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Microbial DNA qPCR Assays

采用real-time PCR技术进行微生物检测
  • 经实验验证,具有高度灵敏性和特异性
  • 可检测微生物物种、毒力因子基因、或抗生素耐药性基因
  • 简单便利的qPCR工作流程
Microbial DNA qPCR Assays含有两种PCR引物(每种10 µM)、以及一种5'水解探针(5 µM)用于定量real-time PCR。Microbial DNA qPCR Assays采用经实验验证的专有技术,其PCR效率高,扩增效果准确、稳定。所有的Microbial DNA qPCR Assay都经过严格的实验验证,PCR反应效率高。该产品搭配Microbial qPCR Mastermixes和Microbial DNA-Free Water使用,可确保PCR的高效率。
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You can search for the following terms:
  • Entrez Gene IDs (e.g., 835)
  • RefSeq IDs (e.g., NM_032983, NP_116765)
  • Gene symbols (e.g., CASP2)
  • Cat. no. (e.g., SI00299551, QT01342509)
  • Sanger ID or Accession (e.g., hsa-let-7b, MI0000063)
  • CpG loci identification numbers (CG#) (e.g., CG17753661)

Do not enter species information in the Search box. Use the drop-down list to select your species of interest.
When searching for miRNAs, do not omit the hyphens. Use hyphens or spaces (e.g., search for hsa-let-7b or hsa let 7b. Do not search for hsalet7b).

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产品 Product no. 货号 目录价:
Microbial DNA qPCR Assay
One 100 µl tube Microbial DNA qPCR Assay, one 1.35 ml tube Microbial qPCR Mastermix
330025 Varies Enter your targets in search box above.
Custom Microbial DNA qPCR Assay
One 100 µl tube newly designed Custom Microbial DNA qPCR Assay, one 1.35 ml tube Microbial qPCR Mastermix
330061 Varies Enter your targets in search box above.
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Microbial DNA qPCR Assays适用于分子生物学应用。该产品不适用于疾病的诊断、预防或治疗。


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Limit of detection versus lower limit of quantification.
This chart demonstrates the difference between the limit of detection (LOD) and the lower limit of quantification (LLOQ). The LOD is defined as the lowest concentration at which 95% of the positive samples are detected, whereas the LLOQ is the lowest concentration that falls within the linear range of a standard curve. LOD depends upon the precision of the assay, and requires at least 40 replicates for determination of a positive sample. For the Microbial DNA qPCR Assays, LLOQ is sufficient to determine assay sensitivity.
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The lower limit of quantification (LLOQ) for all Microbial DNA qPCR Assays reveals high sensitivity.
This chart shows the distribution of LLOQ for all Microbial DNA qPCR Assays. 93% of all Microbial DNA qPCR Assays have a LLOQ of <100 gene copies.
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The lower limit of quantification (LLOQ) for microbial identification Microbial DNA qPCR Assays reveals high sensitivity.
This chart shows the distribution of LLOQ for microbial identification Microbial DNA qPCR Assays. 92% of all microbial identification assays have a LLOQ of <100 gene copies.
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The lower limit of quantification (LLOQ) for virulence factor gene detection Microbial DNA qPCR Assays reveals high sensitivity.
This chart shows the distribution of LLOQ for Microbial DNA qPCR Assays for virulence factor gene detection. 97% of all virulence factor gene assays have a LLOQ of <100 gene copies.
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The lower limit of quantification (LLOQ) for antibiotic resistance gene detection Microbial DNA qPCR Assays reveals high sensitivity.
This chart shows the distribution of LLOQ for Microbial DNA qPCR Assays for antibiotic resistance gene detection. 95% of all antibiotic resistance gene assays have a LLOQ of <100 gene copies.
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Microbial DNA qPCR Assays are highly specific.
To determine the specificity of Microbial DNA qPCR Assays, each assay was tested against 119 genomic DNA samples from different bacteria and fungi. To facilitate testing, genomic DNA from different microbial species was pooled (10 different genomic DNA samples per pool) and each assay was tested against the different pools. None of the pools contained DNA from the same genus, to facilitate identification of cross-reacting species. Each pool contained the equivalent of 2000 genome copies for each microbial species. In addition, each assay was tested against human, mouse, and rat genomic DNA. A representative example for Streptococcus pyogenes is shown. The assay for Streptococcus pyogenes gave a CT of 26.9 and 26.6 for the Staphylococcus/Streptococcus pool and complete pool [A]. Both pools contained genomic DNA for Streptococcus pyogenes. To determine which genomic DNA was detected by the Streptococcus pyogenes assay, each individual genomic DNA comprising the Staphylococcus/Streptococcus pool was tested separately [B]. Only Streptococcus pyogenes genomic DNA gave an acceptable CT call (26.8) while the others gave a CT>35. Most of the assays were specific as they did not detect unintended targets. For assays that detected other species, the list of detected targets along with in silico predictions are given in the specifications sheet.
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Microbial DNA qPCR Assays display high sensitivity even in complex metagenomic samples.
To ensure that Microbial DNA qPCR Assays performed comparably in a complex sample, where there may be up to a thousand different microbial species, each assay was tested using stool, tooth plaque, and sputum samples. For each sample, synthetic template targets were spiked in and the CT was compared to synthetic template alone. PCR was performed using several sample types, which included pooled synthetic template targets alone, stool, stool plus pooled synthetic template targets, plaque, plaque plus pooled synthetic template targets, sputum, and sputum plus pooled synthetic template targets. If the CT<35 in stool, plaque, or sputum samples alone, then ΔCT was calculated (i.e., CTstool – CTpooled synthetic template targets). This calculation was performed for all the assays. For each assay, the ΔCT<3, indicating that a complex metagenomic background does not affect the performance of each Microbial DNA qPCR Assay.
Performance
最低定量阈限(LLOQ)
LLOQ是指在标准曲线线性范围内的最低模板浓度(参见Limit of detection versus lower limit of quantification)。所有Microbial DNA qPCR Assays中的93%具有低至100个基因拷贝的LLOQ(参见The LLOQ for all Microbial DNA qPCR Assays reveals high sensitivity)。该系列产品中的92%微生物检测产品能达到这一LLOQ,95%的毒力因子检测产品和97%的抗生素耐药性基因检测产品能达到这一LLOQ(参见The LLOQ for microbial identification Microbial DNA qPCR Assays reveals high sensitivityThe LLOQ for virulence factor genes Microbial DNA qPCR Assays reveals high sensitivityThe LLOQ for antibiotic resistance genes Microbial DNA qPCR Assays reveals high sensitivity)。

特异性
所有Microbial DNA qPCR Assay都经过严格检验,确保能够灵敏检测某一物种或基因(参见Microbial DNA qPCR Assays are highly specific)。对于能够检测一种以上五种或基因的产品,产品说明页上附有电脑预测可检测靶标的列表。

即便样本中存在多种物种,该试剂盒仍有极高的检测特异性,此类样本包括粪便、痰和菌斑(参见Microbial DNA qPCR Assays display high sensitivity even in complex metagenomic samples)。
Principle
Microbial DNA qPCR Assays在进行物种检测时,可检测细菌的16S rRNA基因和真菌核糖体rRNA基因序列;采用PCR扩增引物和水解探针,检测毒力因子基因和抗生素耐药性基因。
Procedure
Microbial DNA qPCR Assay的实验流程简单,能够在各种real-time PCR仪上使用。使用相应的QIAamp试剂盒从样本中分离DNA,然后使用与PCR仪相应的Supplemental Microbial qPCR Mastermix构建PCR反应体系。为每种样本构建4个独立的PCR反应体系,包括Positive PCR Control、No Template Control、Microbial DNA Positive Control以及Microbial DNA qPCR Assay。进行real-time PCR,然后使用在线数据分析软件或Excel模板文件进行数据分析。
Applications
Microbial DNA qPCR Assay高度适用于细菌或真菌物种检测,以及微生物的抗生素耐药性基因或毒力因子检测,适用样本包括粪便、痰、阴道拭子、污水等等。

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产品介绍与指南 (2)
Microbial product configurator guide
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For application-specific microbial identification and profiling by real-time PCR
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常见问题 (20)
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试剂盒操作手册 (1)
New version – For real-time PCR-based profiling/detection of microbial species, antibiotic resistance genes or virulence factor genes
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