Comparing molecular methods for early detection and serotyping of enteroviruses in throat swabs of pediatric patients
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Published on: July 22, 2012
Pai-Shan Chiang1, Mei-Liang Huang, Shu-Ting Luo
1National Institutes of Infectious Diseases and Vaccinology, National Health Research Institutes, Taiwan.
Background:
Enteroviruses include over 100 serotypes and usually cause self-limited infections with non-specific symptoms in children, with the exceptions of polioviruses and enterovirus 71 which frequently cause neurologic complications. Therefore, early detection and serotyping of enteroviruses are critical in clinical management and disease surveillance. Traditional methods for detection and serotyping of enteroviruses are virus isolation and immunofluorescence assay, which are time-consuming. In this study, we compare virus isolation and two molecular tests for detection and serotyping of enteroviruses in clinical samples.
Methods:
One hundred and ten throat swabs were collected from pediatric outpatients with enterovirus-like illnesses (hand-foot-mouth disease, herpangina, and non-specific febrile illness). Virus isolation was conducted using multiple cell lines and isolated viruses were serotyped using immunofluorescent assay. In the molecular tests, a semi-nested RT-PCR and a novel CODEHOP platform were used to detect the 5'UTR and VP1 genes of enteroviruses, respectively. Amplified nucleotides were sequenced and genotyped.
Results:
Among the 110 cases, 39(35%), 52(47%), and 46(42%) were tested positive with these three tests, respectively. Using the consensus results of these three tests as the gold standard, agreement of the VP1 CODEHOP test was 96%, which is higher than those of the virus isolation (89%) and the 5'-UTR test (88%). The VP1 CODEHOP test also has the best performance on serotyping confirmed with serum neutralization tests.
Conclusions:
The VP1 CODEHOP test performed well for detection and serotyping of enteroviruses in clinical specimens and could reduce unnecessary hospitalization cares during enterovirus seasons.
Insights
The VP1 CODEHOP molecular test accurately detects and serotypes enteroviruses in children, offering a faster alternative to traditional methods. This advancement aids clinical management and disease surveillance for enterovirus infections.
Area of Science:
- Virology
- Molecular Diagnostics
- Pediatric Infectious Diseases
Background:
- Enteroviruses encompass over 100 serotypes, typically causing mild childhood illnesses, but some strains like poliovirus and enterovirus 71 can lead to severe neurological complications.
- Prompt identification and serotyping of enteroviruses are crucial for effective clinical care and public health surveillance.
- Conventional methods such as virus isolation and immunofluorescence assay are time-consuming for enterovirus detection and serotyping.
Purpose of the Study:
- To compare the diagnostic performance of virus isolation with two molecular tests for enterovirus detection and serotyping.
- To evaluate the efficacy of a novel CODEHOP platform targeting the VP1 gene against a semi-nested RT-PCR targeting the 5'UTR region.
Main Methods:
- Throat swabs from 110 pediatric outpatients with enterovirus-like illnesses were analyzed.
- Virus isolation was performed using multiple cell lines, followed by immunofluorescence assay for serotyping.
- Molecular detection involved a semi-nested RT-PCR for the 5'UTR and a CODEHOP platform for the VP1 gene, with subsequent sequencing and genotyping.
Main Results:
- The VP1 CODEHOP test demonstrated the highest agreement (96%) with the consensus results, surpassing virus isolation (89%) and the 5'-UTR test (88%).
- The VP1 CODEHOP test also showed superior performance in serotyping, confirmed by serum neutralization tests.
- Positive detection rates were 35% for virus isolation, 47% for the 5'-UTR test, and 42% for the VP1 CODEHOP test.
Conclusions:
- The VP1 CODEHOP test is a reliable method for detecting and serotyping enteroviruses in clinical samples.
- This molecular approach can potentially decrease unnecessary hospitalizations during peak enterovirus seasons.
