Figure 4.

NAT-based assessment of non-formalin fixed placental, membrane, and/or umbilical cord tissue for ZIKV demonstrates detectable viral RNA in congenital Zika syndrome-affected cases, but not in the unaffected case. (A) ZIKV NAT in placental, membrane, and cord specimens. ZIKV was detected by NAT in all placental specimens in Cases 1, 2, and 3 (congenital Zika syndrome-affected) but not Case 4 (unaffected). ZIKV was detected in the membranes and cord of Cases 2 and 3 by NAT, but not Cases 1 nor 4. Heat killed conditioned media from Vero cells, with actively replicating clinical isolate ZIKV (20), served as a positive control (ZIKV+), and water as a negative control (H₂O). (B) Quantitative bioanalysis band chromatograms from NAT. Quantitation of ZIKV (74 nt) in relative log measures by NAT was determined employing the Agilent BioAnalyzer. (C) Absence of detectable copy number or potentially pathologic SNP nor exon variants by CMA and whole exome sequencing. A novel de novo heterozygous c.3667G > C (p.V1223L) variant in the MED12 gene (located on ChrX:70349255) of unknown clinical significance (VUS) was found. No advanced genomic testing was performed with Case 4 secondary to absence of ultrasound findings and normal neonatal and infant clinical examinations.