Related Experiment Videos
Enterovirus isolation from children with acute respiratory infections and presumptive identification by a modified
Katsumi Mizuta1, Chieko Abiko, Hiroko Goto
1Department of Microbiology, Yamagata Prefectural Institute of Public Health, Tokamachi, Yamagata-shi, Japan. mizutak@pref.yamagata.jp
Objective:
To evaluate a modified microplate method, utilizing HEF, HEp-2, Vero, MDCK and newly introduced RD-18S and GMK cell lines, for virus isolation.
Methods:
From June to October 2001, 723 throat swab specimens taken from children with acute respiratory infections (ARIs) were inoculated onto these cells. To analyze cell sensitivity, we also inoculated 20 serotypes of stocked enteroviruses.
Results:
During the period, we isolated 40 Coxsackie A2 (CoxA2), 13 CoxA4, 16 CoxA16, 1 CoxB2, 11 CoxB3, 2 CoxB5, 54 echo16, 2 entero71 and 1 polio2. By observing a cell sensitivity pattern with HEF, HEp-2, Vero, RD-18S, and GMK, we could finally differentiate five enterovirus groups: CoxA except for CoxA16, CoxA16/entero71, CoxB, echovirus, and poliovirus.
Conclusions:
With this system, the RD-18S cell line enabled us to isolate CoxA virus, except for CoxA16, for the first time. Differentiation of five enterovirus groups by cell sensitivity simplified the specific identification by neutralization test as a presumptive identification. A modified microplate method may be an appropriate cell combination for virus isolation, especially for enteroviruses, and is expected to be used routinely for virologic diagnosis and to clarify the epidemiology of ARI in children.
Insights
A modified microplate method using RD-18S cell lines successfully isolated Coxsackie A virus, simplifying enterovirus identification. This method is suitable for routine virologic diagnosis and understanding acute respiratory infections (ARIs) in children.
Area of Science:
- Virology
- Microbiology
Background:
- Acute respiratory infections (ARIs) in children are often caused by enteroviruses.
- Accurate and efficient virus isolation is crucial for diagnosis and epidemiological studies.
Purpose of the Study:
- To evaluate a modified microplate method for virus isolation using multiple cell lines, including the novel RD-18S and GMK cell lines.
- To assess the sensitivity of different cell lines for isolating various enterovirus serotypes.
Main Methods:
- 723 throat swab specimens from children with ARIs were inoculated onto HEF, HEp-2, Vero, MDCK, RD-18S, and GMK cell lines.
- 20 enterovirus serotypes were inoculated to analyze cell sensitivity patterns.
Main Results:
- The study successfully isolated Coxsackie A2, Coxsackie A4, Coxsackie A16, Coxsackie B2, Coxsackie B3, Coxsackie B5, echovirus 16, enterovirus 71, and poliovirus 2.
- A cell sensitivity pattern allowed differentiation into five enterovirus groups: CoxA (except CoxA16), CoxA16/entero71, CoxB, echovirus, and poliovirus.
- The RD-18S cell line was effective for isolating CoxA viruses (excluding CoxA16).
Conclusions:
- The modified microplate method, particularly with the RD-18S cell line, offers a novel approach for enterovirus isolation.
- This method simplifies presumptive identification of enteroviruses, aiding in routine virologic diagnosis.
- The technique is expected to enhance the clarification of ARI epidemiology in pediatric populations.