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Bioluminescence-based neuraminidase inhibition assay for monitoring influenza virus drug susceptibility in clinical specimens.
Antimicrob Agents Chemother 2013; 57(11):5209-15AA

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 virus 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 attuned to surveillance requirements. The 50% inhibitory concentrations (IC50s) generated were compared with those of NI assays currently used for monitoring influenza drug susceptibility, the fluorescent (FL) and chemiluminescent (CL) assays. To provide proof of principle, clinical specimens (n = 235) confirmed by real-time reverse transcription (RT)-PCR to contain influenza virus A(H1N1)pdm09 and prescreened 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-type viruses based on the difference in their IC50s. Unless the antigenic types were first identified, certain NA variants (e.g., H3N2 with E119V) could be detected among seasonal viruses using the FL assays only. 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, which required prior virus culturing in cells. Furthermore, The QFlu assay allows detection of the influenza virus A and B isolates carrying established and potential NA inhibitor resistance markers and may become a useful tool for monitoring drug resistance in clinical specimens.

Authors+Show Affiliations

Virus Surveillance and Diagnosis Branch, Influenza Division, National Center for Immunization and Respiratory Diseases, Centers for Disease Control and Prevention, Atlanta, Georgia, USA.No affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info available

Pub Type(s)

Journal Article

Language

eng

PubMed ID

23917311

Citation

Marjuki, Henju, et al. "Bioluminescence-based Neuraminidase Inhibition Assay for Monitoring Influenza Virus Drug Susceptibility in Clinical Specimens." Antimicrobial Agents and Chemotherapy, vol. 57, no. 11, 2013, pp. 5209-15.
Marjuki H, Mishin VP, Sleeman K, et al. Bioluminescence-based neuraminidase inhibition assay for monitoring influenza virus drug susceptibility in clinical specimens. Antimicrob Agents Chemother. 2013;57(11):5209-15.
Marjuki, H., Mishin, V. P., Sleeman, K., Okomo-Adhiambo, M., Sheu, T. G., Guo, L., ... Gubareva, L. V. (2013). Bioluminescence-based neuraminidase inhibition assay for monitoring influenza virus drug susceptibility in clinical specimens. Antimicrobial Agents and Chemotherapy, 57(11), pp. 5209-15. doi:10.1128/AAC.01086-13.
Marjuki H, et al. Bioluminescence-based Neuraminidase Inhibition Assay for Monitoring Influenza Virus Drug Susceptibility in Clinical Specimens. Antimicrob Agents Chemother. 2013;57(11):5209-15. PubMed PMID: 23917311.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Bioluminescence-based neuraminidase inhibition assay for monitoring influenza virus drug susceptibility in clinical specimens. AU - Marjuki,Henju, AU - Mishin,Vasiliy P, AU - Sleeman,Katrina, AU - Okomo-Adhiambo,Margaret, AU - Sheu,Tiffany G, AU - Guo,Lizheng, AU - Xu,Xiyan, AU - Gubareva,Larisa V, Y1 - 2013/08/05/ PY - 2013/8/7/entrez PY - 2013/8/7/pubmed PY - 2014/5/16/medline SP - 5209 EP - 15 JF - Antimicrobial agents and chemotherapy JO - Antimicrob. Agents Chemother. VL - 57 IS - 11 N2 - 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 virus 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 attuned to surveillance requirements. The 50% inhibitory concentrations (IC50s) generated were compared with those of NI assays currently used for monitoring influenza drug susceptibility, the fluorescent (FL) and chemiluminescent (CL) assays. To provide proof of principle, clinical specimens (n = 235) confirmed by real-time reverse transcription (RT)-PCR to contain influenza virus A(H1N1)pdm09 and prescreened 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-type viruses based on the difference in their IC50s. Unless the antigenic types were first identified, certain NA variants (e.g., H3N2 with E119V) could be detected among seasonal viruses using the FL assays only. 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, which required prior virus culturing in cells. Furthermore, The QFlu assay allows detection of the influenza virus A and B isolates carrying established and potential NA inhibitor resistance markers and may become a useful tool for monitoring drug resistance in clinical specimens. SN - 1098-6596 UR - https://www.unboundmedicine.com/medline/citation/23917311/Bioluminescence_based_neuraminidase_inhibition_assay_for_monitoring_influenza_virus_drug_susceptibility_in_clinical_specimens_ L2 - http://aac.asm.org/cgi/pmidlookup?view=long&pmid=23917311 DB - PRIME DP - Unbound Medicine ER -