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Comparative sensitivity of various cell culture systems for isolation of viruses from wastewater and fecal samples
Abstract:
In efforts to define the most sensitive cell culture systems for recovery of viruses from wastewaters, 181 samples were inoculated in parallel into tube cultures of various cell types and were plaqued in bottle and petri dish cultures of three types of monkey kidney cells. Polioviruses were recovered most frequently in the RD line of human rhabdomyosarcoma cells, group A coxsackieviruses in RD and human fetal diploid kidney (HFDK) cells, group B coxsackieviruses in the BGM line of African green monkey kidney cells, echoviruses in RD and primary rhesus monkey kidney (RhMK) cells, and reoviruses in RhMK cells. BGM cells were unsatisfactory for recovery of viruses other than polioviruses and group B coxsackieviruses, and a line of fetal rhesus monkey kidney (MFK) was not a satisfactory substitute for primary RhMK. With RhMK cells, comparable numbers of virus isolations were made in tube cultures and in plaque assays conducted in bottle cultures, but with BGM and MFK cells, fewer isolations were made by plaquing than by inoculation of tube cultures. In comparative plaque assays on fecal samples under three different overlays in bottle and plate cultures of RhMK, BGM, and MFK cells, it was found that plaquing in the most sensitive system, RhMK, was less efficient for virus recovery than was inoculation of tube cultures of RhMK or HFDK cells. Overall, plaque assays performed in petri dishes in a CO(2) incubator yielded fewer virus isolates than did parallel plaque assays performed in closed bottle cultures. Other limitations of plaque assays for recovery of human enteric viruses are discussed.
Insights
RD and human fetal diploid kidney (HFDK) cells are most sensitive for recovering enteric viruses from wastewater. Plaque assays in petri dishes were less efficient than tube cultures for virus recovery.
Area of Science:
- Virology
- Environmental Microbiology
- Cell Culture Technology
Background:
- Wastewater surveillance is crucial for monitoring enteric virus circulation.
- Identifying optimal cell culture systems is essential for sensitive virus detection.
Purpose of the Study:
- To evaluate and compare the efficiency of various cell culture systems for recovering human enteric viruses from wastewater samples.
- To determine the most sensitive cell lines and methods for virus isolation.
Main Methods:
- 181 wastewater samples were inoculated into diverse cell cultures.
- Virus recovery was assessed using tube cultures and plaque assays in bottle and petri dish cultures.
- Different cell types, including RD, HFDK, BGM, RhMK, and MFK, were utilized.
Main Results:
- RD cells showed highest recovery for polioviruses and group A coxsackieviruses.
- HFDK cells were also effective for group A coxsackieviruses.
- BGM cells were best for group B coxsackieviruses, while RhMK cells were optimal for reoviruses.
- Plaque assays, especially in petri dishes, were less efficient than tube cultures for virus isolation.
Conclusions:
- RD and HFDK cell lines are recommended for sensitive recovery of various enteric viruses from wastewater.
- Tube cultures generally outperform plaque assays for virus isolation efficiency.
- Cell culture system selection significantly impacts virus recovery rates in wastewater surveillance.