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A one-step RT-PCR assay using an enzyme-linked detection system for the diagnosis of enterovirus meningitis
K A Stellrecht1, I Harding, F M Hussain
1Department of Pathology and Laboratory Medicine, Albany Medical College, 47 New Scotland Ave., Albany, NY 12208, USA.
Background:
Enteroviruses are the most common cause of meningitis in the United States, with an estimated 50000-75000 cases each year. Enteroviral meningitis (EVM) is frequently a diagnosis of exclusion, as viral cultures lack sensitivity and require prolonged incubation periods.
Objective:
To develop a sensitive and rapid test for the diagnosis of EVM.
Study Design:
A rapid, one-step, reverse transcriptase-polymerase chain reaction (RT-PCR) was used in a prospective analysis of 160 patients who had cerebrospinal fluid (CSF) tested for enterovirus.
Results:
Of the 160 patients, 14 were excluded due to missing CSF viral culture data. A total of 14 were CSF culture positive (10 with pleocytosis) and 19 were PCR positive (15 with pleocytosis). The ability to detect enterovirus by either culture or PCR correlated significantly with the white blood cell count in the CSF (P=0.001). Based on a clinical definition of enterovirus culture positive and pleocylosis: ten had definite EVM and 12 had probable EVM (pleocytosis without any other cause). Four had possible EVM (CSF culture positive without pleocytosis) and 18 had pleocytosis due to other causes. PCR was positive in all ten patients with definite EVM. Five out of 12 patients with probable EVM and three out of four patients with possible EVM. No patients with pleocytosis due to other causes were PCR positive and one patient that was defined as EVM negative (culture negative and no pleocytosis) was PCR positive. Overall, PCR was positive in 18 out of the 26 patients with a likelihood of EVM, while CSF culture was positive in only 14 cases. Our results demonstrated that RT-PCR enhances the sensitivity of enterovirus detection in CSF (69 vs. 54% for culture).
Conclusion:
The diagnosis of EVM is difficult to make clinically: the enhanced sensitivity and rapid turn around time of PCR will be of great clinical benefit.
Insights
A rapid reverse transcriptase-polymerase chain reaction (RT-PCR) test offers a more sensitive and faster method for diagnosing enteroviral meningitis (EVM). This molecular diagnostic approach significantly improves upon traditional viral cultures for detecting enteroviruses in cerebrospinal fluid.
Area of Science:
- Virology
- Infectious Diseases
- Molecular Diagnostics
Background:
- Enteroviruses are a leading cause of meningitis in the United States, resulting in approximately 50,000-75,000 cases annually.
- Diagnosing enteroviral meningitis (EVM) is often challenging due to the low sensitivity and lengthy incubation periods associated with conventional viral cultures.
- EVM is frequently a diagnosis of exclusion, necessitating improved diagnostic tools.
Purpose of the Study:
- To develop a highly sensitive and rapid diagnostic test for enteroviral meningitis (EVM).
- To evaluate the efficacy of a novel molecular diagnostic method for enterovirus detection in cerebrospinal fluid (CSF).
Main Methods:
- A prospective analysis involving 160 patients with CSF samples tested for enterovirus.
- Utilized a rapid, one-step, reverse transcriptase-polymerase chain reaction (RT-PCR) assay.
- Compared RT-PCR results with traditional CSF viral culture and clinical findings, including pleocytosis.
Main Results:
- RT-PCR demonstrated enhanced sensitivity in detecting enterovirus in CSF, with a positivity rate of 69% compared to 54% for CSF culture.
- The study identified 18 patients with a likelihood of EVM using RT-PCR, versus 14 using CSF culture.
- RT-PCR detected enterovirus in all definite EVM cases and a significant proportion of probable and possible EVM cases, correlating with CSF white blood cell counts.
Conclusions:
- Clinical diagnosis of EVM is difficult, highlighting the need for more effective diagnostic methods.
- The enhanced sensitivity and rapid turnaround time of RT-PCR offer significant clinical benefits for diagnosing EVM.
- RT-PCR represents a valuable advancement in the rapid and accurate detection of enteroviruses in CSF.
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