![]() Group cryptococcal meningitis and AIDS (CM/AIDS) consisted of 96 CSF samples from patients with CM/AIDS. The clinical samples were divided into two groups. The assays were evaluated analyzing 165 CSF samples with commonly used, well-known laboratory and clinical diagnoses. The sensitivity and specificity of the methodology were evaluated using Cryptococcus reference strains in the first part and CSF samples from patients with neurological diseases in the second part of the study. The aim of this study is to evaluate the use of PCR for molecular diagnosis of cryptococcal meningitis. Hence, molecular methods such as PCR have proven to be useful for this purpose, since they can provide a definitive diagnosis as well as evaluation of specific treatment. Molecular methods such as polymerase chain reaction (PCR) potentially identify the fungus and the genotype of the strain.Ĭonsidering that no method is entirely effective in determining the presence of this pathogen, it is helpful to diagnose on the basis of results obtained from different methodologies to rule out false-negative or false-positive results. 11–15 Nevertheless, there is scarce information about the use of molecular diagnosis in clinical samples. Several studies have demonstrated the use of molecular methods for determining the genetic characteristics of Cryptococcus species. 10 Routinely, microscopic examination, cryptococcal polysaccharide antigen, and culture of cerebrospinal fluid (CSF) samples lead to the diagnosis. 7–9 A rapid laboratorial diagnosis is extremely important to identify Cryptococcus species. Patients with cryptococcal meningitis usually present severe neurological manifestations, particularly secondary to intracranial hypertension. 4,7 However, until today, cryptococcal meningitis remains a common complication in Brazil and other Latin American countries as it represents the primary cause of opportunistic meningitis and the second-most frequent neurologic opportunistic infection in HIV-infected patients. 6 Fortunately, in recent years, AIDS-associated cryptococcal meningitis has decreased dramatically in Brazil, which is a middle-income country with universal access to antiretroviral therapy. 5 Latin America, as a global region, has the third highest incidence of the disease, with around 54,400 cases annually. 1–4Ĭryptococcal meningitis affects approximately one million persons in the world each year and results in more than 400,000 deaths within three months after the onset of the disease. The course of infection is often a life-threatening disease that is typically observed in later stages of acquired immunodeficiency syndrome (AIDS). gattii species complex in cerebrospinal fluid samples from patients with clinical suspicion of cryptococcal meningitis.Ĭryptococcal meningitis is one of the most serious systemic mycoses, mainly affecting immunocompromised patients and causing high morbidity and mortality. These data suggest that the CN4/CN5 primer set was highly sensitive for the identification of C. The specificity based on the negative samples from the CTL/AIDS group was 98.5% in both primer sets. This primer set was negative for two standard strains. ![]() neoformans were observed only in 64.6% of the cryptococcal meningitis and AIDS group. gattii was observed in the clinical samples of either group. With the multiplex, no 448-bp product of C. ResultsĬN4–CN5 primer set was positive in all Cryptococcus standard strains and in 94.8% in DNA samples from cryptococcal meningitis and AIDS group. Two primer sets were tested (CN4–CN5 and the multiplex CNa70S–CNa70A/CNb49S–CNb-49A that amplify a specific product for C. gattii species complex reference strains and 165 cerebrospinal fluid samples from patients with neurological diseases divided into two groups: 96 patients with cryptococcal meningitis and AIDS and 69 patients with other neurological opportunistic diseases (CRL/AIDS). The sensitivity and specificity of the methodology were evaluated using eight Cryptococcus neoformans/C. This study evaluated the use of polymerase chain reaction for cryptococcal meningitis diagnosis in clinical samples.
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