Rapid and sensitive detection of respiratory virus infections for directed antiviral treatment using R-Mix cultures.
BACKGROUND
The development of new anti-influenza drugs has led to concerns regarding the impact on healthcare costs if they are used indiscriminately. Restricting their use to proven influenza virus infections has the potential to overcome costly inappropriate therapy. However, conventional culture (CC) does not generate results quickly enough to facilitate the timely initiation of treatment, and rapid detection tests have suboptimal sensitivity. We therefore investigated a new rapid culture system (R-Mix) that contains a mixture of two cell lines and detects respiratory viruses within 24 h.


OBJECTIVE
To compare the analytical sensitivity of R-Mix with CC and rapid detection methods, for the detection of influenza and other respiratory viruses. To compare the clinical sensitivity of R-Mix with CC and direct antigen detection for the detection of respiratory viruses in primary and acute care settings.


METHODS
Stock cultures of influenza virus were titrated and tested by R-Mix, ZstatFlu and FLU OIA. Stock cultures of adenovirus and parainfluenza virus type 3 were titrated and tested by R-Mix and CC. Specimens, which had previously tested positive for influenza viruses, were titrated and tested by R-Mix and CC. In symptomatic patients, the majority of whom were from primary care settings, 124 sequential specimens were tested for influenza viruses by immunofluorescent direct antigen detection and R-Mix. A separate set of 111 sequential specimens, from various symptomatic patient groups, were tested for influenza viruses by CC and R-Mix. Additionally, in acute care patients being surveillance tested during periods of immunosuppression, 155 specimens were tested for respiratory viruses (influenza A and B, parainfluenza 1-3, adenovirus and respiratory syncytial virus (RSV)) by CC and R-Mix.


RESULTS
With titrated stock cultures, R-Mix showed an analytical limit of detection of ten infectious virus particles per vial for influenza A, compared with 100,000 particles per test for FLU OIA and 1,000,000 for ZstatFlu. R-Mix also showed a 100-fold greater sensitivity for the detection of influenza A and equivalent sensitivity for the detection of influenza B when compared with CC in titrated known positive specimens. Further, it showed equivalent sensitivity to CC for the detection of adenovirus and parainfluenza virus type 3 in titrated stock cultures. Among prospective specimens from symptomatic patients, the sensitivity of R-Mix, CC and direct antigen detection tests (DAT) for influenza virus detection, was 100, 67 and 66%, respectively, and the specificity was 100, 100 and 98%, respectively. In surveillance specimens from immunosuppressed patients, the sensitivities of R-Mix and CC for respiratory virus detection were equivalent. Moreover, R-Mix results were available within 24 h, and by altering the antibody staining reagents either influenza viruses, or all seven major respiratory viruses, could be detected and distinguished in a single test.


CONCLUSIONS
R-Mix is a simple, rapid and sensitive system for the detection of influenza viruses that facilitates the restriction of antiviral drugs to patients with culture-confirmed infections.
St  George
Kirsten
K
Clinical Virology Laboratory, University of Pittsburgh Medical Center, A-912, Presbyterian, 200 Lothrop Street, Pittsburgh, PA 15213, USA. stgeorgek@msx.upmc.edu
Patel
Navin M
NM
Hartwig
Robert A
RA
Scholl
David R
DR
Jollick
Joseph A
JA
Kauffmann
Lynn M
LM
Evans
Martin R
MR
Rinaldo
Charles R
CR
D000256:Adenoviridae / Q000302:isolation & purification; D002460:Cell Line; D019085:Fluorescent Antibody Technique, Direct; D006801:Humans; D007251:Influenza, Human / Q000175:diagnosis / Q000821:virology; D009975:Orthomyxoviridae / Q000302:isolation & purification; D010224:Parainfluenza Virus 3, Human / Q000302:isolation & purification; D011933:Reagent Kits, Diagnostic; D012141:Respiratory Tract Infections / Q000175:diagnosis* / Q000821:virology; D010253:Respirovirus Infections / Q000175:diagnosis / Q000821:virology; D013997:Time Factors
D011933:Reagent Kits, Diagnostic