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[Application of a diagnostic method using reverse transcription-PCR ELISA for the diagnosis of enteroviral
Kwisung Park1, Kangbum Lee, Kyungah Baek
1Chungcheongnam-do Health & Environment Research Institute, Daejeon, Korea.
Background:
Enteroviruses are known as major pathogen for aseptic meningitis. Although rapid diagnosis for enteroviruses is very essential to exclude bacterial infections in patients with meningitis, classical diagnostic method based on virus isolation is not practicable for timely treatment of patients due to its laborious and time-consuming procedure. Recently molecular methodologies as alternatives are routinely used for rapid and sensitive diagnosis for enteroviruses infections.
Methods:
Reverse transcription (RT)-PCR ELISA kit for targeting 5' non-coding region (NCR) with highly conserved genetic identity among all genotypes of enteroviruses was introduced in this investigation. RT-PCR ELISA was evaluated about sensitivity and specificity through virus isolation using clinical specimens from patients suspected of enteroviral infections and enteroviral isolates comparing with conventional RT-PCR identifying them.
Results:
The detection limit of the RT-PCR ELISA was up to 10-100 folds higher than virus isolation using cell culture and conventional RT-PCR. On comparison between above two methods, the detection rate of RT-PCR ELISA for clinical specimens from patients with aseptic meningitis was 7% higher than that of conventional RT-PCR targeting 5'NCR (P=0.016).
Conclusions:
Our results suggest that RT-PCR ELISA developed in this study could be an alternative diagnostic method for the detection of enteroviral genome with high sensitivity and specificity.
Insights
A new RT-PCR ELISA test offers a highly sensitive and specific method for detecting enteroviruses, crucial pathogens in aseptic meningitis. This molecular technique provides a faster alternative to traditional virus isolation for timely patient treatment.
Area of Science:
- Virology
- Molecular Biology
- Infectious Diseases
Context:
- Enteroviruses are a primary cause of aseptic meningitis.
- Traditional virus isolation is slow, hindering timely treatment.
- Molecular methods offer rapid and sensitive diagnostics for enterovirus infections.
Purpose:
- To evaluate a novel Reverse Transcription Polymerase Chain Reaction (RT-PCR) ELISA kit for enterovirus detection.
- To assess the sensitivity and specificity of the RT-PCR ELISA compared to conventional methods.
- To determine the utility of RT-PCR ELISA for diagnosing enteroviral meningitis.
Summary:
- A new RT-PCR ELISA kit targeting the conserved 5' non-coding region (NCR) of enteroviruses was developed and evaluated.
- The RT-PCR ELISA demonstrated a 10-100 fold higher detection limit than virus isolation and conventional RT-PCR.
- This method showed a 7% higher detection rate in clinical meningitis specimens compared to conventional RT-PCR (P=0.016).
Impact:
- The developed RT-PCR ELISA serves as a viable alternative diagnostic tool for enteroviral infections.
- This assay offers improved sensitivity and specificity for detecting the enteroviral genome.
- Faster diagnosis can lead to quicker treatment decisions for patients with suspected meningitis.
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