![]() Molecular tests for detecting nucleic acids of infectious agents in biological samples have been developed for C. neoformans penetrates the organism, lodging primarily in the lungs and later presents a notable tropism for the central nervous system 7,9,14. It is an important opportunist systemic mycosis that involves mainly immunosuppressed individuals and starts when C. In the last decades, cryptococcosis has been assuming a prominent role at public health level due to the growing number of AIDS individual cases 2,3,20. A análise comparativa dos três métodos, tinta da china, cultura e PCR, demonstrou que o último é muito mais sensível e específico, podendo ser aplicável como importante recurso laboratorial no diagnóstico da neurocriptococose.Ĭryptococcosis is a cosmopolitan infection caused by Cryptococcus neoformans, an encapsulated yeast pathogenic to humans and animals 1,13,18. A técnica de PCR pode detectar 1 célula/mL de líquor e é altamente específica. Os resultados demonstraram que a técnica tem alta sensibilidade, superior a cultura (85,7%) e ao teste da tinta da china (76,8%). Os pacientes eram portadores de meningite criptocócica (n = 56) e meningite ocasionada por outros agentes (n = 16). Foram estudados 72 líquors obtidos de casos de pacientes com e sem AIDS. A amplificação foi realizada nas áreas ITS e 5,6S do RNA ribossomal (rRNA). Keywords: Cryptococcus neoformans PCR Diagnostic methods AIDS CryptococcosisĪ detecção de Cryptococcus neoformans em líquor foi otimizada pela técnica de PCR. The comparative analysis of the methods showed that PCR is more sensitive and specific and is applicable as an important laboratorial resource for neurocryptococcosis diagnosis. PCR was found to be sensitive in detecting 1 cell/mL and highly specific since it did not amplify other fungal DNA. The results demonstrated that PCR test had the highest sensitivity rates, superior to culture (85.7%) and to India ink test (76.8%). ![]() ![]() The patients had cryptococcal meningitis (n = 56) and meningitis caused by other agents (n = 16). A total of 72 cerebrospinal fluid (CSF) samples were used, obtained from cases with and without AIDS. The amplification area was ITS and 5,6S which encodes the ribosomal RNA (rRNA). Tel: + 55 11 30917294, Fax: +55 11 3091 7354Ĭryptococcus neoformans detection was optimized using PCR technique with the objective of application in the clinical laboratory diagnosis. IIIDepartment of Microbiology, Institute of Biomedical Sciences, USP, São Paulo, SP, Brazil IIAdolfo Lutz Institute, São Paulo, SP, Brazil IUniversity of São Paulo, USP, Faculty of Pharmaceutical Sciences, Department of Clinical Analyses and Toxicology, São Paulo, SP, Brazil Regina Célia Paschoal I Mário Hiroyuki Hirata I Rosário Crespo Hirata I Márcia de Souza Carvalho Melhem II Amanda Latercia Tranches Dias III Claudete Rodrigues Paula III The application of mNGS, together with India ink staining, culture methods, and CrAg, may significantly improve the diagnostic precision in CM, thereby informing choice of appropriate antifungal treatment courses.Ĭerebrospinal fluid cryptococcal meningitis diagnosis next-generation sequencing pathogens.Neurocryptococcosis: diagnosis by PCR method MNGS is helpful for identifying Cryptococcus species. was detected concurrently in 33.33 % (4/12). The mNGS results of the CSF identified DNA sequences corresponding to Cryptococcus in 75 % of samples (9/12). India ink staining and culture of the CSF were positive for Cryptococcus in 83.33 % (10/12) of the samples 100 % (11/11) were positive via CrAg EIA. We aimed to assess the diagnostic performance of mNGS of cerebrospinal fluid (CSF) for prediction of cryptococcal meningitis (CM).Ī comparative evaluation of mNGS (performed on CSF samples) and conventional methods, including India ink staining, culture for fungi and cryptococcal-antigen (CrAg) detection by enzyme immunoassay, was performed on 12 consecutive non-HIV-infected patients with chronic or subacute CM. In recent years, metagenomic next-generation sequencing (mNGS) has become widely used in medical microbiology to detect pathogen infection.
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