Identification of Mycobacterium tuberculosis in the cerebrospinal fluid of patients with meningitis using nested PCR

Rios-Sarabia,Nora; Hernández-González,Olivia; González-Y-Merchand,Jorge; Gordillo,Guadalupe; Vázquez-Rosales,Guillermo; Muñoz-Pérez,Leopoldo; Torres,Javier; Maldonado-Bernal,Carmen

doi:10.3892/ijmm.2016.2698

Identification of Mycobacterium tuberculosis in the cerebrospinal fluid of patients with meningitis using nested PCR

Authors:
- Nora Rios-Sarabia
- Olivia Hernández-González
- Jorge González-Y-Merchand
- Guadalupe Gordillo
- Guillermo Vázquez-Rosales
- Leopoldo Muñoz-Pérez
- Javier Torres
- Carmen Maldonado-Bernal
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Affiliations: Medical Research Unit on Infectious Diseases, Hospital of Pediatrics, XXI Century National Medical Center, Mexican Social Security Institute, Mexico City 06720, Mexico, Laboratory of Molecular Microbiology, Department of Microbiology, National School of Biological Sciences, National Polytechnic Institute, Mexico City 11340, Mexico, Department of Infectious Diseases, Hospital of Pediatrics, XXI Century National Medical Center, Mexican Social Security Institute, Mexico City 06720, Mexico, Research Laboratory in Immunology and Proteomics, Children's Hospital of Mexico Federico Gomez, Mexico City 06720, Mexico
Published online on: August 4, 2016 https://doi.org/10.3892/ijmm.2016.2698
Pages: 1289-1295

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Abstract

Tuberculous meningitis (TBM) is the most severe form of tuberculosis. It is caused by Mycobacterium tuberculosis (M. tuberculosis; MT) and it is very difficult to diagnose. The symptoms are similar to other infectious neurological diseases, such as neurocysticercosis, neuroborreliosis, or herpes viral infection. The aim of this study was to identify tuberculosis (TB) in cases of meningitis with clinical and laboratory evidence suggestive of TBM, and to confirm our findings with molecular tests for TB infection. We recruited patients with neurological symptoms who were examined at the neurology services of Hospitals of Instituto Mexicano del Seguro Social (IMSS) in Mexico City. A total of 144 consecutive patients with suggestive infectious meningitis were initially included; 94 cases of meningitis with clinical and laboratory evidence suggestive of TBM were included, but only 50 of these cases fulfilled the criteria for probable TBM. As the controls, we included 50 cases of meningitis with clinical and laboratory evidence suggestive of non-TBM. Cerebrospinal fluid (CSF) was collected from all 100 patients (cases and controls) and tested for TB by multiplex and nested PCR analyses. Nested PCR detected 0.1 fg of M. tuberculosis DNA. TB infection was confirmed with molecular tests in 49 patients from the 50 cases suggestive of TBM and in 1 of the 50 non-TBM cases. The analysis exhibited a sensitivity of 98.0%, a specificity of 92.0%, a positive predictive value of 88.0% and a negative predictive value of 98.0%. The use CSF for the analyses proved to be effective for the rapid diagnosis of TBM using a developed system of multiplex and nested PCR analyses in patients presenting neurological symptoms.

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