Detection of parvovirus mRNAs as markers for viral activity in endomyocardial biopsy-based diagnosis of patients with unexplained heart failure

Figure 1. (A) Schematic description of the B19V genome organization and PCR design. Gene locus and primer and probe localization for NS1 and VP1/2 are indicated by numbers representing the nucleotide position. ITR inverted terminal repeat, NS1 non structural protein 1, VP1/2 capsid proteins, P6 P6 promotor, (pA)p polyadenylation site proximal, (pA)d polyadenylation site distal. (B) Representative agarose gel electrophoresis gel blot image of a B19V-VP1/2 DNA and RNA-positive EMB using VP1/2 specific nested-PCR. Amplicon length 173 bps. 1 = DNA-Marker 100 bps; 2 = positive control; 3 = negative control; 4 = PCR after DNA extraction and DNAse treatment; 5 = PCR after RNA extraction, RNAse treatment and RT-PCR; 6 = PCR after DNA extraction; 7 = PCR after RNA extraction and DNAse treatment and RT-PCR. Complete gel blot image of figure (B) was shown in Supplementary Fig. S1. (C) Representative agarose gel electrophoresis gel blot image of 10 EMB samples following VP1/2 specific nested PCR. DNA (first lane) and cDNA (second lane) of each EMB were analysed. Amplicon length 173 bps EMBs 1, 6, 8, 9 and 10 were tested positive for viral DNA and negative for viral RNA. EMBs 3 and 5 were positive for…

Figure 2. (A) B19V genome detection and detection of viral transcription activity in EMBs of patients with unexplained heart failure (N = 576). (B) The group composition of EMBs with detectable active viral transcription (VP1/2-RNA-, NS1-RNA and VP1/2 and NS1-RNA positive samples) was shown in detail. Numbers represent the amount of EMBs.

Figure 3. (A) The number of viral transcripts of NS1 compared to VP1/2. (B) Viral DNA loads in EMBs with active or latent infection. (C) Viral DNA load compared between EMBs with detectable VP1/2-RNA, NS1-RNA or NS1- and VP1/2-RNA expression (ANOVA p = 0.0427). Numbers above the bars represent p-values.