tetano
Editor, Senior Moderator
Antimicrob Agents Chemother. 2013 Aug 5. [Epub ahead of print]
Bioluminescence-based neuraminidase inhibition assay for monitoring influenza drug susceptibility in clinical specimens.
Marjuki H, Mishin VP, Sleeman K, Okomo-Adhiambo M, Sheu TG, Guo L, Xu X, Gubareva LV.
Source
Virus Surveillance and Diagnosis Branch, Influenza Division, National Center for Immunization and Respiratory Diseases, Centers for Disease Control and Prevention, Atlanta, Georgia.
Abstract
The QFlu? prototype bioluminescence-based neuraminidase (NA) inhibition (NI) assay kit was designed to detect NA inhibitor (NAI)-resistant influenza viruses at point-of-care. Here we evaluated its suitability for drug susceptibility assessment at a surveillance laboratory. A comprehensive panel of reference viruses (n=14) and a set of 90 seasonal influenza A and B isolates were included for testing with oseltamivir and/or zanamivir in the QFlu assay using the manufacturer recommended protocol and a modified version to attune to surveillance requirement. The generated IC50s were compared with the NI assays currently used for monitoring influenza drug susceptibility, the fluorescent (FL) and chemiluminescent (CL) assays. To provide proof-of-principal, clinical specimens (n=235) confirmed by real-time RT-PCR to contain influenza A(H1N1)pdm09 virus and pre-screened for the oseltamivir resistance marker H275Y using pyrosequencing, were subsequently tested in the QFlu assay. All three NI assays were able to discriminate the reference NA variants and their matching wild-types based on the difference in their IC50s. Unless the antigenic types were first identified, certain NA variants (e.g. H3N2 with E119V) could only be detected among seasonal viruses using the FL assays. Notably, the QFlu assay identified oseltamivir-resistant A(H1N1)pdm09 viruses carrying the H275Y marker directly in clinical specimens, which is not feasible with the other two phenotypic assays that required prior virus culturing in cells. Furthermore, The QFlu assay allows detection of the influenza A and B virus isolates carrying established and potential NA inhibitor-resistance markers, and may become a useful tool for monitoring drug resistance in clinical specimens.
PMID:
23917311
[PubMed - as supplied by publisher]
http://www.ncbi.nlm.nih.gov/pubmed/23917311
Bioluminescence-based neuraminidase inhibition assay for monitoring influenza drug susceptibility in clinical specimens.
Marjuki H, Mishin VP, Sleeman K, Okomo-Adhiambo M, Sheu TG, Guo L, Xu X, Gubareva LV.
Source
Virus Surveillance and Diagnosis Branch, Influenza Division, National Center for Immunization and Respiratory Diseases, Centers for Disease Control and Prevention, Atlanta, Georgia.
Abstract
The QFlu? prototype bioluminescence-based neuraminidase (NA) inhibition (NI) assay kit was designed to detect NA inhibitor (NAI)-resistant influenza viruses at point-of-care. Here we evaluated its suitability for drug susceptibility assessment at a surveillance laboratory. A comprehensive panel of reference viruses (n=14) and a set of 90 seasonal influenza A and B isolates were included for testing with oseltamivir and/or zanamivir in the QFlu assay using the manufacturer recommended protocol and a modified version to attune to surveillance requirement. The generated IC50s were compared with the NI assays currently used for monitoring influenza drug susceptibility, the fluorescent (FL) and chemiluminescent (CL) assays. To provide proof-of-principal, clinical specimens (n=235) confirmed by real-time RT-PCR to contain influenza A(H1N1)pdm09 virus and pre-screened for the oseltamivir resistance marker H275Y using pyrosequencing, were subsequently tested in the QFlu assay. All three NI assays were able to discriminate the reference NA variants and their matching wild-types based on the difference in their IC50s. Unless the antigenic types were first identified, certain NA variants (e.g. H3N2 with E119V) could only be detected among seasonal viruses using the FL assays. Notably, the QFlu assay identified oseltamivir-resistant A(H1N1)pdm09 viruses carrying the H275Y marker directly in clinical specimens, which is not feasible with the other two phenotypic assays that required prior virus culturing in cells. Furthermore, The QFlu assay allows detection of the influenza A and B virus isolates carrying established and potential NA inhibitor-resistance markers, and may become a useful tool for monitoring drug resistance in clinical specimens.
PMID:
23917311
[PubMed - as supplied by publisher]
http://www.ncbi.nlm.nih.gov/pubmed/23917311