Multi-site evaluation of the LN34 pan-lyssavirus real-time RT-PCR assay for post-mortem rabies diagnostics

• DOI: 10.1371/journal.pone.0197074
• Publication Date: 2018-05-16T17:42:59Z
• Authors: Crystal M. Gigante; Lisa Dettinger; James W. Powell; Melanie Seiders; Rene Edgar Condor...; Richard Griesser; Kenneth Okogi; Maria Carlos; Kendra Pesko; Mike Breckenridge; Edson Michael M. ...; Maria Yna Joyce V...; April D. Davis; Scott J. Brunt; Lillian Orciari; Pamela Yager; William C. Carson; Claire Hartloge; Jeremiah T. Saliki; Susan Sanchez; Mojgan Deldari; Kristina Hsieh; Ashutosh Wadhwa; Kimberly Wilkins; Veronica Yung Peredo; Patricia Rabideau; Nina Gruhn; Rolain Cadet; Shrikrishna Isloor; Sujith S. Nath; Tomy Joseph; Jinxin Gao; Ryan Wallace; Mary Reynolds; Victoria A. Olson; Yu Li
• Keywords: Lyssavirus; Gold standard (test); Direct fluorescent antibody; Multiplex

Rabies is a fatal zoonotic disease that requires fast, accurate diagnosis to prevent in an exposed individual. The current gold standard for post-mortem of human and animal rabies the direct fluorescent antibody (DFA) test. While DFA test has proven sensitive reliable, it high quality conjugates, skilled technician, fluorescence microscope diagnostic specimen sufficient quality. LN34 pan-lyssavirus real-time RT-PCR assay represents strong candidate diagnostics due its ability detect RNA across diverse Lyssavirus genus, sensitivity, potential use with deteriorated tissues, simple, easy implement design. Here, we present data from multi-site evaluation 14 laboratories. A total 2,978 samples (1,049 positive) Africa, Americas, Asia, Europe, Middle East were tested. exhibited low variability repeatability reproducibility studies was capable detecting viral fresh, frozen, archived, formalin-fixed brain tissue. displayed specificity (99.68%) sensitivity (99.90%) when compared test, no positive negative by assay. produced definitive findings 80 inconclusive or untestable DFA; 29 positive. Five assay, only one sample both tests. Furthermore, led identification false 11 results. Together, these results demonstrate reliability robustness support role improving surveillance.