Table 1.
Characteristics of samples.
Table 2.
Sequence of primers and probes designed for the duplex real-time quantitative PCR.
Fig 1.
Specificity of the primers designed for P. jirovecii mtSSU and T. gondii Rep-529 amplification.
The duplex PCR method showed no cross-reaction to Mycobacterium tuberculosis, alpha hemolytic Streptococcus, Neisseria, Mycoplasma or Klebsiella pneumoniae.
Fig 2.
Quantitative correlation between gene copy number and threshold cycle of the duplex qPCR assay.
(A) P. jirovecii mtSSU plasmid was serially diluted from 101 to 108 copies/reaction and subjected to qPCR. (B) Linear regression of Cq vs. lg copy number of mtSSU plasmid using singleplex qPCR and duplex qPCR. (C) T. gondii Rep-529 plasmid was serially diluted from 101 to 108 copies/reaction and subjected to qPCR. (D) Linear regression of Cq vs. lg copy number of Rep-529 plasmid using singleplex qPCR and duplex qPCR. ΔRn = Rn (normalized reporter)-baseline. Ct, Cycle threshold.
Table 3.
The duplex qPCR results for 227 clinical samples.
Fig 3.
The detected P. jirovecii (A) and T. gondii (B) DNA copy number comparison between samples from sputum and BALF.
Fig 4.
Detection of P. jirovecii and T. gondii infection using duplex qPCR assay on clinical samples from 113 lung infection patients.
(A) Number of P. jirovecii, P. jirovecii + T. gondii and T. gondii patients detected by the duplex qPCR. (B) P. jirovecii and T. gondii positive patients detected with duplex qPCR in infant and elder groups. (C) DNA load of P. jirovecii and T. gondii detected in lung infection patients.