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Table 2 Evaluation of Filtration methods to improve sensitivity and specificity of rt-qPCR assays

From: A real-time, quantitative PCR method using hydrolysis probes for the monitoring of Plasmodium falciparum load in experimentally infected human volunteers

Dilutions (3 × replicates) Hydrolysis probe assay SYBR Green assay
   Crossing point (SD), Quant value (SD)
Filtered 5.0 × 105 23.38 (0.03), 540 000 (9 000) 20.95 (0.68) 850 000 (400 000)
  1.0 × 105 25.36 (0.16) 160 000 (16 000) 24.20 (0.44) 81 000 (26 000)
  2.0 × 104 28.33 (0.17) 27 000 (3000) 26.14 (0.31) 20 000 (4000)
  1.0 × 103 33.74 (0.44) 1000 (300) 32.65 (0.49) 200 (70)
  5.0 × 102 34.64 (0.45) 600 (160) 33.29 (1.24) 160 (130)
  2.5 × 102 37.40 (n/a) 110 (n/a) 37.02 (1.05)a 10 (10)
  1.0 ×02 Not detected   False Positive 20 (10)
Unfiltered 5.0 × 105 24.53 (0.62) 300 000 (94 000) 22.83 (0.46) 200 000 (70 000)
  1.0 × 105 25.46 (0.35) 156 000 (33 000) 24.50 (0.11) 64 000 (4800)
  2.0 × 104 29.15 (0.05) 17 000 (500) 29.85 (0.44) 1500 (500)
  1.0 × 103 33.81 (0.38) 1000 (220) 32.98 (0.16) 160 (20)
  5.0 × 102 35.27 (1.63) 550 (450) 36.38 (0.23)a 15 (3)
  2.5 × 102 38.35 (1.06) 75 (50) 35.63 (0.73)a 30 (10)
  1.0 ×02 Not detected   False Positive 10 (2)
  1. a Indicates nonspecific melting peaks of 79°C obtained by melting curve analysis of replicates tested.
  2. n/a = Not Applicable
  3. SD = Standard Deviation
  4. ND = Not Detected