Diagnosis of meningococcal meningitis in Brazil by use of PCR

Abstract

Fever and a petechial rash are strongly associated with meningococcal disease in the city of Rio de Janeiro. Early antibiotic therapy is indicated and, consequently, a reduction of confirmed cases by culture, Gram stain, and latex agglutination test is expected. We evaluated a multiplex PCR assay to identify Neisseria meningitidis, Streptococcus pneumoniae and Haemophilus influenzae in biological samples from cases of non-culture proven meningitis with a petechial rash at presentation. To detect DNA in cerebrospinal fluid (n = 71) or blood (n = 5), a PCR screen was performed, based on the crgA, ply and bexA targets, respectively. Of the total, 70 CSF and 3 blood samples (96%) were positive by PCR for the presence of N. meningitidis DNA. Another PCR assay predicted in 82% of these samples N. meningitidis serogroups A (2%), B (60%), C (7%), X (3%), Y (2%), 29E (2%) or W135 (24%). In non-culture proven meningitis, PCR was found to be a valuable adjunct for the demonstration of meningococcal aetiology.

The utility of the polymerase chain reaction assay for aetiologic definition of unspecified bacterial meningitis cases

“…The results obtained with the PCR protocol described here revealed that cases of unspecified bacterial meningitis (G00.9, ISCD-10) were mostly caused by N. meningitidis (92%), compared with results published previously in which the patients (96%) had suspected meningococcal disease (Pedro et al 2007). This might be expected taking into account the prevalence of meningococcal disease and the high susceptibility of the bacteria to antibiotic clearance in the vasculature and CSF (Stephens 2007).…”

“…Bacterial DNA can be extracted from clinical samples of patients with clinically diagnosed meningitis, and amplified to detect the causative agent by this molecular approach (Clarke 2006). This enables confirmation of the disease-causing organism by the presence of its DNA, which makes it possible to disclose non-viable bacteria obtained from delayed lumbar punctures up to 72 h after commencement of parenteral antibiotics (Taha & Olcén 2004, Chanteau et al 2006, Pedro et al 2007). More recently, it has been used as a significant adjunct molecular laboratory technique for the demonstration of the most common aetiologies of bacterial meningitis in many Reference Centres (Taha et al 2005).…”

“…Despite the advent of glycoprotein conjugate vaccines and the development of new antimicrobial agents, bacterial infections of the central nervous system continue to pose a serious threat to the health and welfare of people world-wide (Finn 2004, Harrison 2006, particularly in developing nations (Greenwood 2000, Chanteau et al 2006, Pedro et al 2007). At least 1.2 million cases of bacterial meningitis are estimated to occur each year, and 135,000 of these patients die (WHO 1998).…”

“…At that time, laboratory techniques used in the isolation and identification of bacteria from the CSF of patients with clinical meningitis were implemented in the Infectious Diseases State Institute São Sebastião (IEISS), in the city of Rio de Janeiro. Conventional bacteriology has been used with poor results for case ascertainment during the last ten years, mainly due to the practice of administering antibiotics before admission and reluctance to performance lumbar punctures at presentation (Pedro et al 2007). The absence of an adjunct laboratory technique for detection of non-culturable bacteria has resulted in a great number of notifications of non-culture proven (unspecified) meningitis cases without a rash, with CSF white blood cell count compatible with pyogenic bacterial infection (Fig.…”

Neisseria meningitidis: a neglected cause of infectious haemorrhagic fever in the amazon rainforest

“…Many other conditions may cause acute fever with vasculitic rash and impaired consciousness into this country, like aetiologies rarely involved with this presentation -S. pneumoniae, Haemophilus influenzae, Staphylococcus aureus, and echovirus type 9 -or aetiologies presenting a later rash (4 th -6 th day) in the course of the disease -group Rickettsia species, dengue viruses, and other various viral haemorrhagic fevers -, many times with a different presentation and outcome [6,7,10,11,[21][22][23]. The most significant point to identify a meningococcal aetiology is a suggestive rash, which is a common feature of N. meningitidis invasive disease -meningitis and/or septicaemia -appearing on the first to second day of disease in 50 to 70% of reported cases [8,12,13,24,25]. Therefore, the early recognition of a petechial rash is fundamental to improve the outcome, as well as other features of acute meningitis, the most frequent presentation of meningococcal disease [13,21,24].…”