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Figure 2 | BMC Microbiology

Figure 2

From: Sensitive multiplex PCR assay to differentiate Lyme spirochetes and emerging pathogens Anaplasma phagocytophilum and Babesia microti

Figure 2

Molecular beacons can detect B. burgdorferi between 1 and 106in a duplex assay, when human DNA was also included. Amplification plots of recA and Actin A1 genes in PCR assays to estimate quantities of B. burgdorferi (A) and human (C) DNA are shown. Human DNA (containing 105 Actin A1 gene copies) spiked with ten-fold dilutions of B. burgdorferi strain N40 ranging from 1 to 106 were used in the PCR assays containing both RecA3 and ACTA1 molecular beacons. Sensitivity and specificity of the detection system is indicated by the ability of RecA3 and ACTA1 molecular beacons to quantitatively detect the amplicons from both the recA and the ACTA1 genes in the same PCR assay tubes. A high coefficient of correlation (r2 = 0.999) between the Ct values and the spirochete number obtained from the standard curve (B) indicates that the molecular beacons can be used effectively to quantify spirochete burden in infected tissues using multiplex assay system.

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