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Polymerase Chain Reaction and Dot-Blot Hybridization for Leptospira Detection in Water Samples
Published on: June 14, 2024
Aseptic meningitis caused by Leptospira spp diagnosed by polymerase chain reaction
Eliete Caló Romero1, Roberta Morozetti Blanco, Paulo Hideki Yasuda
1Laboratório de Leptospirose, Seção de Bacteriologia, Divisão de Biologia Médica, Instituto Adolfo Lutz, São Paulo, SP, Brasil, 01246-902. ecromero@ial.sp.gov.br
Abstract:
Leptospirosis is a zoonotic disease caused by the pathogenic Leptospira spp. The clinical presentations are diverse, ranging from undifferentiated fever to fulminant disease including meningeal forms. The neurological leptospirosis forms are usually neglected. The aim of this study was to investigate leptospirosis as the cause of aseptic meningitis using different diagnostic techniques including the polymerase chain reaction (PCR). Thirty-nine cerebrospinal fluid (CSF) samples from patients presenting with meningeal abnormalities, predominance of lymphocytes and negative results by traditional microbiological tests were processed by leptospiral culture, anti-leptospiral antibody response and PCR. Leptospira spp DNA was detected in 23 (58.97%) of the CSF samples. Anti-leptospiral antibodies were found in 13 (33.33%) CSF samples. Twelve CSF samples were positive by PCR assay and negative by microscopic agglutination test (MAT) assay. Two CSF samples were positive by MAT and negative by PCR. The positive and negative agreement between both tests was 11 and 14, respectively. CSF samples from six cases of unknown diagnosis were positive by PCR assay. Eight cases showed positive results using PCR and MAT. Leptospirosis could be detected by PCR assay from the 3rd-26th day after illness onset. The sensitivity of the PCR was assessed with confirmed cases of leptospirosis (by MAT) and found to be 89.5%. All CSFs were negative by culture. PCR was found to be a powerful tool for diagnosing meningitis cases of leptospirosis. We recommend that it may be used as a supplementary diagnostic tool, especially in the early stages of the disease, when other diagnostic techniques such as serology are not sensitive.
Insights
Polymerase chain reaction (PCR) effectively detects Leptospira in aseptic meningitis cases, outperforming traditional methods. This powerful diagnostic tool aids early disease identification when serology is less sensitive.
Area of Science:
- Infectious Diseases
- Neurology
- Microbiology
Background:
- Leptospirosis, a zoonotic disease caused by Leptospira spp., presents with diverse symptoms, including neurological manifestations like meningitis.
- Neurological forms of leptospirosis are often overlooked, necessitating improved diagnostic approaches.
Purpose of the Study:
- To investigate the role of leptospirosis in aseptic meningitis using advanced diagnostic techniques.
- To evaluate the efficacy of polymerase chain reaction (PCR) in diagnosing neurological leptospirosis.
Main Methods:
- Analysis of 39 cerebrospinal fluid (CSF) samples from patients with aseptic meningitis.
- Utilized leptospiral culture, anti-leptospiral antibody detection (microscopic agglutination test - MAT), and PCR for diagnosis.
- Compared PCR results with traditional microbiological tests and serology.
Main Results:
- Leptospira spp. DNA was detected in 58.97% of CSF samples by PCR.
- PCR identified 12 cases missed by MAT, while MAT identified 2 cases missed by PCR.
- PCR demonstrated high sensitivity (89.5%) and could detect Leptospira from the 3rd to 26th day of illness.
Conclusions:
- PCR is a powerful and sensitive tool for diagnosing leptospirosis presenting as meningitis.
- PCR is particularly valuable in early disease stages when serological tests may be insensitive.
- The study recommends incorporating PCR as a supplementary diagnostic method for neurological leptospirosis.
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