A fluorogenic probe hydrolysis (TaqMan) PCR assay for African swine fever virus (ASFV) was developed and evaluated in experimentally infected swine. This sensitive and specific one-step single-tube assay, which can be performed in 2 h or less, detected viral DNA in tonsil scraping samples 2 to 4 days prior to onset of clinical disease. Thus, the assay would have application for preclinical diag...

Xylella fastidiosa is a fastidious Gram-negative bacterium that associated with several important plant diseases, and regulated as quarantine pest in many countries where strategies are implemented to prevent its introduction spread. To enact efficient measures, effective early detection of the pathogen essential, especially because global trade goods increases risks alien pathogens. this study...

A Real Time-PCR method based on TaqMan technology for the 14 identification of Scomber Scombrus has been developed. A system of specific 15 primers and a Minor Groove Binding (MGB) TaqMan probe based on 16 sequences of the mitochondrial cytochrome b region was designed. The 17 method was successfully tested in 81 specimens of Scomber scombrus and 18 related species and validated in 26 different...

To establish an accurate, rapid, and a quantifiable method for the detection of Riemerella anatipestifer infection, a widespread infectious disease in birds, we developed a TaqMan-based real-time PCR assay by using DtxR gene-specific primers and a TaqMan probe. The standard curve established with a linear correlation (R2) of 0.998 and efficiency of 99% between the Ct value and the logarithm of ...

The development of microfluidic tools for high-throughput nucleic acid analysis has become a burgeoning area of research in the post-genome era. Here, we have developed a microfluidic chip to perform 72 parallel 450-pL RT-PCRs. We took advantage of Taqman hydrolysis probe chemistry to detect RNA templates as low as 34 copies. The device and method presented here may enable highly parallel singl...

We have developed a real-time quantitative PCR assay to measure the concentration of DNA extracted in forensic biological traces. Its rapid and sensitive results were obtained in less than 4 hours. This real-time quantification can be performed on the ABI Prism 7700 Sequence Detector using the fluorogenic probe in a TaqMan assay. We suggest a protocol that will be able to detect and quantitate ...

The article describes a new technology for real-time polymerase chain reaction (PCR) detection of nucleic acids. Similar to Taqman, this new method, named Snake, utilizes the 5'-nuclease activity of Thermus aquaticus (Taq) DNA polymerase that cleaves dual-labeled Förster resonance energy transfer (FRET) probes and generates a fluorescent signal during PCR. However, the mechanism of the probe cl...

A multiplex Taqman-based real-time reverse transcription (RT) polymerase chain reaction (PCR) assay was developed to identify potential severe strains of Citrus tristeza virus (CTV) and separate genotypes that react with the monoclonal antibody MCA13. Three strain-specific probes were developed using intergene sequences between the major and minor coat protein genes (CPi) in a multiplex reactio...

AIM To rapidly quantify hepatitis B virus (HBV) DNA by real-time PCR using efficient TaqMan probe and extraction methods of virus DNA. METHODS Three standards were prepared by cloning PCR products which targeted S, C and X region of HBV genome into pGEM-T vector respectively. A pair of primers and matched TaqMan probe were selected by comparing the copy number and the Ct values of HBV serum s...

Scorpion primers can be used to detect PCR products in homogeneous solution. Their structure promotes a unimolecular probing mechanism. We compare their performance with that of the same probe sequence forced to act in a bimolecular manner. The data suggest that Scorpions indeed probe by a unimolecular mechanism which is faster and more efficient than the bimolecular mechanism. This mechanism i...