Supplementary material 1 Real-time quantitative PCR: Sequences of the primers and TaqMan probe for ADAMTS1, 4, 5, 8, 9 and 15 were designed using Primer Express software (Applied Biosystems, Foster City, CA): forward primer 5’-CCATCCCAAGAGTATCACATGTCT-3’, reverse primer 5’-CACTATGACACAGCAATTCTTTTCAC-3’ and TaqMan probe 5’-FAM-CCCACACAAGTCCTGTC-MGB-3 for ADAMTS1; forward primer 5’-TCACTGACTTCCTG...

Insulated isothermal PCR (iiPCR), established on the basis of Ralyeigh-Bénard convection, is a rapid and low-cost platform for nucleic acid amplification. However, the method used for signal detection, namely gel electrophoresis, has limited the application of iiPCR. In this study, TaqMan probe-based iiPCR system was developed to obviate the need of post-amplification processing. This system in...

Background and Aims: Newcastle disease virus (NDV) is an avian paramyxovirus (A-PMV 1) and one of the major pathogens in poultries. Vaccination is intended to control the disease, nevertheless this virus is a growing threat to the poultry industry. So, early detection of the virus can prevent the spread of illness and avoid huge economic losses. Towards this goal, in this research, we develop...

Ongoing evolution of viral pathogens is a significant issue in diagnostic virology employing TaqMan qPCR/RT-qPCR. Specific concerns are related to false negativity due to probe binding failure. One option for compensating for such deficiency is to integrate a second identically labelled probe in the assay. However, how this alteration influences the reaction parameters has not been comprehensiv...

TaqMan probe (dual-labeled DNA probe)-based real-time detection, one of the most sensitive and specific fluorescent detection methods, has been widely utilized in conjunction with polymerase chain reaction (PCR). Helicase-dependent amplification (HDA) is an isothermal amplification technology that has a similar reaction scheme to PCR, but replaces thermocycling with a helicase capable of unwind...

background and aims: newcastle disease virus (ndv) is an avian paramyxovirus (a-pmv 1) and one of the major pathogens in poultries. vaccination is intended to control the disease, nevertheless this virus is a growing threat to the poultry industry. so, early detection of the virus can prevent the spread of illness and avoid huge economic losses. towards this goal, in this research, we developed...

We investigated three probe design strategies used in quantitative polymerase chain reaction (PCR) for sensitivity in detection of the PCR amplicon. A plasmid with a 120-bp insert served as the DNA template. The probes were TaqMan, conventional molecular beacon (MB), and shared-stem molecular beacon (ATssMB and GCssMB). A shared-stem beacon probe combines the properties of a TaqMan probe and a ...

Several methods for the detection of RNA have been developed over time. For small RNA detection, a stem-loop reverse primer-based protocol relying on TaqMan RT-PCR has been described. This protocol requires an individual specific TaqMan probe for each target RNA and, hence, is highly cost-intensive for experiments with small sample sizes or large numbers of different samples. We describe a univ...

Disease outbreaks caused by eastern equine encephalitis virus (EEEV; Togaviridae, Alphavirus) may be prevented by implementing effective surveillance and intervention strategies directed against the mosquito vector. Methods for EEEV detection in mosquitoes include a real-time reverse transcriptase PCR technique (TaqMan assay), but we report its failure to detect variants isolated in Connecticut...