Enhanced detection of measles virus infection following successful development of a real-time reverse transcriptase PCR in respiratory and urine samples using the cepheid SmartCycler®

Pathology ◽  
2015 ◽  
Vol 47 ◽  
pp. S57
Author(s):  
Kyra Y.L. Chua ◽  
Kiran Thapa ◽  
Chaturangi M. Yapa ◽  
Lucy K. Somerville ◽  
Vicky Sheppeard ◽  
...  
Keyword(s):  
Virus Infection ◽  
Reverse Transcriptase ◽  
Real Time ◽  
Measles Virus ◽  
Urine Samples ◽  
Successful Development ◽  
Reverse Transcriptase Pcr

scholarly journals Development of an assay for the detection and quantification of the measles virus nucleoprotein (N) gene using real-time reverse transcriptase PCR

2009 ◽  
Vol 58 (5) ◽  
pp. 638-643 ◽  
Author(s):  
Miho Akiyama ◽  
Hirokazu Kimura ◽  
Hiroyuki Tsukagoshi ◽  
Katsuya Taira ◽  
Katsumi Mizuta ◽  
...  
Keyword(s):  
Reverse Transcriptase ◽  
Real Time ◽  
Measles Virus ◽  
N Gene ◽  
Reverse Transcriptase Pcr ◽  
Quantification Method ◽  
Infected Cells ◽  
Syncytial Virus ◽  
Nucleoprotein N ◽  
Detection And Quantification

We developed a new quantification method for the measles virus (MeV) nucleoprotein (N) gene using real-time reverse transcriptase PCR. This method allowed us to quantify 101–107 copies per reaction (corresponding to 5×10−1–5×105 copies μl−1) of the MeV N gene. We also quantified the MeV N gene from the throat swabs of 22 patients with measles as well as the MeV genotypes A, D3, D5, D9 and H1 in viral suspensions derived from MeV-infected cells. As a result, 3.9×103–5.2×106 copies ml−1 and 7.4×107–2.0×108 copies ml−1 of the MeV genomes (N gene) were detected in the throat swabs and viral suspensions, respectively. No other viruses (enteroviruses, respiratory syncytial virus, human metapneumovirus or mumps virus) were detected in the assay. The results suggest that this method is applicable to the detection and quantification of some genotypes of MeV.

Validation of a Real-Time Reverse Transcriptase–PCR Assay for the Detection of H7 Avian Influenza Virus

2010 ◽  
Vol 5 (s1) ◽  
pp. e148-e149
Author(s):  
Janice Pedersen ◽  
Mary Lea Killian ◽  
Nichole Hines ◽  
Dennis Senne ◽  
Brundaban Panigrahy ◽  
...  
Keyword(s):  
Influenza Virus ◽  
Avian Influenza ◽  
Reverse Transcriptase ◽  
Avian Influenza Virus ◽  
Real Time ◽  
Pcr Assay ◽  
Reverse Transcriptase Pcr

Development of asigDE-Based Real-Time Reverse-Transcriptase PCR for the Detection of ViableSalmonella enterica

2014 ◽  
Vol 11 (7) ◽  
pp. 537-544 ◽  
Author(s):  
Min Zhou ◽  
Jielin Yang ◽  
Xiujuan Zhou ◽  
Bin Liu ◽  
Daixin Liu ◽  
...  
Keyword(s):  
Reverse Transcriptase ◽  
Real Time ◽  
Reverse Transcriptase Pcr

scholarly journals Comparison of Rapid Antigen Test and Real-Time Reverse Transcriptase PCR for Diagnosing Novel Swine Influenza A (H1N1)

2010 ◽  
Vol 13 (3) ◽  
pp. 109 ◽  
Author(s):  
Aerin Kwon ◽  
Jae-Seok Kim ◽  
Han-Sung Kim ◽  
Wonkeun Song ◽  
Ji-Young Park ◽  
...  
Keyword(s):  
Reverse Transcriptase ◽  
Real Time ◽  
Influenza A ◽  
Swine Influenza ◽  
Antigen Test ◽  
Reverse Transcriptase Pcr ◽  
Influenza A H1n1

Novel Approach for Detecting Prohibited Species-Specific Central Nervous System Tissue Contamination in Meat by One-Step Real-Time Reverse Transcriptase PCR

2009 ◽  
Vol 72 (5) ◽  
pp. 1063-1069 ◽  
Author(s):  
M. M. NAGARAJAN ◽  
D. LONGTIN ◽  
C. SIMARD
Keyword(s):  
Central Nervous System ◽  
Nervous System ◽  
Reverse Transcriptase ◽  
Real Time ◽  
Compliance Monitoring ◽  
Reverse Transcriptase Pcr ◽  
Qrt Pcr ◽  
Tissue Contamination ◽  
One Step ◽  
Species Specific

The dissemination of prohibited species-specific central nervous system (CNS) tissue contamination in meat must be tracked to mitigate human health risk associated with bovine spongiform encephalopathy. The efficiency of compliance monitoring and risk control measures taken by concerned regulatory authorities at meat production facilities to avoid such contamination depends on the ability to detect CNS tissue with a reliable and adequately sensitive quantitative method. A rapid and convenient one-step real-time quantitative reverse transcriptase PCR (qRT-PCR) assay was developed based on the absolute quantification of glial fibrillary acidic protein (GFAP) mRNA as a marker for CNS tissue contamination in meat. The GFAP RNA quantity corresponding to a percentage of CNS tissue in artificially spiked meat was determined using an appropriate in vitro transcribed target GFAP RNA as a calibration standard in the assay. The assay had a linear dynamic range of 102 to 109 copies of target RNA and was able to detect 0.01% CNS contamination in meat. Further evaluation consisted of an analysis of 272 random meat cuts from carcasses and 109 ground meat samples received from a federally inspected abattoir and two meat processing facilities, respectively, over a 5-month period. The analyzed samples were all negative for CNS tissue contamination at an arbitrarily set lower threshold of 0.025%. Overall, the newly developed one-step qRT-PCR may be useful as an objective quantitative compliance monitoring tool and for setting an acceptable low tolerance threshold for such contamination in meat.

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