title: Comparison of Automated Quantitative Reverse Transcription-PCR and Direct Fluorescent-Antibody Detection for Routine Rabies Diagnosis in the United States.
abstract: Rabies virus found worldwide and prevalent throughout the United States continues to be a public health concern. Direct-fluorescent antibody (DFA) detection remains the gold standard for rabies virus diagnostics. Assessing the utility of a high-throughput molecular platform such as the QIAsymphony SP/AS, in conjunction with quantitative reverse transcription-PCR (qRT-PCR), to augment or potentially replace the DFA test, was the focus of this project. Here we describe a triplex qRT-PCR assay, including assembly and evaluation for sensitivity, specificity, and ability to detect variants. Additionally, we compared the qRT-PCR assay to the gold standard direct fluorescent-antibody test. More than 1,000 specimens submitted for routine rabies diagnosis were tested to directly compare the two methods. All results were in agreement between the two methods, with one additional specimen detected by qRT-PCR below the limits of the DFA sensitivity. With the proper continued validation for variant detection, molecular methods have a place in routine rabies diagnostics within the United States.
authors.affiliation: Wadsworth Center, New York State Department of Health, Albany, New York, USA.
authors.forename: Michelle
authors.initials: M
authors.lastname: Dupuis
authors.affiliation: Wadsworth Center, New York State Department of Health, Albany, New York, USA.
authors.forename: Scott
authors.initials: S
authors.lastname: Brunt
authors.affiliation: Wadsworth Center, New York State Department of Health, Albany, New York, USA.
authors.forename: Kim
authors.initials: K
authors.lastname: Appler
authors.affiliation: Wadsworth Center, New York State Department of Health, Albany, New York, USA april.davis@health.ny.gov.
authors.forename: April
authors.initials: A
authors.lastname: Davis
authors.affiliation: Wadsworth Center, New York State Department of Health, Albany, New York, USA.
authors.forename: Robert
authors.initials: R
authors.lastname: Rudd
mesh_terms: D000818:Animals; D000914:Antibodies, Viral / Q000097:blood*; D001921:Brain / Q000821:virology; D019085:Fluorescent Antibody Technique, Direct / Q000379:methods*; D006801:Humans; D060885:Multiplex Polymerase Chain Reaction / Q000379:methods; D012367:RNA, Viral / Q000032:analysis*; D011818:Rabies / Q000175:diagnosis*; D060888:Real-Time Polymerase Chain Reaction / Q000379:methods*; D012680:Sensitivity and Specificity; D014481:United States
chemical_list: D000914:Antibodies, Viral; D012367:RNA, Viral