BACKGROUND: Australian bat lyssavirus (ABLV) has been transmitted to humans following a scratch or bite from an infected bat in two cases. Following a scratch or bite to a person, the bat is usually submitted for testing and diagnosis is made using a direct fluorescent antibody test on a brain smear. A nested RT-PCR assay has also been utilised to confirm diagnosis. If positive for lyssavirus, post-exposure prophylaxis is administered. OBJECTIVES: The TaqMan assay was developed to improve the diagnosis of ABLV infection, following problems encountered with the generation of spurious PCR products in the nested RT-PCR and also to reduce the high risk of contamination inherent with nested PCRs. STUDY DESIGN: RNA was extracted from 161 bat brains and the samples were compared using a conventional RT-PCR and the TaqMan based assay. Samples from a patient with an ABLV infection collected antemortem and postmortem were also tested. RESULTS: The sensitivity of the new TaqMan based PCR assay compared favourably with the nested PCR previously in use in our laboratory. This assay was able to detect RNA in samples collected antemortem and postmortem for the diagnosis of a human case of ABLV. CONCLUSIONS: The major advantage of the TaqMan based assay was the speed of diagnosis with a result within minutes of completing the PCR (a result within 4 h of receiving the specimen). This test greatly reduces the chance of false positives through the elimination of second-round PCR and the requirement for agarose gels. The assay is sensitive and specific and should be invaluable for future antemortem and postmortem diagnosis of ABLV infection in humans.
BACKGROUND:Australian bat lyssavirus (ABLV) has been transmitted to humans following a scratch or bite from an infected bat in two cases. Following a scratch or bite to a person, the bat is usually submitted for testing and diagnosis is made using a direct fluorescent antibody test on a brain smear. A nested RT-PCR assay has also been utilised to confirm diagnosis. If positive for lyssavirus, post-exposure prophylaxis is administered. OBJECTIVES: The TaqMan assay was developed to improve the diagnosis of ABLV infection, following problems encountered with the generation of spurious PCR products in the nested RT-PCR and also to reduce the high risk of contamination inherent with nested PCRs. STUDY DESIGN: RNA was extracted from 161 bat brains and the samples were compared using a conventional RT-PCR and the TaqMan based assay. Samples from a patient with an ABLV infection collected antemortem and postmortem were also tested. RESULTS: The sensitivity of the new TaqMan based PCR assay compared favourably with the nested PCR previously in use in our laboratory. This assay was able to detect RNA in samples collected antemortem and postmortem for the diagnosis of a human case of ABLV. CONCLUSIONS: The major advantage of the TaqMan based assay was the speed of diagnosis with a result within minutes of completing the PCR (a result within 4 h of receiving the specimen). This test greatly reduces the chance of false positives through the elimination of second-round PCR and the requirement for agarose gels. The assay is sensitive and specific and should be invaluable for future antemortem and postmortem diagnosis of ABLV infection in humans.
Authors: David Warrilow; Bruce Harrower; Ina L Smith; Hume Field; Roscoe Taylor; Craig Walker; Greg A Smith Journal: Emerg Infect Dis Date: 2003-02 Impact factor: 6.883
Authors: Peter H Holz; Linda F Lumsden; Julian Druce; Alistair R Legione; Paola Vaz; Joanne M Devlin; Jasmin Hufschmid Journal: PLoS One Date: 2018-05-24 Impact factor: 3.240
Authors: Crystal M Gigante; Lisa Dettinger; James W Powell; Melanie Seiders; Rene Edgar Condori Condori; Richard Griesser; Kenneth Okogi; Maria Carlos; Kendra Pesko; Mike Breckenridge; Edson Michael M Simon; Maria Yna Joyce V Chu; 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 Journal: PLoS One Date: 2018-05-16 Impact factor: 3.240