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A microplate method for isolation of viruses from infants and children with acute respiratory infections
1Virus Center, Sendai National Hospital, Miyagi.
Abstract:
Between December 1984 and December 1986, a microplate technique was adopted for isolation of viruses from infants and children with acute respiratory infections. By using two kinds of tissue culture microplates, i.e., the HHVM plate, containing human embryonic fibroblast (HEF), HEp-2, Vero and MDCK cells, and the MK plate which contains secondary monkey kidney cells, 1,080 field viruses were isolated from 1,061 (24.9%) out of 4,254 throat swabs. Of these 1,080 isolates, 1,003 (92.9%) were recovered in the HHVM plates and the remaining 77 (7.1%) in the MK plates. With the HHVM plate, influenza A and B viruses were cultivated in MDCK, RS virus in HEp-2, parainfluenza and mumps viruses in Vero, adenoviruses in both HEF and HEp-2, polioviruses in HEF, HEp-2 and Vero, coxsackie B viruses in both HEp-2 and Vero, rhino and echo viruses in HEF, herpes simplex virus in both HEF and HEp-2, and cytomegalovirus in HEF, although MK were more sensitive than Vero to parainfluenza and coxsackie B viruses. There was no difference in the rate of isolation of viruses between the microplate and ordinary tube methods. Cross contamination in the microplates was negligible for routine work.
Insights
A microplate technique effectively isolated viruses from children with respiratory infections. This method showed comparable isolation rates to traditional methods, with minimal cross-contamination, proving its utility in virology.
Area of Science:
- Virology
- Microbiology
- Infectious Diseases
Background:
- Acute respiratory infections (ARIs) in infants and children pose a significant public health challenge.
- Accurate and efficient viral isolation is crucial for diagnosis and understanding ARI epidemiology.
Purpose of the Study:
- To evaluate the efficacy of a microplate technique for isolating viruses from pediatric patients with ARIs.
- To compare the performance of two types of microplates (HHVM and MK) for viral isolation.
Main Methods:
- A microplate technique using HHVM and MK plates was employed for viral isolation from 4,254 throat swabs collected between December 1984 and December 1986.
- Specific cell lines (human embryonic fibroblast, HEp-2, Vero, MDCK, and monkey kidney cells) were utilized within the microplates.
- Isolated viruses were identified based on their growth characteristics in different cell lines.
Main Results:
- A total of 1,080 viruses were isolated from 1,061 patients (24.9% isolation rate).
- The HHVM plate yielded 92.9% of the isolates, while the MK plate yielded 7.1%.
- The microplate technique demonstrated comparable viral isolation rates to conventional tube methods, with negligible cross-contamination.
Conclusions:
- The microplate technique is a viable and efficient method for isolating a broad spectrum of viruses causing ARIs in children.
- The HHVM plate proved more effective for isolating common respiratory viruses compared to the MK plate.
- This method offers a reliable alternative to traditional techniques for routine viral diagnostics.