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CC BY-NC-ND 4.0 · Arq Neuropsiquiatr 2016; 74(10): 810-815
DOI: 10.1590/0004-282X20160119
ARTICLE

Red blood cells in cerebrospinal fluid as possible inhibitory factor for enterovirus RT-PCR

Presença de hemácias no líquido cefalorraquidiano como possível fator inibitório da RT-PCR para detecção de enterovirus
Sérgio Monteiro de Almeida
1   Universidade Federal do Paraná, Hospital de Clínicas, Seção de Virologia, Curitiba PR, Brasil;
2   Faculdades Pequeno Príncipe, Curitiba PR, Brasil;
3   Instituto de Pesquisa Pelé Pequeno Príncipe, Curitiba PR, Brasil.
,
Sçnia Mara Raboni
1   Universidade Federal do Paraná, Hospital de Clínicas, Seção de Virologia, Curitiba PR, Brasil;
,
Meri Bordignon Nogueira
1   Universidade Federal do Paraná, Hospital de Clínicas, Seção de Virologia, Curitiba PR, Brasil;
,
Luine R. Renaud Vidal
1   Universidade Federal do Paraná, Hospital de Clínicas, Seção de Virologia, Curitiba PR, Brasil;
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ABSTRACT

The presence of hemoglobin in samples are considered an important inhibitory factor for polymerase chain reaction (PCR). The aim of this study was to examine the influence of red blood cells (RBC)s in cerebrospinal fluid (CSF) as an inhibitory factor to reverse transcription polymerase chain reaction (RT-PCR) for enteroviruses (EV). Forty-four CSF samples from patients showing characteristics of viral meningitis were assessed for EV by RT-PCR. Viral RNA extracted with guanidine isothyocianate buffer and virus detection was performed by in-house nested PCR. Positivity for EV RT-PCR was higher in CSF samples without RBCs than in samples with RBCs: 13(26%) and 36(9.2%), p = 0.001. In the group with positive EV RT-PCR, the mean + SD CSF RBC was 37 ± 183 cell/mm3; the group with negative results had 580 + 2,890 cell/mm3 (p = 0.007). The acceptable upper limit for CSF RBCs that could not influence RT-PCR was 108 cells/mm3. CSF samples with negative results for EV RT-PCR have more erythrocytes.

RESUMO

A presença de hemoglobina em amostras de fluidos corporais é considerada um fator inibitório importante da reação em cadeia da polimerase (PCR). O objetivo deste estudo era examinar a influencia de hemácias no líquido cefalorraquidiano (LCR) como um fator inibitório da RT-PCR para enterovirus (EV). Quatrocentos e quarenta amostras de LCR de pacientes com características de meningite viral foram avaliados para enterovirus por RT-PCR. RNA viral foi extraído com tampão de isotiocianato de guanidina e a detecção viral foi feita com nested PCR in-house. A positividade do EV RT-PCR no LCR foi maior nas amostras de LCR sem hemácias do que as amostras com hemácias: 13 (26%) e 36 (9,2%), respectivamente (p = 0,001). No grupo com resultados EV RT-PCR positivo, a media ± DP do número de hemácias no LCR foi 37 ± 183 cell/mm3 e no grupo com resultados negativos foi 580 ± 2.890 cell/mm3 (p = 0,007). O limite superior aceitável de hemácias no LCR para não inibir o resultado do PCR foi 108 cells/mm3. As amostras de LCR com resultados negativos para RT-PCR EV tem mais eritrócitos em comparação com amostras com resultados positivos.

enterovirus - cerebrospinal fluid - reverse transcriptase polymerase chain reaction - hemoglobins - erythrocytes - meningitis
enterovirus - líquido cefalorraquidiano - reação em cadeia da polimerase via transcriptase reversa - hemoglobinas - eritrócitos - meningite


Publikationsverlauf

Eingereicht: 31. Dezember 2015

Angenommen: 14. Juli 2016

Artikel online veröffentlicht:
06. September 2023

© 2023. Academia Brasileira de Neurologia. This is an open access article published by Thieme under the terms of the Creative Commons Attribution-NonDerivative-NonCommercial License, permitting copying and reproduction so long as the original work is given appropriate credit. Contents may not be used for commecial purposes, or adapted, remixed, transformed or built upon. (https://creativecommons.org/licenses/by-nc-nd/4.0/)

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