Objective: To compare the sensitivity and reproducibility of Allglo and TaqMan probe in the detection of simian immunodeficiency virus (SIV) using fluorescence quantitative RT-PCR (QPCR).
Methods: The reference sample of SIV was diluted to 6 gradient concentrations; at each concentration 12 samples were tested to analyze the variations within batches, and each sample was tested for 12 times for analysis of variations between batches by QRT-PCR using TaqMan probe and Allglo probe. The results of QPCR using the two probes were analyzed with ABI7300 PCR system software.
Results: In QPCR using TaqMan and Allglo probe, the lower limit of sensitivity for SIV detection was both 50 copies/mL. Assessment of the reproducibility of the tests showed that the maximum and minimum coefficients of variation between batches were 0.63% and 0.33% with Allglo probe, respectively, as compared with 1.33% and 0.2% with TaqMan probe. The maximum and minimum coefficients of inter-batch variation was 1.77% and 0.95% with Allglo probe, respectively, as compared with 1.86% and 1.03% with TaqMan probe.
Conclusion: Allglo probe shows a better performance then TaqMan probe in detection of SIV QPCR.