Figure 4

<p><b>A)</b> Gel electrophoresis of the conventional PCR amplification products showing the undigested 384-bp OPV amplicon DNA ladder (lane 1); negative controls (lanes 2–3); human specimens from 13 patients of the Bena Tshiadi healthcare district (lanes 4–13 and 15–18), and from 1 patient of the Yangala healthcare district (lane 14). <b>B)</b><i>BsrGI</i> digestion profile of 384-bp MPXV amplicons: banding patterns with 210- and 174-bp fragments.</p

Blotting papers showing six 6 spots with dried liquid exudate from pustules collected in one patient.

<p>One spot (arrow) was removed and processed for DNA-based identification of causative agents of the rash illness.</p

DNA-based identification of causative agents of rash illness outbreaks (2008 and 2009) in West Kasai province.

<p>Legend: Patients originated from the following healthcare districts: Bena Tshiadi, 1–13; Yangala, 14–22; Ndesha, 23–25.</p

Map of West Kasai Province.

<p>Areas affected by rash illness outbreaks are highlighted as follows: ▴: Bena Tshiadi healthcare zone (13 patients in 7 different villages). All cases were confirmed as monkeypox cases. Hourglass Symbol: Yangala healthcare zone (8 patients clustered in Tshikongo village). All were confirmed as varicella cases.♦: Ndesha healthcare zone in the outskirts of Kananga (3 patients clustered in Lubuyi village, several kilometers north of Kananga city). All were confirmed as varicella cases.</p

Multiple alignment of 14 kD DNA targets and design of primers and probes for the qPCR assay.

<p>Primers for qPCR and for the conventional PCR are shown in plain boxes whereas the pan-orthopoxvirus and specific variola virus probes are highlighted in a dashed boxes. The monkepoxvirus specific SNP (C→T) is arrowed. The pyrosequencing probe is indicated by a plain arrow.</p