Rapid culture for influenza virus, types A and B, in 96-well plates

Laboratory-based assessment of influenza in German ambulant patients from 1998 to 2008

“…Influenza A and B epidemics usually began in January in calendar weeks 1-3 and sometimes later in February in calendar weeks 5-6 (1997/1998, 2001/2002, and 2005/2006 seasons). Epidemics reached the highest level in February through the first half of March (calendar weeks 5-10) and ended in the second half of March through April (calendar weeks [12][13][14][15][16]. These data are in agreement with the data for the whole of Germany, published every year by the Robert Koch Institute [23].…”

“…The speed of the test is realized by the centrifugation of vials or plates, which enhances cell infection [13], and by the precytopathic detection of infected cells using staining with peroxidase-or fluorescein isothiocyanate-labeled MAbs. The sensitivity of rapid precytopathic detection of influenza 1 or 2 days after inoculation has in some studies been reported to be higher than that of CCC [14,15], but other authors have found a similar sensitivity for both techniques [16,17] or a higher sensitivity for CCC [18,19]. In our study, the sensitivity of CCC was higher, equal, or lower than that of RCA in the different influenza seasons.…”

Simultaneous rapid culture for four respiratory viruses in the same cell monolayer using a differential multicolored fluorescent confirmatory stain

“…The reference methods used for the four viruses are listed in Table 1. The rapid culture for influenza virus types A and B was performed in 96-well plates as described previously (1). Specimens with discrepant results were cultured by conventional tube culture with hemadsorption (3).…”

“…Conventional virus culture, however, can require up to 2 weeks for a final result; this is too slow to be useful in patient treatment and management. Many laboratories currently employ rapid-culture methods, with sensitivities approaching or even exceeding those of conventional culture for isolation of respiratory viruses (1,7). Results are usually available within 48 h, but separate tests must be run for each suspected virus.…”

“…Simultaneous rapid culture for adenovirus, cytomegalovirus, and herpes simplex virus in the same shell vial with a multicolored differential fluorescent confirmatory stain has been reported (2). We used the same stain (three fluorochromes) to differentiate respiratory viruses isolated in a 96-well plate format that was previously described for influenza virus culture (1). We expanded the culture-confirmatory stain by adding a fourth fluorescent marker and used it to detect and differentiate among four viruses.…”