEPATITIS, Issue 6 2001R. Jardi A sensitive and accurate HBV DNA quantification assay is essential for monitoring hepatitis B virus (HBV) replication. This study evaluated a real-time PCR method performed in the LightCyclerTM analyser for quantitative HBV DNA assay. HBV DNA results with this method were compared with those obtained using a branched-chain DNA (bDNA) solution hybridization assay. Real-time PCR was performed using two adjacent fluorescently labelled probes and primers corresponding to the HBV core gene. The same standard employed in the bDNA assay was used for calibration. Serum samples came from 193 HBV surface antigen (HBsAg)-positive patients (34 HBV e antigen (HBeAg)-positive and 93 with antibody to HBeAg (anti-HBe)), and 66 asymptomatic HBV carriers. In addition, we analysed serum samples from 8 anti-HBe-positive patients who had been receiving lamivudine treatment for more than three years. A linear standard curve was seen in the range from 103 to 108 copies/mL. In the reproducibility analysis, intra-assay coefficient of variation (CVs) at two known HBV DNA concentrations were 4% and 2% and interassay CVs were 6% and 4%. The median of serum HBV DNA by real-time PCR was 9.2 × 108 copies/mL in HBeAg-positive patients with persistently elevated alanine aminotransferase (ALT) levels, 1.3 × 107 copies/mL in anti-HBe-positive cases with persistently elevated ALT levels, 3.7 × 104 copies/mL in anti-HBe-positive patients with fluctuating ALT levels and 104 copies/mL in asymptomatic HBV carriers. The differences in HBV DNA levels among the various groups studied were statistically significant (P < 0.05). The cut-off between chronic hepatitis patients and asymptomatic carriers was found to be at a serum HBV DNA concentration of 5 × 104 copies/mL. Of the 109 serum samples with a viral load < 7.5 × 105 (negative by bDNA assay) 44 (40%) were positive by real-time PCR: 24 (56%) chronic hepatitis and 20 (33%) asymptomatic carriers. There was a positive association between HBV DNA levels determined by real-time PCR and ALT levels (P < 0.05), which was not observed with the bDNA assay for HBV DNA quantification. At 12 months of lamivudine treatment, 6 patients (75%) showed HBV DNA levels < 5 × 104 copies/mL (range < 103,2 × 103), significantly lower than at baseline. At 36 months, 2 of 8 (25%) showed HBV DNA levels persistently lower than 5 × 104 copies/mL (1.7 × 103, 6 × 103). The LightCycler quantitative real-time PCR is a practical, sensitive, reproducible single-tube assay with a wide dynamic range of detection. The assay is automatic except for DNA extraction and the running time is only 70 min. The LightCycler real-time PCR is useful for identifying different states of HBV infection and for evaluating the efficacy of viral therapy. [source] Expression of insulin-like growth factor-I in lesional and non-lesional skin of patients with morphoeaBRITISH JOURNAL OF DERMATOLOGY, Issue 1 2008M.M.T. Fawzi Summary Background, Morphoea (scleroderma) is a chronic disorder characterized by circumscribed sclerotic plaques with the hallmark of increased fibroblast activation and fibrosis. Through its effect on connective tissue cells and immune cells, insulin-like growth factor (IGF)-I has been found to play a role in some autoimmune connective tissue diseases and has been implicated in the pathogenesis of several fibrotic disorders. Objectives, To evaluate the role of IGF-I in the pathogenesis of morphoea. Methods, The study was carried out on 15 patients with morphoea and nine healthy controls. Two 5-mm punch skin biopsies were taken from every patient (one from lesional and one from non-lesional skin) and a single biopsy was taken from the normal skin of each control. A 10-mL blood sample was also taken from each patient and control. Quantitative detection of tissue and serum levels of IGF-I was done using an enzyme-linked immunosorbent assay technique. Results, IGF-I in lesional skin was significantly higher than in non-lesional and control skin (P = 0·001 and P = 0·021, respectively). Moreover, a significantly higher level of IGF-I was detected in patient serum when compared with control serum (P < 0·001). A direct significant correlation existed between lesional and non-lesional skin level (r = 0·618, P = 0·014), and between lesional skin level and Rodnan score (r = 0·538, P = 0·039). Conclusions, Despite the small sample size, this study suggests that IGF-I plays an important role in the pathogenesis of fibrosis, characteristic of morphoea. Studies on a larger number of patients with morphoea as well as on patients with systemic sclerosis are recommended. Furthermore, therapeutic trials using IGF-I antagonist (octreotide) are highly recommended in patients with morphoea. [source] Application of Exchangeable Biochemical Reactors with Oxidase-Catalase-Co-immobilizates and Immobilized Microorganisms in a Microfluidic Chip-CalorimeterENGINEERING IN LIFE SCIENCES (ELECTRONIC), Issue 5 2008M. Leifheit Abstract Several methods for the quantitative detection of different compounds, e.g., L -amino acids, sugars or alcohols in liquid media were developed by application of an automatic measuring unit including a fluid chip-calorimeter FCC-21. For this purpose, enzymes were immobilized covalently on the inner and outer surface of CPG (controlled porous glass)-spherules with an outer diameter of 100,,m and filled into a micro flow-through reaction chamber (VR = 20,,L). The design of the measuring cell allows for easy insertion into the calorimeter device of a stored series of comfortably pre-fabricated measuring cells. These cells can be filled with different enzyme immobilizates. Different oxidases were used and co-immobilized with catalase for the improvement of the detection sensitivity. A signal amplification could be achieved up to a factor of 3.5 with this configuration. ,- D -glucose, ethanol and L -lysine could be detected in a range of 0.25,1.75,mM using glucose oxidase, alcohol oxidase and lysine oxidase. The group of oxidases in combination with the enzymatic catalysis of the intermediate H2O2 allows the quantitative detection of a large number of analytes. A good measurement and storage stability could be achieved for several weeks by this immobilization method. In addition to enzyme-based detection reactions, it was shown that living microorganisms can be immobilized in the reaction chamber. Thus, the system can be used as a whole-cell biosensor. The quantitative detection of phenol in the range of 10,100,,M could be performed using the actinomycete Rhodococcus sp. immobilized on glass beads by means of embedding into polymers. [source] Laser-induced breakdown spectroscopy for soil diagnosticsEUROPEAN JOURNAL OF SOIL SCIENCE, Issue 2 2001J. Bublitz Summary Laser-optical measurements and fibre optics are potentially attractive tools for applications in soil science because of their great sensitivity and selectivity and their capabilities for on-line and in situ analysis. We have investigated laser-induced breakdown spectroscopy (LIBS) for the quantitative detection of metal ions on the surface of natural soil samples from two sites (Hohenschulen and Oderbruch, Germany). The LIBS technique allows the spatially resolved investigation of adsorption and desorption effects of ions in soil. A frequency doubled (532 nm) and Q-switched Nd:YAG laser with a pulse duration of 8 ns is focused on the soil surface and induces a plasma. Typical power densities are 150 mJ mm,2. The plasma emission is recorded in time and spectrally resolved by a gateable optical multichannel analyser (OMA). A delay time of about 500 ns between laser pulse and OMA gate was used to resolve single atomic and ionic spectral lines from the intense and spectrally broad light that is emitted by the plasma itself. The dependency of the LIBS signal of a single spectral line on the amount of water in the sample is investigated in detail. The results indicate that quenching of water in the plasma plume reduces the line intensities, while the interaction with aquatic colloids increases the intensity. The two processes compete with each other, and a non-linear correlation between measured line intensities and the amount of water in the sample is obtained. This is verified by a simple computer simulation and has to be taken into account for the quantitative interpretation of LIBS signals, e.g. when absolute concentrations are estimated. In the present investigation natural calcium concentrations <,2 ,g kg,1 were measured with the LIBS technique in the samples for the two test sites. In addition, measurements were made with dry and water-saturated BaCl2 mixed soil samples, and no significant difference in the detection limit for barium was obtained. [source] Application of real-time PCR for quantitative detection of Campylobacter jejuni in poultry, milk and environmental waterFEMS IMMUNOLOGY & MEDICAL MICROBIOLOGY, Issue 3 2003Chengbo Yang Abstract Campylobacter jejuni is a leading human food-borne pathogen. The rapid and sensitive detection of C. jejuni is necessary for the maintenance of a safe food/water supply. In this article, we present a real-time polymerase chain reaction (PCR) assay for quantitative detection of C. jejuni in naturally contaminated poultry, milk and environmental samples without an enrichment step. The whole assay can be completed in 60 min with a detection limit of approximately 1 CFU. The standard curve correlation coefficient for the threshold cycle versus the copy number of initial C. jejuni cells was 0.988. To test the PCR system, a set of 300 frozen chicken meat samples, 300 milk samples and 300 water samples were screened for the presence of C. jejuni. 30.6% (92/300) of chicken meat samples, 27.3% (82/300) of milk samples, and 13.6% (41/300) of water samples tested positive for C. jejuni. This result indicated that the real-time PCR assay provides a specific, sensitive and rapid method for quantitative detection of C. jejuni. Moreover, it is concluded that retail chicken meat, raw milk and environmental water are commonly contaminated with C. jejuni and could serve as a potential risk for consumers in eastern China, especially if proper hygienic and cooking conditions are not maintained. [source] A Direct, Multiplex Biosensor Platform for Pathogen Detection Based on Cross-linked Polydiacetylene (PDA) SupramoleculesADVANCED FUNCTIONAL MATERIALS, Issue 23 2009Cheol Hee Park Abstract This study focuses on the development of a multiplex pathogen-detection platform based on polydiacetylene (PDA) using a novel immobilization procedure. PDA liposome-based solid sensors have a critical drawback as the PDA liposomes are not stably immobilized onto the solid substrate. Therefore, to overcome this problem, an interlinker, ethylenediamine, is introduced, which acts as a cross-linker between individual PDA liposomes. The quantity of ethylenediamine added was optimized to 1,mM, as measured by the fluorescence signal emitted by the stably immobilized PDA liposomes, a concentration at which the fluorescence signal is 10 times higher than for the resulting PDA chips made without the interlinker. This procedure is used to manufacture PDA liposome-based multiplex biosensor arrays for well-known water and food-borne pathogens. The fabricated biosensor was able to perform the simultaneous and quantitative detection of 6 species of pathogens. As such, the results demonstrated from this research can be exploited for the development of more advanced PDA-based biosensors and diagnostics. [source] smcL as a novel diagnostic marker for quantitative detection of Listeria ivanovii in biological samplesJOURNAL OF APPLIED MICROBIOLOGY, Issue 3 2010D. Rodríguez-Lázaro Abstract Aims:, To develop a novel molecular tool for the quantitative detection of the ruminant pathogen Listeria ivanovii in different biological matrices. Methods and Results:, A real-time PCR (RTi-PCR) for the quantitative and species-specific identification of L. ivanovii was designed to target the region of the smcL gene. The assay includes an internal amplification control (IAC) to avoid false-negative results. The smcL -IAC RTi-PCR assay was 100% selective and allowed the detection of as little as one genome equivalent in 45% of reactions. The quantification accuracy was excellent, as demonstrated by its high linearity (R2 > 0·9989) and PCR efficiency (E > 0·984) over a 6-log dynamic range, down to 10 genome equivalents. Finally, the applicability of this assay was evaluated with artificially contaminated biological matrices implicated in the transmission of this bacterium such as sheep raw milk, blood and amniotic fluid. The smcL -IAC RTi-PCR assay allowed the detection of as few as 50 colony forming unit numbers (CFUs) per 25 ml of raw milk, 43 CFUs per 1 ml of blood or 50 CFUs per 1 ml of amniotic fluid. Conclusions:, This method can be an adequate alternative for the identification of L. ivanovii and for complete diagnosis of animal and human listeriosis. Significance and Impact of the Study:, We present an alternative for the detection of another pathogenic member of Listeria genus, which can help to distinguish from Listeria monocytogenes and therefore facilitates the establishment of preventive and prophylactic measures in food and farm environments. [source] Development and comparison of SYBR Green quantitative real-time PCR assays for detection and enumeration of sulfate-reducing bacteria in stored swine manureJOURNAL OF APPLIED MICROBIOLOGY, Issue 6 2008C. Spence Abstract Aims:, To develop and evaluate primer sets targeted to the dissimilatory sulfite reductase gene (dsrA) for use in quantitative real-time PCR detection of sulfate-reducing bacteria (SRB) in stored swine manure. Methods and Results:, Degenerate primer sets were developed to detect SRB in stored swine manure. These were compared with a previously reported primer set, DSR1F+ and DSR-R, for their coverage and ability to detect SRB communities in stored swine manure. Sequenced clones were most similar to Desulfovibrio sp. and Desulfobulbus sp., and these SRB populations differed within different manure ecosystems. Sulfur content of swine diets was shown to affect the population of Desulfobulbus -like Group 1 SRB in manure. Conclusions:, The newly developed assays were able to enumerate and discern different groups of SRB, and suggest a richly diverse and as yet undescribed population of SRB in swine manure. Significance and Impact of the Study:, The PCR assays described here provide improved and efficient molecular tools for quantitative detection of SRB populations. This is the first study to show population shifts of SRB in swine manure, which are a result of either the effects of swine diets or the maturity of the manure ecosystem. [source] A colony immunoblotting method for quantitative detection of a Bifidobacterium animalis probiotic strain in human faecesJOURNAL OF APPLIED MICROBIOLOGY, Issue 6 2000H. Duez A colony immunoblotting method has been developed to allow detection of the probiotic Bifidobacterium animalis strain DN-173 010 in human faecal samples. Rabbits were immunized with heat-killed DN-173 010 bacteria resulting in the production of an antiserum highly specific for bacteria belonging to Bif. animalis species. Of the 89 strains representative of 29 different bifidobacterial species tested, only the 15 strains of the Bif. animalis species could be detected with the antiserum. In Western immunoblotting the serum reacts with a protein of 45-kDa apparent molecular weight. None of the bacteria classically encountered in human faecal samples and able to grow on non-selective Columbia blood agar (enterobacteria, Bacteroides or Lactobacillus for instance) reacted with the antiserum. Taking advantage of the high specificity of the antiserum and of the absence of Bif. animalis bacteria in faeces samples of five human volunteers, we demonstrated that strain DN-173 010 survives the intestinal transit. Being based on a combination of semiselective cultivation and colony immunoblotting techniques, the method allowed detection of the Bif. animalis strain even when it represented only one thousandth of the total bifidobacterial population. [source] Quantitative PCR determination of human cytomegalovirus in blood cellsJOURNAL OF CLINICAL LABORATORY ANALYSIS, Issue 3 2001J. Satou Abstract We evaluated a rapid and sensitive method to determine human cytomegalovirus (CMV) DNA levels in blood cells using a quantitative polymerase chain reaction (PCR) technique. This method is based on real-time detection of PCR using a dual fluorescence-labeled probe and a sequence detector. Ten copies of CMV DNA were detected, when 1 ,g of DNA from blood samples was used with this method, and a good correlation was obtained between increased concentrations of copy numbers calculated and measured copy numbers of CMV DNA (r = 0.999). Forty normal subjects exhibited no copies of CMV DNA. On the other hand, a 6-month-old girl tested positive for increased levels 4 weeks after liver transplant. This method is simple, accurate, and sensitive for the quantitative detection of CMV DNA in vivo, indicating possible applications for the diagnosis and monitoring of CMV infection. J. Clin. Lab. Anal. 15:122,126, 2001. © 2001 Wiley-Liss, Inc. [source] Real time PCR detection of Piscirickettsia salmonis from formalin-fixed paraffin-embedded tissuesJOURNAL OF FISH DISEASES, Issue 10 2008S Karatas Abstract Piscirickettsia salmonis is the causative agent of piscirickettsiosis, a transmissible disease of salmonid fish. Diagnosis of piscirickettsiosis has traditionally been based upon identification of typical pathological changes by histological investigation, with confirmation by immunohistochemistry on formalin-fixed, paraffin-embedded tissues. However, implementation of more rapid confirmatory techniques, preferably with higher levels of sensitivity and possibilities for quantification, is desirable. A real-time polymerase chain reaction (PCR) assay was designed for specific detection of P. salmonis and tested on samples extracted from formalin-fixed paraffin-embedded material. Construction of a PCR-target mimic allowed determination of detection limits, linearity of the real-time PCR and quantitative detection of P. salmonis. The present study demonstrates the capability of the described real time PCR assay for detection of P. salmonis from paraffin-embedded material with a high degree of sensitivity and specificity. Implementation of this assay constitutes an important development for a rapid and secure diagnosis of piscirickettsiosis. [source] APPLICATION OF STEPWISE AMMONIUM SULFATE PRECIPITATION AS CLEANUP TOOL FOR AN ENZYME-LINKED IMMUNOSORBENT ASSAY OF GLYPHOSATE OXIDOREDUCTASE IN GENETICALLY MODIFIED RAPE OF GT73JOURNAL OF FOOD BIOCHEMISTRY, Issue 5 2009WENTAO XU ABSTRACT The method of enzyme-linked immunosorbent assay after stepwise ammonium sulfate (AS) purification (AS-ELISA) was developed and used to detect genetically modified (GM) rape of GT73 containing glyphosate oxidoreductase (Gox). Gox protein encoded by the Gox gene from Achromobacter sp. was highly expressed as inclusion bodies in Escherichia coli BL21 (DE3) and purified to homogeneity by Ni2+affinity chromatography. A simple and efficient extraction and purification procedure of Gox protein from the seeds and leaves of GM rape was developed by means of stepwise AS precipitation. Purified polyclonal antibodies against Gox was produced and enzyme-linked immunosorbent assay (ELISA) procedures were established further on to measure the Gox protein. AS-ELISA allowed 5% GMOs to be detected in the seeds of GT73 and 0.5% GMOs to be detected in the leaves of GT73 rape, which makes this method an acceptable method to access Gox protein in GM rape of GT73. PRACTICAL APPLICATIONS Many GMOs containing Gox gene have been approved worldwide such as GT73 rape, 1,445 cotton and Mon832 maize. Protein based methods were more important than DNA based methods, because protein performs a specific and concrete function and is closely interconnected with crop traits. AS-ELISA method can be used in the screening of GM plant, Gox protein expression assay and quantitative detection for GMO labeling. AS-ELISA Gox detecting method was established in this paper and was being evaluated of Inter-laboratory Comparison in some of Chinese GMO detection and assessment centers. With the knowledge of ELISA, ELISA method will be the national standards and international and will be a beneficial supplement for the DNA based GMO detecting methods. [source] Analysis of intracellular short organic acid-coenzyme A esters from actinomycetes using liquid chromatography-electrospray ionization-mass spectrometryJOURNAL OF MASS SPECTROMETRY (INCORP BIOLOGICAL MASS SPECTROMETRY), Issue 9 2007Je Won Park Abstract A method employing silicone oil density centrifugation, solid-phase extraction (SPE) cleanup, and LC-ESI-MS/MS analysis was developed for the rapid, selective, sensitive, and quantitative detection of an intracellular pool of short organic acid-CoA esters in actinomycetes. The detection limit was determined to be approximately 0.8 pmol (1.2 ng/ml) for each standard CoA-ester analyzed by the present LC-ESI-MS/MS method. A selected ion chromatogram for a typical fragment ion (m/z 428) specific to CoA-esters enabled the detection of eight intracellular CoA-esters involved in both primary and secondary metabolisms. The application of this method to bacterial metabolomic study is demonstrated by the profiling of the intracellular CoA-ester pools in the wild-type Streptomyces venezuelae strain producing polyketide antibiotics (methymycin and pikromycin), a polyketide synthase (PKS)-deleted S. venezuelae mutant, and a S. venezuelae mutant expressing the heterologous PKS genes. By quantifying the individual CoA-esterlevel in three different genotypes of the S. venezuela e strain, further insight could be gained into the role of CoA-estersin polyketide biosynthesis. This analytical approach can be extended to the quantification of the size and composition of in vivo CoA-ester pools in various microbes, and can provide a detailed understanding of the relationship between the in vivo CoA-ester pool and the production of pharmaceutically important polyketides. Copyright © 2007 John Wiley & Sons, Ltd. [source] Using Real-time PCR to Discriminate and Quantify the Closely Related Wheat Pathogens Oculimacula yallundae and Oculimacula acuformisJOURNAL OF PHYTOPATHOLOGY, Issue 11-12 2005K. Walsh Abstract Oculimacula yallundae and O. acuformis are the causal agents of eyespot disease of wheat and other cereals. The two fungi react differently to the application of fungicides, but they cannot be distinguished visually and their occurrence is often masked by other less damaging pathogens. Current methods to detect and distinguish Oculimacula species are impractical when testing large numbers of samples. A real-time polymerase chain reaction (PCR) assay, suitable for large-scale testing, was developed and used for quantitative detection and discrimination of O. yallundae and O. acuformis. As the available DNA sequences differ by only a small number of conserved nucleotide polymorphisms, three different methods were investigated to achieve the desired specificity. A combination of mutagenically separated PCR and a shortened primer gave the best specificity for O. yallundae and O. acuformis, respectively. Although the comparison illustrated the practicalities of each method, it was not possible to devise a fixed set of rules for the design of primers to discriminate two closely related sequences; it is assumed that sequence context is an overriding and difficult to predict factor, and that a range of empirical approaches may need to be taken to reach the specificity required. [source] A real-time PCR probe for the quantitative detection of Porphyromonas gingivalisORAL DISEASES, Issue 1 2009DA Apatzidou [source] Validation of a real-time PCR for the quantitative estimation of a G143A mutation in the cytochrome bc1 gene of Pyrenophora teresPEST MANAGEMENT SCIENCE (FORMERLY: PESTICIDE SCIENCE), Issue 3 2007Arash Kianianmomeni Abstract A single nucleotide polymorphism (SNP) in the cytochrome b gene confers resistance to strobilurin fungicides for several fungal pathogens. Therefore, on the basis of a change at amino acid position 143 from glycine to alanine, a real-time PCR assay was established for the quantitative detection of the analogous SNP in the cytochrome b sequence of Pyrenophora teres Drechsler, which causes barley net blotch. Allelic discrimination was achieved by using allele specific primers with artificially mismatched nucleic acid bases and minor groove binding probes. Validation parameters for the lower limits of the working range, namely limits of detection (LOD) and limits of quantification (LOQ), were statistically determined by the variance of calibration data, as well as by the variance of the 100% non-strobilurin-resistant allele DNA sample (blank values). It was found that the detection was limited by the variance of blank values (five in 801 458 copies; 0.0006%), whereas the quantification was limited by the variance of calibration data (37 in 801 458 copies; 0.0046%). The real-time PCR assay was finally used to monitor strobilurin-resistant cytochrome b alleles in barley net blotch field samples, which were already classified in in vivo biotests to be fully sensitive to strobilurins. All signals for strobilurin-resistant cytochrome b alleles were below the LOD, and therefore the results are in total agreement with the phenotypes revealed by biotests. Copyright © 2006 Society of Chemical Industry [source] Enhanced proliferation and efficient transmission of Candidatus Liberibacter asiaticus by adult Diaphorina citri after acquisition feeding in the nymphal stageANNALS OF APPLIED BIOLOGY, Issue 1 2009H. Inoue Abstract We carried out a quantitative detection of Candidatus Liberibacter asiaticus, the bacterium associated with the disease of huanglongbing, in the vector psyllid Diaphorina citri by using a TaqMan real-time PCR assay. The concentration of the bacterium was monitored at 5-day intervals for a period of 20 days after psyllids were exposed as fifth instar nymphs or adults to a Ca. L. asiaticus-infected plant for an acquisition access period of 24 h. When adults fed on Ca. L. asiaticus-infected plant, the concentration of the bacterium did not increase significantly and the pathogen was not transmitted to any citrus seedlings. In contrast, when psyllids fed on infected plant as nymphs, the concentration of the pathogen significantly increased by 25-, 360- and 130-fold from the initial acquisition day to 10, 15 and 20 days, respectively. Additionally, the pathogen was successfully transmitted to 67% of citrus seedlings by emerging adults. Our data suggested that multiplication of the bacterium into the psyllids is essential for an efficient transmission and show that it is difficult for adults to transmit the pathogen unless they acquire it as nymphs. [source] | |||||||||||||||||||