A Case Report of a Middle East Respiratory Syndrome Survivor with Kidney Biopsy Results

Cha, Ran-hui; Yang, Seung Hee; Moon, Kyung Chul; Joh, Joon-Sung; Lee, Ji Yeon; Shin, Hyoung-Shik; Kim, Dong Ki; Kim, Yon Su

Confocal microscopy was performed with an LSM510 META confocal microscope (Carl Zeiss, Jena, Germany). For the immunofluorescence assays, FFPE sections were cut into 4-μm thick sections. Xylene and ethanol were used for deparaffinization and rehydration. The sections were stained with anti-goat DPP 4 polyclonal antibody (R&D Systems, Minneapolis, MN, USA), mouse anti-mouse aquaporin-1 (AQP-1) antibody (Abcam, Cambridge, UK), and sera from MERS-CoV patients (patient's serum 9 weeks after symptom onset and the serum from a convalescent plasma donor; 1:2,000 dilution) as primary antibodies in a blocking reagent overnight at 4°C. Alex Fluor 555-conjugated anti-goat, Alex Fluor 647-conjugated antimouse (Molecular Probes, Eugene, OR, USA), and Alex Fluor 488-conjugated human IgG (Molecular Probes) were used as secondary antibodies. The sections were washed and incubated for an additional 5 minutes with 4´6-diamidino-2-phenylin- dole (DAPI) (Molecular Probes) for counterstaining. For the negative controls, the primary antibodies were omitted. We could not verify viral particles in the sections of in situ hybridization (Fig. 3A) . No evidence of MERS-CoV (human IgG) co-stained with DPP 4 was observed with confocal microscopy (Fig. 3B ).