Evaluation of a real-time PCR assay performance for the diagnosis of enteroviral meningitis
Vol. 75 (2016), BRIEF COMMUNICATION
Vol. 75 (2016)

Evaluation of a real-time PCR assay performance for the diagnosis of enteroviral meningitis

BRIEF COMMUNICATION
https://doi.org/10.53393/rial.2016.v75.33521
Published 2016-10-01
Bráulio Caetano Machado
Instituto Adolfo Lutz, Centro de Virologia, Núcleo de Doenças Entéricas, São Paulo, SP
Heloísa Rosa Vieira
Instituto Adolfo Lutz, Centro de Virologia, Núcleo de Doenças Entéricas, São Paulo, SP
Mayara Rhaissa de Moraes Alves
Instituto Adolfo Lutz, Centro de Virologia, Núcleo de Doenças Entéricas, São Paulo, SP
Rita de Cássia Compagnoli Carmona
Instituto Adolfo Lutz, Centro de Virologia, Núcleo de Doenças Entéricas, São Paulo, SP
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Keywords

enterovirus infections
meningitis
real time PCR

How to Cite

1.
Machado BC, Vieira HR, Alves MR de M, Carmona R de CC. Evaluation of a real-time PCR assay performance for the diagnosis of enteroviral meningitis. Rev Inst Adolfo Lutz [Internet]. 2016 Oct. 1 [cited 2024 Dec. 4];75:01-6. Available from: https://periodicos.saude.sp.gov.br/RIAL/article/view/33521
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Abstract

Enterovirus (EV) genus is the most frequent etiological agent causing viral meningitis worldwide. This study aimed at evaluating the performance of real-time reverse transcription-PCR (RT-qPCR) assay for detecting EV. A total of 616 cerebrospinal fluid (CSF) samples from patients with meningitis were selected, among those received at EV diagnosis laboratory from 1998 to 2013. RNAs were directly extracted from CSF by using QIAamp® Viral RNA Mini Kit, and TaqMan® RT-qPCR assay was applied. Evaluation was made by comparing the RT-qPCR results with those found in the cell culture for viral isolation method. Of 616 analyzed samples, 94 (15.20%) were positive for EV RNA on the RT-qPCR assay; and in the cell culture EV was isolated from 58 (9.40 %) samples. The assay showed sensitivity, specificity, positive predictive value and negative predictive value of 89.70 %, 92.40 %, 55.30 % and 98.90 %, respectively. In the present study, the RT-qPCR assay was slightly superior when compared to the viral culture technique for detecting EV from CSF samples. The TaqMan® RT-qPCR assay shows to be a fast and sensitive assay, easy to perform, and it shows a potential to improve the viral meningitis diagnosis in the public health laboratory in São Paulo State.

https://doi.org/10.53393/rial.2016.v75.33521
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References

1. Knowles NJ, Hovi T, Hyypiä T, King AM, Lindberg AM, Pallansch MA, et al. Picornaviridae. In: Virus Taxonomy: Classification and Nomenclature of Viruses: Ninth Report of the International Committee on Taxonomy of Viruses. San Diego: Elsevier; 2012.p.855-80.

2. Pallansch MA, Roos RP. Enteroviruses: polioviruses, coxsackieviruses, echoviruses, and newer enteroviruses. In: Knipe DM, Howley PM. 4ª ed. Philadelphia (PA): Lippincott Williams and Wilkins; 2001.p.723-75.

3. Abzug MJ. Presentation, diagnosis, and management of enterovirus infections in neonates. Paediatr Drugs. 2004;6(1):1-10. [DOI:10.2165/00148581-200406010-00001].

4. King RL, Lorch SA, Cohen DM, Hodinka RL, Cohn KA, Shah SS. Routine cerebrospinal fluid enterovirus polymerase chain reaction testing reduces hospitalization and antibiotic use for infants 90 days of age or younger. Pediatrics. 2007;120(3):489–96. [DOI:10.1542/peds. 2007-0252].

5. Romero JR. Reverse-transcription polymerase chain reaction detection of the enteroviruses. Arch Pathol Lab Med. 1999;123:1161-9.[DOI:10.1043/0003-998528-199929123<1161].

6. Nijhuis M, Van Maarseveen N, Schuurman R, Verkuijlen S, De Vos M, Hendriksen K, et al. Rapid and sensitive routine detection of all members of the genus enterovirus in different clinical specimens by real-time PCR. J Clin Microbiol. 2002;40(10):3666-70.[DOI:10.118/JCM-40.10.3666-3670.2002].

7. Pabbaraju K, Wong S, Wong AA, Tellier R. Detection of enteroviruses and parechoviruses by a multiplex real-time RT-PCR assay. Mol Cell Probes. 2015;29(2):81-5. [DOI:10.1016/j.mcp.2015.02.001].

8. Giulieri SG, Chapuis-Taillard C, Manuel O, Hugli O, Pinget C, Wasserfallen JB, et al. Rapid detection of enterovirus in cerebrospinal fluid by a fully-automated PCR assay is associated with improved management of aseptic meningitis in adult patients. J Clin Virol. 2015;62:58-62. [DOI:10.1016/j.jcv.2014.11.001].

9. Hierholzer JC, Killington RA. Virus isolation and quantification. In: Virology Methods Manual: Kangro HO, Mahy BWJ. London: Academic Press; 1996.p.24-45.

10. Verstrepen WA, Bruynseels P, Mertens AH. Evaluation of a rapid real-time RT-PCR assay for detection of enterovirus RNA in cerebrospinal fluid specimens. J Clin Virol. 2002;25:39-43. [DOI:10.1016/S1386-6532(02)00032-X].

11. Landis JR, Koch GG. The measurement of observer agreement for categorical data.Biometrics. 1977;33(1):159-74.[DOI:10.2307/2529310].

12. Oberste MS, Peñaranda S, Rogers SL, Henderson E, Nix WA. Comparative evaluation of Taq Man real time PCR and Semi nested VP1 PCR for detection of enteroviruses in clinical specimens. J Clin Virol. 2010;49(1):73-4. [DOI:10.1016/j.jcv.2010.06.022].

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