Quantitative real-time PCR assay for rapid detection of plant and human pathogenic Macrophomina phaseolina from field and environmental samples

Babu Bandamaravuri Kishore; Mesapogu Sukumar; Sharma Anu; Somasani Saida Reddy; Arora Dilip K. .

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Quantitative real-time PCR assay for rapid detection of plant and human pathogenic Macrophomina phaseolina from field and environmental samples

机译：实时荧光定量PCR检测法可从田间和环境样品中快速检测植物和人类致病性菜豆

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A real-time qPCR assay was developed to detect and quantify Macrophomina phaseolina abundance in rhizosphere soil and plant tissue. Both TaqMan and SYBR green techniques were targeted on similar to 1 kb sequence characterized amplified region (SCAR) of M. phaseolina and two sets of specific primers were designed for SYBR green (MpSyK) and TaqMan (MpTqK) assays. No cross-hybridization and no fluorescent signal exceeding the baseline threshold was observed in TaqMan and SYBR green assays, respectively. The minimum detection limit or sensitivity of TaqMan assay was 30 fg/mu L of M. phaseolina DNA and limit of quantification of M phaseolina viable population was estimated as 0.66 x 10(5) CFU/g soil(-1) equivalent to 10 pg/mu L of target DNA. This is the first report which demonstrated real-time qPCR assays with greater specificity and sensitivity to detect M. phaseolina population in soil and plant materials.

机译：开发了一种实时定量PCR检测试剂盒，以检测和定量根际土壤和植物组织中的菜豆中巨噬藻的丰度。 TaqMan和SYBR green技术都针对相近的嗜盐支原体的1 kb序列特征扩增区域（SCAR），设计了两组特异性引物用于SYBR green（MpSyK）和TaqMan（MpTqK）分析。在TaqMan和SYBR green检测中分别未观察到交叉杂交和荧光信号超过基线阈值。 TaqMan分析的最低检测限或灵敏度为30 fg /μL菜豆分枝杆菌DNA，M菜豆分枝杆菌存活种群的定量限估计为0.66 x 10（5）CFU / g土壤（-1）相当于10 pg /μL的目标DNA。这是第一份报告，该报告展示了实时qPCR分析，具有更高的特异性和灵敏度，可检测土壤和植物材料中的菜豆分枝杆菌种群。

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《Mycologia》 |2011年第3期|共8页
作者
Babu Bandamaravuri Kishore; Mesapogu Sukumar; Sharma Anu; Somasani Saida Reddy; Arora Dilip K. .;
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中图分类侵（传）染性病害;
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primersSequence DataGU081134: GenBank; EMBL; DDJB; nucleotide sequence; GU081135: GenBank; EMBL; DDJB; nucleotide sequence; GU081142: GenBank; EMBL; DDJB; nucleotide sequence; GU081136: GenBank; EMBL; DDJB; nucleotide sequence; GU081137: GenBank; EMBL; DDJB; nucleotide sequence; GU081138: GenBank; EMBL; DDJB; nucleotide sequence; GU081139: GenBank; EMBL; DDJB; nucleotide sequence; GU081140: GenBank; EMBL; DDJB; nucleotide sequence; GU081141: GenBank; EMBL; DDJB; nucleotide sequence; detection limit; rhizosphere soil; environmental sample; cross-hybridization; sequence characterized amplified region; vegetative population; plant tissue;

机译：序列数据GU081134：GenBank;EMBL;DDJB;核苷酸序列;GU081135：GenBank;EMBL;DDJB;核苷酸序列;GU081142：GenBank;EMBL;DDJB;核苷酸序列;GU081136：GenBank;EMBL;DDJB;核苷酸序列;GU081137：GenBank;EMBL;DDJB;核苷酸序列;GU081137：GenBank;EMBL;DDBL;DDJB;核苷酸序列;GU081138：GenBank;EMBL;DDJB;核苷酸序列;GU081139：GenBank;EMBL;DDJB;核苷酸序列;GU081140：GenBank;EMBL;DDJB;核苷酸序列;GU081141：GenBank;EMBL;DDJB;核苷酸序列;检出限;根际土壤;环境样品;交叉杂交;序列鉴定的扩增区;营养种群;植物组织;

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专利

1. Quantitative real-time PCR assay for rapid detection of plant and human pathogenic Macrophomina phaseolina from field and environmental samples [J] . Babu Bandamaravuri Kishore, Mesapogu Sukumar, Sharma Anu, Mycologia . 2011,第3期

机译：实时荧光定量PCR检测法可从田间和环境样品中快速检测植物和人类致病性菜豆
2. Quantitative detection and identification of Naegleria spp. in various environmental water samples using real-time quantitative PCR assay. [J] . Po-Min Kao, Min-Che Tung, Bing-Mu Hsu, Parasitology Research . 2013,第4期

机译：定量检测和鉴定Naegleria spp。使用实时定量PCR分析法在各种环境水样品中进行分析。
3. Development and validation of a real-time quantitative PCR assay for rapid identification of Bacillus anthracis in environmental samples [J] . Irenge L.M., Durant J.-F., Tomaso H., Applied Microbiology and Biotechnology . 2011,第5期

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4. Rapid Detection of Pathogenic Aspergillus species in Avian Samples by Real-time PCR Assay: A Preliminary Report [C] . Bob Dahlhausen, Rob Abbott, Pam VanOverloop Annual Conference of the Association of Avian Veterinarians . 2004

机译：通过实时PCR测定快速检测禽样品中禽样品中的致病性曲霉属物种：初步报告
5. Rapid and sensitive detection of aflatoxin in animal feeds and food grains using immunomagnetic bead based recovery and real-time immuno quantitative PCR (RT-iqPCR) assay. [D] . Babu, Dinesh. 2010

机译：使用基于免疫磁珠的回收率和实时免疫定量PCR（RT-iqPCR）分析快速，灵敏地检测动物饲料和粮食中的黄曲霉毒素。
6. Development of a Multiplex Real-Time PCR Assay for Rapid Detection of Tigecycline Resistance Gene tet(X) Variants from Bacterial Fecal and Environmental Samples [O] . Yulin Fu, Dejun Liu, Huangwei Song, 2020

机译：开发多重实时PCR测定用于快速检测从细菌粪便环境样品中快速检测Tigeccline抗性基因TET（X）变体
7. Rapid quantitative detection of chytridiomycosis (Batrachochytrium dendrobatidis) in amphibian samples using real-time Taqman PCR assay [O] . Boyle, D. G., Boyle, D. B., Olsen, V., 2004

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8. Evaluation and Validation of a Real-Time PCR Assay for Detection and Quantitation of Human Adenovirus 14 from Clinical Samples [R] . Metzgar, D., Skochko, G., Gibbins, C., 2009

机译：用于从临床样品中检测和定量人腺病毒14的实时pCR测定的评估和验证

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Quantitative real-time PCR assay for rapid detection of plant and human pathogenic Macrophomina phaseolina from field and environmental samples

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