Real-time PCR assay

M. Panning, M. Asper, S. Kramme, H. Schmitz and C. Drosten, Rapid detection and differentiation of human pathogenic orthopox viruses by a fluorescence resonance energy transfer real-time PCR assay, Clin. Chem., 50 (2004) 702-708. 

Mule, G., Susca, A., Logrieco, A., Stea, G., and Visconti, A. (2006). Development of a quantitative real-time PCR assay for the detection of Aspergillus carbonarius in grapes. Int. J. Food Microbiol. Ill, S28-S34. 

Jothikumar N, Griffiths MW (2002) Rapid detection of Escherichia coli 0157 H7 with multiplex real-time PCR assays. Appl Environ Microbiol 68 3169-3171... 

Ikewaki L Ohtsuka E, Kawano R, Ogata M, Kikuchi H, Nasu M. Real-time PCR assay compared to nested PCR and antigenemia assays for detecting cytomegalovirus reactivation in adult T-ceH leukemia-lymphoma patients. J CUn Microbiol 2003 41 4382-7. 

Gamier, L., Gaudin, J.-C., Bensadoun, P., Rebillat, I., and Morel, Y. 2009. Real-time PCR assay for detection of anew simulant for poxvirus biothreat agents. Appl. Environ. Microbiol. 75 1614-1620. 

Beqaj SH, Flesher R, Walker GR, et al. Use of the real-time PCR assay in conjunction with MagNA Pure for the detection of mycobacterial DNA from fixed specimens. Diagn Mol Pathol. 2007 16 169-173. 

Every reaction well in the real-time PCR assay plate received 45 pL of the master-mix along with 5 pL of the PLA reaction. Every PLA reaction was assayed in triplicate (three separate additions of 5 pL). 

In addition to the PLA samples, 5 pL of a full length template stock was mixed with 45 pL of master-mix and served as the positive control on the real-time PCR assay plate. 

Once the real-time PCR assay plate was prepared, the samples were mixed carefully with a multi-channel pipette, the wells were sealed with a Microamp full plate cover seal (ABI, Foster City, CA), and the plate was centrifuged at 1,5 OOrpm (-238 x g) at 4°C for 1 min. 

Bowers, H.A. et aL, Development of real-time PCR assays for rapid detection of Pfiesteria piscicida and related dinoflagellates, Appl. Environ. Microbiol. 66, 4641, 2000. 

Degradation of the DNA In most real-time PCR assays, fragments of 60 to 100 bp are amplified. For degraded DNA these short fragments can be overrepresented. This... 

Scaravelli E, Brohee M, Marchelli R, van Hengel AJ (2008). Development of three real-time PCR assays to detect peanut allergen residue in processed food products. Fur. Food Res. Technol, 227(3) 857-869. 

Molecular biological techniques use the presence of species-specific DNA-sequences for detection. For lupin, more than 2500 expressed sequence tags are available in public databases. Quite recently, a hybridization probe-based real-time PCR assay for the detection of lupin DNA in foods was developed (Demmel... 

Debretsion, A. et al.. Real-time PCR assay for rapid detection and quantification of Campylobacter jejuni on chicken rinses from poultry processing plant. Mol. Cell. Probe, 21,177, 2007. 

Gyarmati, P. et al.. Universal detection of Hepatitis E virus by two real-time PCR assays TaqMan and Primer-Probe Energy Transfer, J. Virol Methods, 146, 226, 2007. 

Bell, A.S. and Ranford-Cartwright, L.C. (2004) A real-time PCR assay for quantifying Plasmodium falciparum infections in the mosquito vector. Int. J. Parasitol. 34, 795-802. 

Phister, T. G., Mills, D. A. (2003). Real-time PCR assay for detection and enumeration of Dekkera bruxellensis in wine. Applied and Environmental Microbiology, 69,... 

Yoshida, A., Suzuki, N., Nakano, Y. et al. 2003. Development of 5 fluorogenic nuclease-based real-time PCR assay for quantitative detection of Actino bacillus actinomy cete mco mitans and Porphyromonas gingi-valis. J. Clin. Microbiol. 41 863-866. 

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