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      Assays for laboratory confirmation of novel human coronavirus (hCoV-EMC) infections.

      Euro surveillance : bulletin Européen sur les maladies transmissibles = European communicable disease bulletin

      Coronavirus, classification, genetics, isolation & purification, Coronavirus Infections, diagnosis, virology, Fluorescent Antibody Technique, Germany, Humans, Laboratories, standards, Polymorphism, Restriction Fragment Length, RNA, Viral, blood, Reverse Transcriptase Polymerase Chain Reaction, methods, Sensitivity and Specificity, Sequence Analysis, DNA, Virology

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          Abstract

          We present a rigorously validated and highly sensitive confirmatory real-time RT-PCR assay (1A assay) that can be used in combination with the previously reported upE assay. Two additional RT-PCR assays for sequencing are described, targeting the RdRp gene (RdRpSeq assay) and N gene (NSeq assay), where an insertion/deletion polymorphism might exist among different hCoV-EMC strains. Finally, a simplified and biologically safe protocol for detection of antibody response by immunofluorescence microscopy was developed using convalescent patient serum.

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          23231891

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