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Gene Synthesis> | As a positive control, the target sequence of the upE assay and flanking nucleotides (c. 400 bp total) were synthesized by a commercial source (Genscript, Piscataway, NJ, USA) and cloned into plasmid pUC57. One-step rRT-PCR reactions were prepared using the AgPath-ID One-Step RT-PCR kit (Life Technologies) according to the manufacturer‘s instructions with 400 nM forward and reverse primers and 200 nM probe. Cycling parameters were 50°C × 30 min, 95°C × 10 min and 45 cycles of 95°C × 15 s and 58°C × 30 s, with fluorescence data collected during the 60°C annealing/extension step. | Get A Quote |
Hospitalized children < 2 years of age in Amman, Jordan, admitted for fever and/or respiratory symptoms, were tested for Middle East respiratory syndrome coronavirus (MERS-CoV): MERS-CoV by real-time RT-PCR (rRT-PCR). This was a prospective year-round viral surveillance study in children <2 years of age admitted with acute respiratory symptoms and/or fever from March 2010 to September 2012 and enrolled from a government-run hospital, Al-Bashir in Amman, Jordan. Clinical and demographic data, including antibiotic use, were collected. Combined nasal/throat swabs were collected, aliquoted, and frozen at –80°C. Specimen aliquots were shipped to Vanderbilt University and the Centers for Disease Control and ... More