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Abstract
Background
Enteroviruses are the most frequent etiologic agents of aseptic meningitis and are estimated to be the cause of 70% to 90% of viral meningitis cases. Enterovirus diagnosis can be difficult because clinical features vary according to patient immunity and age. The purpose of this study was to evaluate the performance of the realtime nucleic acid sequence-based amplification (NASBA) assay compared to that of the realtime nested RT-PCR assay for enterovirus detection.
Methods
This study was performed on 96 patients suspected of aseptic meningitis based on clinical features. RNA was extracted using NucliSENS EasyMAG and realtime NASBA assay was performed using NucliSENS EasyQ Enterovirus and NucliSENS EasyQ Basic 2. We also executed in-house realtime nested RT-PCR assay for RNA extracted via QIAamp Viral RNA Mini.
Results
The positive rate of realtime NASBA assay was 45.8% for enterovirus detection. The positive rate of first realtime reverse transcription PCR was 22.9% and the second realtime PCR was 57.3%. The concordant rate of the realtime NASBA assay and first realtime reverse transcription PCR was 75.0%. The concordant rate of the realtime NASBA assay and second realtime PCR was 86.5%.
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Fig. 1.
Detection of enterovirus by nucleic acid sequence-based amplification and realtime nested reverse transcription PCR. (A) Nucleic acid sequence-based amplification. ①∼ ② and ④∼⑥, samples with a internal control signal (red) and no enterovirus signal (blue); ③ and ⑦, samples with a internal control signal and a strong enterovirus signal. (B) The first realtime reverse transcription PCR. ③, positive for enterovirus. (C) The nested realtime PCR. ③ and ⑦, positive for enterovirus; PC, positive control.
Table 1.
Comparison of results of enterovirus detection by realtime nucleic acid sequence-based amplification between NucliSENS EasyQ Basic 1 and NucliSENS EasyQ Basic 2
| NucliSENS EasyQ Basic 2 | NucliSENS EasyQ Basic 1 | Total | ||
|---|---|---|---|---|
| Positive | Negative | Invalid | ||
| Positive | 5 | 2 | 7 | |
| Negative | 67 | 8 | 75 | |
| Invalid | 1 | 1 | 2 | |
| Total | 5 | 70 | 9 | 84 |
Table 2.
Comparison of results between realtime nested reverse transcription PCR and realtime nucleic acid sequence-based amplification for detection of enterovirus
| NASBA | First realtime RT-PCR | Nested realtime PCR | Total | ||
|---|---|---|---|---|---|
| Positive | Negative | Positive | Negative | ||
| Positive | 21 | 23 | 43 | 1 | 44 |
| Negative | 1 | 51 | 12 | 40 | 52 |
| Total | 22 | 74 | 55 | 41 | 96 |
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