Table 1.
Primer-probe information for Streptococcus pneumoniae, Haemophilus influenzae, and Moraxella catarrhalis.
Fig 1.
Sensitivity analysis of the multiplex fluorescence PCR method for three fastidious bacteria.
A: 1.5 × 10^6 Copies/ml, B: 1.5 × 10^5 Copies/ml, C: 1.5 × 10^4 Copies/ml, D: 1.5 × 10^3 Copies/ml, E: 500 Copies/ml, F: 200 Copies/ml, G: 100 Copies/ml, H: 50 Copies/ml, I: 20 Copies/ml, J: 10 Copies/ml. Limit of detection for Streptococcus pneumoniae: 100 Copies/ml. Limit of detection for Haemophilus influenzae: 20 Copies/ml. Limit of detection for Moraxella catarrhalis: 50 Copies/ml.
Table 2.
The information of the 173 participants.
Table 3.
Distribution of three pathogenic bacteria in clinical specimens detected by bacterial culture, multiplex fluorescence PCR, and tNGS methods.
Table 4.
Positive rates of three pathogenic bacteria by bacterial culture, bacterial culture + tNGS, and multiplex fluorescence PCR methods in clinical specimens.
Table 5.
Comparison of multiplex fluorescence PCR results with bacterial culture + tNGS (reference method).
Table 6.
Comparative Table (Culture vs. Multiplex PCR).
Table 7.
χ2 Test Results for Culture vs. Multiplex PCR.
Fig 2.
Amplification plots of representative positive and negative clinical specimens using the multiplex fluorescent PCR assay.
Red, green, and blue fluorescence curves represent Streptococcus pneumoniae (SP), Haemophilus influenzae (HI), and Moraxella catarrhalis (MC), respectively. Panel A shows an SP-only positive sample (red curve only), Panel B an HI-only positive sample (green curve only), Panel C an MC-only positive sample (blue curve only), Panel D a negative control (no amplification), and Panel E a triple-positive sample with red, green, and blue curves.
Fig 3.
Overview of Multiplex PCR Workflow and Results (by FigDraw).
This figure was created using FigDraw (www.figdraw.com). The images and elements are original and have been authorized for use. This figure was drawn by the first author, Jin Chao Shi.